Method for predicting risk of aggressive or recurrent cancer

Abstract

A method of predicting the risk of invasive or recurrent prostate cancer in a subject, the method comprising the step of determining the expression of at least two genes selected from the group consisting of TTC21B, MCM3, MTUS1, FOXM1, CUL4A, GPC1, PCNA, IRF7, BRIP1, ULK1 and SPAG5, or a protein encoded by said genes, in a biological sample from the subject, wherein when a combination of normalized expression values of the at least two genes or a protein encoded by said genes is altered relative to a combination of normalized expression reference values of the at least two genes or by said encoded protein in a biological sample from a subject suffering from non-invasive or non-recurrent prostate cancer, the risk of predicting the individual to have aggressive or recurrent prostate cancer is increased.

Description

Technical Field

[0001] This invention relates to a method for predicting the risk of an aggressive or recurrent cancer in a subject. Specifically, this invention relates to a method for predicting the risk of an aggressive cancer and the risk of cancer recurrence in a subject. Background Technology

[0002] Thanks to increased awareness of prostate cancer, widespread prostate-specific antigen (PSA) screening, and digital rectal examination (DRE) testing, many prostate cancers (approximately 60%) are detected at an early stage. The majority (60%) are detected in men over 65 years of age. About 80% of patients have slow-growing or indolent prostate cancer and may go untreated while undergoing frequent surveillance known as active surveillance. The remaining 20% ​​have invasive prostate cancer, which has the potential to metastasize to other parts of the body and requires aggressive treatment. Historically, most prostate cancer patients have received aggressive treatment, with many undergoing radical prostatectomy combined with radiation therapy, hormone therapy, or chemotherapy, resulting in significant lifelong adverse side effects. This unnecessary overtreatment significantly increases costs for both patients and the healthcare system. Current clinical tools cannot determine the true risk of metastasis; therefore, accurately identifying patients with invasive prostate cancer (i.e., predicting metastatic potential) represents a major unmet clinical need and is crucial for selecting appropriate treatment options for prostate cancer patients.

[0003] Prostate cancer is the second most common cancer in men after non-melanoma skin cancer (accounting for 26% of all male cancer diagnoses in the UK), the second leading cause of cancer death in men after lung cancer, and the fourth most prevalent cancer globally. An estimated 1.2 million men worldwide are diagnosed with prostate cancer each year. The average age at diagnosis is 67 years, with approximately 25% of cases occurring in men under 65. Due to increased PSA screening, most patients are diagnosed with early-stage disease.

[0004] A major clinical challenge for clinicians and patients is how best to treat prostate cancer. In cases of indolent cancer, or if a patient requires radical treatment for an aggressive cancer, it must be determined whether active surveillance is the appropriate clinical approach. Radical prostatectomy and radiation therapy expose patients to significant and lifelong negative side effects and quality-of-life problems, including incontinence, erectile dysfunction, mood disturbances, and depression. Statistically, 80% of men will develop indolent, slow-growing, low-grade prostate cancer, which can be safely managed through active surveillance, while 20% of these men will require invasive treatment. To date, approximately 76% of men are treated for invasive disease, leading to unnecessary overtreatment and exposing patients to significant side effects. Accurately determining the grade, stage, and invasiveness of prostate cancer is crucial for appropriate patient treatment stratification. Increasing the number of men under active surveillance will (i) reduce overtreatment and improve the ability to treat high-risk diseases quickly, (ii) reduce the number of men who unnecessarily receive curative treatment and experience adverse effects, and (iii) reduce the cost of treating and managing these adverse effects.

[0005] A group of biomarkers for determining the presence of cancer in a subject is known in the art. For example, WO2015 / 087088 describes a method for predicting the risk of invasive or recurrent prostate cancer, where MTUS1 is one of many biomarkers. WO2016 / 042164 describes a method for predicting the risk of cancer recurrence in an individual with cancer, comprising the step of determining the positive expression of at least two genes or proteins encoded by those genes in a cancer sample from the individual, the genes being selected from the group consisting of: FOXM1, UHRF1, PTTG1, E2F1, MYBL2, HMGB2, ATAD2, E2F8, ZNF367, and TCF19. WO03 / 078662 describes a method for measuring mRNA levels in biopsied tumor tissue for the diagnosis and treatment of breast cancer. LV Daojun et al. (Cancer Immunology Immunotherapy, Vol. 70, pp. 3587-3602 (2021)) describe the use of immune-related gene-based prognostic markers (IRGPS) incorporating the biomarker TIGIT for identifying high-risk prostate cancer subjects. Abou-ouf, Hatem et al. (Journal of Cancer Research and Clinical Oncology, Vol. 144, pp. 883-891 (2018)) describe 10-gene markers in prostate cancer that can be used to differentiate between aggressive and indolent disease within low-risk and intermediate-risk patient groups. WO2012 / 135397 describes a method for discovering biomarkers (biomarkers) for any cancer whose growth may be regulated by mitochondrial oxidative metabolism. WO2020 / 214718 describes a method for identifying cancers associated with poor prognosis. WO2015 / 127101 describes a method for diagnosing invasive prostate cancer, which includes determining the expression level of each of the genes encoding the forkhead box protein M1 (FOXM1) and the centromere protein F (CENPF) in a test sample.

[0006] The object of this invention is to overcome at least one of the problems mentioned above. Summary of the Invention

[0007] Prostate cancer is the second most common cancer in men after non-melanoma skin cancer, the second leading cause of cancer death in men after lung cancer, and the fourth most prevalent cancer globally. Approximately 80% of prostate cancer patients have slow-growing cancer and can safely remain untreated while undergoing active monitoring; while 20% of prostate cancer patients have aggressive cancer with high metastatic potential and require aggressive treatment involving radical prostatectomy (RP) combined with radiation therapy, hormone therapy, or chemotherapy.

[0008] Current clinical tools cannot accurately identify whether a patient has invasive prostate cancer, thus overtreatment is a major problem in the clinical management of prostate cancer, as many patients undergo radical prostatectomy, leading to serious complications including infection, incontinence, erectile dysfunction, and depression. To differentiate between patients requiring invasive treatment and those who can safely undergo active surveillance, accurate prognostic tests are needed to determine a patient's risk of prostate cancer recurrence; this clinical need remains unmet. The development of such prognostic tests will reduce overtreatment of prostate cancer patients, thereby sparing many individuals from the numerous adverse side effects of treatment.

[0009] The method of the claimed invention is based on measuring changes in gene expression levels that are highly correlated with cancer progression and invasiveness. This test measures, for example, the invasiveness of prostate cancer and its potential for recurrence, thereby enabling more personalized treatment strategies.

[0010] This method is a multigene prognostic marker used to assess the probability of aggressive disease in men, for example, those recently diagnosed with prostate cancer. RNA samples extracted from formalin-fixed, paraffin-embedded (FFPE) prostate tumor tissue were tested using RT-qPCR. Those deemed to be in a lower-risk category (i.e., with a lower risk score) through testing can safely forgo immediate definitive treatment, while those deemed to be in a higher-risk category (i.e., with a higher risk score) may be considered for intensive treatment. Low risk refers to a lower risk of aggressive or recurrent disease, and high risk refers to a higher risk of aggressive or recurrent disease.

[0011] This test employs a patient risk score generated by summing the expression levels of at least two genes and comparing it to a reference risk score. The expression level of each gene is typically normalized. In some embodiments, a function of the expression levels of each gene is summed and compared to a reference risk score. Examples are described in more detail below.

[0012] This method may include determining the expression of at least two genes or a combination of proteins encoded by said genes from a sample from a subject. The expression level of each of the at least two genes is normalized against a reference gene (e.g., a set of one or more housekeeping genes). The normalized expression levels or a function of the normalized expression levels (e.g., weighted normalized expression levels, where the weights are gene-specific) are then summed to provide a test risk score. The same exercise is performed on control samples (e.g., from a patient with non-invasive cancer or from a subject without recurrence) to provide a reference risk score. When two results / scores are obtained, they are compared to each other. When the normalized expression of the combination of genes from the test sample changes (increases or decreases) relative to the normalized expression of the combination of genes in the control sample, the test subject has a higher risk of invasive or recurrent disease. The combination is the sum of the gene-specific weighted normalized expressions of the genes.

[0013] A method is provided for predicting the risk of an individual for invasive or recurrent cancer, the method comprising: determining the expression of at least two genes or proteins encoded by said genes in a biological sample obtained from the individual, the genes being selected from TTC21B, MCM3, MTUS1, FOXM1, CUL4A, GPC1, PCNA, IRF7, BRIP1, ULK1, and SPAG5, wherein an increased risk of the individual for invasive or recurrent cancer is predicted when the combination of normalized expression values ​​of said at least two genes or proteins encoded by said genes changes relative to the combination of normalized expression values ​​of said at least two genes or proteins encoded by said genes in a biological sample from a subject with non-invasive or non-recurrent cancer.

[0014] Expression values ​​can be, for example, copy number or a function of copy number, such as gene-specific weighted copy number. Gene-specific weights can be determined based on the relative contribution of a specific gene to the risk of an aggressive or recurrent phenotype, such as by analyzing gene levels in a cohort of cancer patients including those with good and poor outcomes. Examples of such patient cohorts are described in the TCGA cohorts under the heading "Statistical Analysis" in the Materials and Methods section.

[0015] A method for predicting the risk of cancer recurrence or progression in an individual with cancer after treatment, the method comprising: determining the expression of at least two genes or proteins encoded by said genes in a biological sample obtained from the individual, the genes being selected from TTC21B, MCM3, MTUS1, FOXM1, CUL4A, GPC1, PCNA, IRF7, BRIP1, ULK1, and SPAG5, wherein an increase in the risk of cancer recurrence or progression in the individual after treatment is predicted when the combination of normalized expression values ​​of said at least two genes or proteins encoded by said genes changes relative to the combination of normalized expression values ​​of said at least two genes from a biological sample obtained from a subject who has not experienced cancer recurrence or progression after treatment.

[0016] On the one hand, the treatment is selected from active monitoring, surgery and / or radiation therapy, neoadjuvant therapy, adjuvant systemic therapy, or a combination thereof.

[0017] A method for determining whether an individual's cancer is an aggressive type of cancer, the method comprising: measuring the expression of at least two genes or proteins encoded by said genes in a biological sample from the individual, the genes being selected from TTC21B, MCM3, MTUS1, FOXM1, CUL4A, GPC1, PCNA, IRF7, BRIP1, ULK1, and SPAG5, wherein an increased risk of the individual having an aggressive type of cancer is predicted when the combination of normalized expression values ​​of said at least two genes or proteins encoded by said genes changes relative to the combination of normalized gene expression levels of said at least two genes from a subject who does not have aggressive cancer.

[0018] A method for determining the 5-year or 10-year survival rate of an individual diagnosed with cancer, the method comprising: measuring the expression of at least two genes or proteins encoded by those genes in a biological sample from the individual, the genes being selected from TTC21B, MCM3, MTUS1, FOXM1, CUL4A, GPC1, PCNA, IRF7, BRIP1, ULK1, and SPAG5, wherein a decrease in the individual's 5-year or 10-year survival rate is predicted when the combination of normalized expression values ​​of the at least two genes or proteins encoded by said genes changes relative to the combination of normalized expression values ​​of the at least two genes from a biological sample obtained from a subject who survived the cancer 5 or 10 years after the cancer diagnosis.

[0019] A method for determining the 5-year or 10-year survival rate of an individual diagnosed with prostate cancer, the method comprising: measuring the expression of at least two genes or proteins encoded by those genes in a biological sample from the individual, the genes being selected from TTC21B, MCM3, MTUS1, FOXM1, CUL4A, GPC1, PCNA, IRF7, BRIP1, ULK1, and SPAG5, wherein a decrease in the individual's 5-year or 10-year survival rate is predicted when the combination of normalized expression values ​​of the at least two genes or proteins encoded by said genes changes relative to the combination of normalized expression values ​​of the at least two genes from a biological sample obtained from a subject who has survived the prostate cancer 5 or 10 years after the cancer diagnosis.

[0020] A method for identifying an individual suitable for treatment with a therapy for preventing cancer recurrence or progression, the method comprising: measuring the expression of at least two genes or proteins encoded by these genes in a biological sample from the individual, the genes being selected from TTC21B, MCM3, MTUS1, FOXM1, CUL4A, GPC1, PCNA, IRF7, BRIP1, ULK1, and SPAG5, wherein the individual is predicted to be unsuitable for the treatment if the combination of normalized expression values ​​of the at least two genes or proteins encoded by said genes is the same as or increases after treatment with the therapy compared to the combination of normalized expression values ​​of the at least two genes in a biological sample from the same individual before treatment with the therapy. A different treatment should be considered and offered to the individual, and another biological sample from the individual should be measured after the different treatment is administered in the same manner as described above to determine whether the different treatment is appropriate. If, after treatment, the combination of normalized expression values ​​of at least two genes or proteins encoded by said genes decreases relative to the combination of normalized expression values ​​of the at least two genes in a biological sample from the same individual before treatment, the individual is predicted to be suitable for the treatment.

[0021] A method for monitoring the effectiveness of cancer treatment in an individual with cancer, the method comprising: determining the expression of at least two genes or proteins encoded by these genes in a biological sample from the individual, the genes being selected from TTC21B, MCM3, MTUS1, FOXM1, CUL4A, GPC1, PCNA, IRF7, BRIP1, ULK1, and SPAG5, wherein the treatment is ineffective and an adverse outcome is predicted for the individual when the combination of normalized expression values ​​of the at least two genes or proteins encoded by said genes is the same as or increases after administration of the treatment relative to the combination of normalized expression values ​​of the at least two genes in a biological sample from the same individual before administration of the treatment. In this case, the treatment regimen can be discontinued, and a new treatment regimen can be administered to the treated individual with cancer, or treatment can be discontinued entirely. If, after treatment, the combination of normalized expression values ​​of at least two genes or proteins encoded by said genes decreases relative to the combination of normalized expression values ​​of at least two genes in a biological sample from the same individual before treatment, the treatment is effective and the individual is predicted to have improved outcomes.

[0022] A method for predicting an individual's risk of invasive or recurrent cancer, the method comprising the step of determining the expression of at least one gene or a protein encoded by said gene in a biological sample obtained from the individual, said gene being selected from TTC21B, FOXM1, PCNA, IRF7, ULK1, and SPAG5, wherein an increase in the risk of the individual having invasive or recurrent cancer is predicted when the normalized expression value of said at least one gene or the protein encoded by said gene changes compared to a normalized expression value from a biological sample from a subject whose cancer is non-invasive or indolent.

[0023] A method for predicting an individual's risk of invasive or recurrent cancer, the method comprising the step of determining the expression of at least one gene or a protein encoded by said gene in a biological sample obtained from the individual, said gene being selected from MCM3, MTUS1, GPC1, BRIP1, and CUL4A, wherein an increase in the risk of the individual having invasive or recurrent cancer is predicted when the normalized expression value of said at least one gene or the protein encoded by said gene changes compared to a normalized expression value from a biological sample from a subject whose cancer is non-invasive or indolent.

[0024] A method for predicting the risk of an individual for invasive or recurrent cancer, the method comprising: determining the expression of at least two genes or proteins encoded by said genes in a biological sample obtained from the individual, the genes being selected from TTC21B, FOXM1, PCNA, IRF7, ULK1, and SPAG5, wherein an increased risk of the individual for invasive or recurrent cancer is predicted when the combination of normalized expression values ​​of said at least two genes or proteins encoded by said genes changes relative to the combination of normalized expression values ​​of said at least two genes from a subject with non-invasive or non-recurrent cancer.

[0025] A method for predicting an individual's risk of invasive or recurrent cancer, the method comprising measuring the expression of at least two genes or proteins encoded by said genes in a biological sample obtained from the individual, the genes being selected from MCM3, MTUS1, BRIP1, CUL4A, and GPC1, wherein an increased risk of invasive or recurrent cancer is predicted when the combination of normalized expression values ​​of the at least two genes or proteins encoded by said genes changes relative to the combination of normalized expression values ​​of the at least two genes from a subject with non-invasive or non-recurrent cancer.

[0026] A method for predicting an individual's risk of invasive or recurrent cancer, the method comprising the step of determining the expression of the TTC21B gene or a protein encoded by said gene in a biological sample obtained from the individual, wherein an increased risk of the individual having invasive or recurrent cancer is predicted when the normalized expression value of the TTC21B gene or the protein encoded by said gene changes relative to the normalized expression value of the TTC21B gene in a sample from an individual whose cancer is non-invasive or indolent.

[0027] A method for predicting an individual's risk of invasive or recurrent cancer, the method comprising the step of determining the expression of the MTUS1 gene or a protein encoded by said gene in a biological sample obtained from the individual, wherein an increased risk of the individual having invasive or recurrent cancer is predicted when the normalized expression value of the MTUS1 gene or the protein encoded by said gene changes relative to the normalized expression value of the MTUS1 gene in a sample from an individual with cancer.

[0028] In one embodiment, at least one gene is CUL4A. In one embodiment, at least one gene is TTC21B. In one embodiment, at least one gene is MCM3. In one embodiment, at least one gene is GPC1. In one embodiment, at least one gene is PCNA. In one embodiment, at least one gene is MTUS1. In one embodiment, at least one gene is IRF7. In one embodiment, at least one gene is BRIP1. In one embodiment, at least one gene is ULK1. In one embodiment, at least one gene is SPAG5. In one embodiment, at least one gene is FOXM1.

[0029] In one embodiment, the selected at least two genes are CUL4A and TTC21B. In one embodiment, the selected at least two genes are CUL4A and MCM3. In one embodiment, the selected at least two genes are CUL4A and GPC1. In one embodiment, the selected at least two genes are CUL4A and PCNA. In one embodiment, the selected at least two genes are CUL4A and MTUS1. In one embodiment, the selected at least two genes are CUL4A and IRF7. In one embodiment, the selected at least two genes are CUL4A and BRIP1. In one embodiment, the selected at least two genes are CUL4A and ULK1. In one embodiment, the selected at least two genes are CUL4A and SPAG5. In one embodiment, the selected at least two genes are TTC21B and MCM3. In one embodiment, the selected at least two genes are TTC21B and GPC1. In one embodiment, the selected at least two genes are TTC21B and PCNA. In one embodiment, the selected at least two genes are TTC21B and MUTS1. In one embodiment, the selected at least two genes are TTC21B and IRF7. In one embodiment, the selected at least two genes are TTC21B and BRIP1. In one embodiment, the selected at least two genes are TTC21B and ULK1. In one embodiment, the selected at least two genes are TTC21B and SPAG5. In one embodiment, the selected at least two genes are MCM3 and GPC1. In one embodiment, the selected at least two genes are MCM3 and PCNA. In one embodiment, the selected at least two genes are MCM3 and MTUS1. In one embodiment, the selected at least two genes are MCM3 and IRF7. In one embodiment, the selected at least two genes are MCM3 and BRIP1. In one embodiment, the selected at least two genes are MCM3 and UKL1. In one embodiment, the selected at least two genes are MCM3 and SPAG5. In one embodiment, the selected at least two genes are GPC1 and PCNA. In one embodiment, the selected at least two genes are GPC1 and MTUS1. In one embodiment, the selected at least two genes are GPC1 and IRF7. In one embodiment, the selected at least two genes are GPC1 and BRIP1. In one embodiment, the selected at least two genes are GPC1 and ULK1. In one embodiment, the selected at least two genes are GPC1 and SPAG5. In one embodiment, the selected at least two genes are PCNA and MTUS1. In one embodiment, the selected at least two genes are PCNA and IRF7. In one embodiment, the selected at least two genes are PCNA and BRIP1.In one embodiment, the selected at least two genes are PCNA and ULK1. In one embodiment, the selected at least two genes are PCNA and SPAG5. In one embodiment, the selected at least two genes are MTUS1 and IRF7. In one embodiment, the selected at least two genes are MTUS1 and BRIP1. In one embodiment, the selected at least two genes are MTUS1 and ULK1. In one embodiment, the selected at least two genes are MTUS1 and SPAG5. In one embodiment, the selected at least two genes are IRF7 and BRIP1. In one embodiment, the selected at least two genes are IRF7 and ULK1. In one embodiment, the selected at least two genes are IRF7 and SPAG5. In one embodiment, the selected at least two genes are BRIP1 and ULK1. In one embodiment, the selected at least two genes are BRIP1 and SPAG5. In one embodiment, the selected at least two genes are ULK1 and SPAG5. In one embodiment, the selected at least two genes are CUL4A and FOXM1. In one embodiment, the selected at least two genes are FOXM1 and TTC21B. In one embodiment, the selected at least two genes are FOXM1 and MCM3. In one embodiment, the selected at least two genes are FOXM1 and GPC1. In one embodiment, the selected at least two genes are FOXM1 and PCNA. In one embodiment, the selected at least two genes are FOXM1 and MTUS1. In one embodiment, the selected at least two genes are FOXM1 and IRF7. In one embodiment, the selected at least two genes are FOXM1 and BRIP1. In one embodiment, the selected at least two genes are FOXM1 and ULK1. In one embodiment, the selected at least two genes are FOXM1 and SPAG5.

[0030] In one embodiment, at least three genes are selected, and the selected genes are CUL4A, TTC21B, and MCM3. In one embodiment, the selected at least three genes are CUL4A, TTC21B, and GPC1. In one embodiment, the selected at least three genes are CUL4A, TTC21B, and PCNA. In one embodiment, the selected at least three genes are CUL4A, TTC21B, and MTUS1. In one embodiment, the selected at least three genes are CUL4A, TTC21B, and IRF7. In one embodiment, the selected at least three genes are CUL4A, TTC21B, and BRIP1. In one embodiment, the selected at least three genes are CUL4A, TTC21B, and ULK1. In one embodiment, the selected at least three genes are CUL4A, TTC21B, and SPAG5. In one embodiment, the selected at least three genes are TTC21B, MCM3, and GPC1. In one embodiment, the selected at least three genes are TTC21B, MCM3, and PCNA. In one embodiment, the selected at least three genes are TTC21B, MCM3, and MTUS1. In one embodiment, the selected at least three genes are TTC21B, MCM3, and IRF7. In one embodiment, the selected at least three genes are TTC21B, MCM3, and BRIP1. In one embodiment, the selected at least three genes are TTC21B, MCM3, and ULK1. In one embodiment, the selected at least three genes are TTC21B, MCM3, and SPAG5. In one embodiment, the selected at least three genes are MCM3, GPC1, and PCNA. In one embodiment, the selected at least three genes are MCM3, GPC1, and MTUS1. In one embodiment, the selected at least three genes are MCM3, GPC1, and IRF7. In one embodiment, the selected at least three genes are MCM3, GPC1, and BRIP1. In one embodiment, the selected at least three genes are MCM3, GPC1, and ULK1. In one embodiment, the selected at least three genes are MCM3, GPC1, and SPAG5. In one embodiment, the selected at least three genes are GPC1, PCNA, and MTUS1. In another embodiment, the selected at least three genes are GPC1, PCNA, and MTUS1. In yet another embodiment, the selected at least three genes are GPC1, PCNA, and IRF7. In yet another embodiment, the selected at least three genes are GPC1, PCNA, and BRIP1. In yet another embodiment, the selected at least three genes are GPC1, PCNA, and ULK1. In yet another embodiment, the selected at least three genes are GPC1, PCNA, and SPAG5.In one embodiment, the selected at least three genes are PCNA, MTUS1, and IRF7. In one embodiment, the selected at least three genes are PCNA, MTUS1, and BRIP1. In one embodiment, the selected at least three genes are PCNA, MTUS1, and ULK1. In one embodiment, the selected at least three genes are PCNA, MTUS1, and SPAG5. In one embodiment, the selected at least three genes are MTUS1, IRF7, and BRIP1. In one embodiment, the selected at least three genes are MTUS1, IRF7, and ULK1. In one embodiment, the selected at least three genes are MTUS1, IRF7, and SPAG5. In one embodiment, the selected at least three genes are IRF7, BRIP1, and ULK1. In one embodiment, the selected at least three genes are IRF7, BRIP1, and SPAG5. In one embodiment, the selected at least three genes are BRIP1, ULK1, and SPAG5. In one embodiment, the selected at least three genes are CUL4, MCM3, and GPC1. In one embodiment, the selected at least three genes are CUL4, MCM3, and PCNA. In one embodiment, the selected at least three genes are CUL4, MCM3, and MTUS1. In one embodiment, the selected at least three genes are CUL4, MCM3, and IRF7. In one embodiment, the selected at least three genes are CUL4, MCM3, and BRIP1. In one embodiment, the selected at least three genes are CUL4, MCM3, and ULK1. In one embodiment, the selected at least three genes are CUL4, MCM3, and SPAG5. In one embodiment, the selected at least three genes are CUL4, GPC1, and PCNA. In one embodiment, the selected at least three genes are CUL4, GPC1, and MTUS1. In one embodiment, the selected at least three genes are CUL4, GPC1, and IRF7. In one embodiment, the selected at least three genes are CUL4, GPC1, and BRIP1. In one embodiment, the selected at least three genes are CUL4, GPC1, and ULK1. In one embodiment, the selected at least three genes are CUL4, GPC1, and SPAG5. In one embodiment, the selected at least three genes are CUL4, PCNA, and MTUS1. In one embodiment, the selected at least three genes are CUL4, PCNA, and IRF7. In one embodiment, the selected at least three genes are CUL4, PCNA, and BRIP1. In one embodiment, the selected at least three genes are CUL4, PCNA, and ULK1. In one embodiment, the selected at least three genes are CUL4, PCNA, and SPAG5.In one embodiment, the selected at least three genes are CUL4, MTUS1, and IRF7. In one embodiment, the selected at least three genes are CUL4, MTUS1, and BRIP1. In one embodiment, the selected at least three genes are CUL4, MTUS1, and ULK1. In one embodiment, the selected at least three genes are CUL4, MTUS1, and SPAG5. In one embodiment, the selected at least three genes are CUL4, IRF7, and BRIP1. In one embodiment, the selected at least three genes are CUL4, IRF7, and ULK1. In one embodiment, the selected at least three genes are CUL4, IRF7, and SPAG5. In one embodiment, the selected at least three genes are CUL4, BRIP1, and ULK1. In one embodiment, the selected at least three genes are CUL4, BRIP1, and SPAG5. In one embodiment, the selected at least three genes are CUL4, ULK1, and SPAG5. In one embodiment, the selected at least three genes are TTC21B, GPC1, and PCNA. In one embodiment, the selected at least three genes are TTC21B, GPC1, and PCNA. In one embodiment, the selected at least three genes are TTC21B, GPC1, and MTUS1. In one embodiment, the selected at least three genes are TTC21B, GPC1, and IRF7. In one embodiment, the selected at least three genes are TTC21B, GPC1, and BRIP1. In one embodiment, the selected at least three genes are TTC21B, GPC1, and ULK1. In one embodiment, the selected at least three genes are TTC21B, GPC1, and SPAG5. In one embodiment, the selected at least three genes are TTC21B, PCNA, and MTUS1. In one embodiment, the selected at least three genes are TTC21B, PCNA, and IRF7. In one embodiment, the selected at least three genes are TTC21B, PCNA, and BRIP1. In one embodiment, the selected at least three genes are TTC21B, PCNA, and ULK1. In one embodiment, the selected at least three genes are TTC21B, PCNA, and SPAG5. In one embodiment, the selected at least three genes are TTC21B, MTUS1, and IRF7. In one embodiment, the selected at least three genes are TTC21B, MTUS1, and BRIP1. In one embodiment, the selected at least three genes are TTC21B, MTUS1, and ULK1. In one embodiment, the selected at least three genes are TTC21B, MTUS1, and SPAG5. In one embodiment, the selected at least three genes are TTC21B, IRF7, and BRIP1.In one embodiment, the selected at least three genes are TTC21B, IRF7, and ULK1. In one embodiment, the selected at least three genes are TTC21B, IRF7, and SPAG5. In one embodiment, the selected at least three genes are TTC21B, BRIP1, and ULK1. In one embodiment, the selected at least three genes are TTC21B, BRIP1, and SPAG5. In one embodiment, the selected at least three genes are TTC21B, ULK1, and SPAG5. In one embodiment, the selected at least three genes are MCM3, PCNA, and MTUS1. In one embodiment, the selected at least three genes are MCM3, PCNA, and IRF7. In one embodiment, the selected at least three genes are MCM3, PCNA, and BRIP1. In one embodiment, the selected at least three genes are MCM3, PCNA, and ULK1. In one embodiment, the selected at least three genes are MCM3, PCNA, and SPAG5. In one embodiment, the selected at least three genes are MCM3, MTUS1, and IRF7. In one embodiment, the selected at least three genes are MCM3, MTUS1, and BRIP1. In one embodiment, the selected at least three genes are MCM3, MTUS1, and ULK1. In one embodiment, the selected at least three genes are MCM3, MTUS1, and SPAG5. In one embodiment, the selected at least three genes are MCM3, IRF7, and BRIPS. In one embodiment, the selected at least three genes are MCM3, IRF7, and ULK1. In one embodiment, the selected at least three genes are MCM3, IRF7, and SPAG5. In one embodiment, the selected at least three genes are MCM3, BRIP1, and ULK1. In one embodiment, the selected at least three genes are MCM3, BRIP1, and SPAG5. In one embodiment, the selected at least three genes are MCM3, ULK1, and SPAG5. In one embodiment, the selected at least three genes are GPC1, MTUS1, and IRF7. In one embodiment, the selected at least three genes are GPC1, MTUS1, and BRIP1. In one embodiment, the selected at least three genes are GPC1, MTUS1, and ULK1. In one embodiment, the selected at least three genes are GPC1, MTUS1, and SPAG5. In one embodiment, the selected at least three genes are GPC1, IRF7, and BRIP1. In one embodiment, the selected at least three genes are GPC1, IRF7, and ULK1. In one embodiment, the selected at least three genes are GPC1, IRF7, and SPAG5. In one embodiment, the selected at least three genes are GPC1, BRIP1, and ULK1.In one embodiment, the selected at least three genes are GPC1, BRIP1, and SPAG5. In one embodiment, the selected at least three genes are GPC1, ULK1, and SPAG5. In one embodiment, the selected at least three genes are PCNA, IRF7, and BRIP1. In one embodiment, the selected at least three genes are PCNA, IRF7, and ULK1. In one embodiment, the selected at least three genes are PCNA, IRF7, and SPAG5. In one embodiment, the selected at least three genes are PCNA, BRIP1, and ULK1. In one embodiment, the selected at least three genes are PCNA, BRIP1, and SPAG5. In one embodiment, the selected at least three genes are PCNA, ULK1, and SPAG5. In one embodiment, the selected at least three genes are MTUS1, BRIP1, and UL1. In one embodiment, the selected at least three genes are MTUS1, BRIP1, and SPAG5. In one embodiment, the selected at least three genes are MTUS1, ULK1, and SPAG5. In one embodiment, the selected at least three genes are IRF7, ULK1, and SPAG5. In one embodiment, at least three genes are selected, and the selected genes are FOXM1, TTC21B, and CUL4A. In one embodiment, at least three genes are selected, and the selected genes are FOXM1, TTC21B, and MCM3. In one embodiment, at least three genes are selected, and the selected genes are FOXM1, TTC21B, and GPC1. In one embodiment, at least three genes are selected, and the selected genes are FOXM1, TTC21B, and PCNA. In one embodiment, at least three genes are selected, and the selected genes are FOXM1, TTC21B, and MTUS1. In one embodiment, at least three genes are selected, and the selected genes are FOXM1, TTC21B, and IRF7. In one embodiment, at least three genes are selected, and the selected genes are FOXM1, TTC21B, and BRIP1. In one embodiment, at least three genes are selected, and the selected genes are FOXM1, TTC21B, and ULK1. In one embodiment, at least three genes are selected, and the selected genes are FOXM1, TTC21B, and SPAG5. In one embodiment, at least three genes are selected, and the selected genes are CUL4, MCM3, and GPC1. In one embodiment, the selected at least three genes are FOXM1, MCM3, and CUL4. In one embodiment, the selected at least three genes are FOXM1, MCM3, and PCNA. In one embodiment, the selected at least three genes are FOXM1, MCM3, and MTUS1. In one embodiment, the selected at least three genes are FOXM1, MCM3, and IRF7. In one embodiment, the selected at least three genes are FOXM1, MCM3, and BRIP1.In one embodiment, the selected at least three genes are FOXM1, MCM3, and ULK1. In one embodiment, the selected at least three genes are FOXM1, MCM3, and SPAG5. In one embodiment, the selected at least three genes are FOXM1, GPC1, and PCNA. In one embodiment, the selected at least three genes are FOXM1, GPC1, and MTUS1. In one embodiment, the selected at least three genes are FOXM1, GPC1, and IRF7. In one embodiment, the selected at least three genes are FOXM1, GPC1, and BRIP1. In one embodiment, the selected at least three genes are FOXM1, GPC1, and ULK1. In one embodiment, the selected at least three genes are FOXM1, GPC1, and SPAG5. In one embodiment, the selected at least three genes are FOXM1, PCNA, and MTUS1. In one embodiment, the selected at least three genes are FOXM1, PCNA, and IRF7. In one embodiment, the selected at least three genes are FOXM1, PCNA, and BRIP1. In one embodiment, the selected at least three genes are FOXM1, PCNA, and ULK1. In one embodiment, the selected at least three genes are FOXM1, PCNA, and SPAG5. In one embodiment, the selected at least three genes are FOXM1, MTUS1, and IRF7. In one embodiment, the selected at least three genes are FOXM1, MTUS1, and BRIP1. In one embodiment, the selected at least three genes are FOXM1, MTUS1, and ULK1. In one embodiment, the selected at least three genes are FOXM1, MTUS1, and SPAG5. In one embodiment, the selected at least three genes are FOXM1, IRF7, and BRIP1. In one embodiment, the selected at least three genes are FOXM1, IRF7, and ULK1. In one embodiment, the selected at least three genes are FOXM1, IRF7, and SPAG5. In one embodiment, the selected at least three genes are FOXM1, BRIP1, and ULK1. In one embodiment, the selected at least three genes are FOXM1, BRIP1, and SPAG5. In one embodiment, the selected at least three genes are FOXM1, ULK1, and SPAG5.

[0031] In one embodiment, at least four genes are selected, and the selected at least four genes are CUL4A, TTC21B, MCM3, and GPC1. In one embodiment, the selected at least four genes are CUL4A, TTC21B, MCM3, and PCNA. In one embodiment, the selected at least four genes are CUL4A, TTC21B, MCM3, and MTUS1. In one embodiment, the selected at least four genes are CUL4A, TTC21B, MCM3, and IRF7. In one embodiment, the selected at least four genes are CUL4A, TTC21B, MCM3, and BRIP1. In one embodiment, the selected at least four genes are CUL4A, TTC21B, MCM3, and ULK1. In one embodiment, the selected at least four genes are CUL4A, TTC21B, MCM3, and SPAG5. In one embodiment, the selected at least four genes are CUL4A, TTC21B, GPC1, and PCNA. In one embodiment, the selected at least four genes are CUL4A, TTC21B, GPC1, and MTUS1. In one embodiment, the selected at least four genes are CUL4A, TTC21B, GPC1, and IRF7. In one embodiment, the selected at least four genes are CUL4A, TTC21B, GPC1, and BRIP1. In one embodiment, the selected at least four genes are CUL4A, TTC21B, GPC1, and ULK1. In one embodiment, the selected at least four genes are CUL4A, TTC21B, GPC1, and SPAG5. In one embodiment, the selected at least four genes are CUL4A, TTC21B, PCNA, and MTUS1. In one embodiment, the selected at least four genes are CUL4A, TTC21B, PCNA, and IRF7. In one embodiment, the selected at least four genes are CUL4A, TTC21B, PCNA, and BRIP1. In one embodiment, the selected at least four genes are CUL4A, TTC21B, PCNA, and ULK1. In one embodiment, the selected at least four genes are CUL4A, TTC21B, PCNA, and SPAG5. In one embodiment, the selected at least four genes are CUL4A, TTC21B, MTUS1, and IRF7. In one embodiment, the selected at least four genes are CUL4A, TTC21B, MTUS1, and BRIP1. In one embodiment, the selected at least four genes are CUL4A, TTC21B, MTUS1, and ULK1. In one embodiment, the selected at least four genes are CUL4A, TTC21B, MTUS1, and SPAG5. In one embodiment, the selected at least four genes are CUL4A, TTC21B, IRF7, and BRIP1.In one embodiment, the selected at least four genes are CUL4A, TTC21B, IRF7, and ULK1. In one embodiment, the selected at least four genes are CUL4A, TTC21B, IRF7, and SPAG5. In one embodiment, the selected at least four genes are CUL4A, TTC21B, BRIP1, and ULK1. In one embodiment, the selected at least four genes are CUL4A, TTC21B, BRIP1, and SPAG5. In one embodiment, the selected at least four genes are CUL4A, TTC21B, ULK1, and SPAG5. In one embodiment, the selected at least four genes are CUL4A, MCM3, GPC1, and PCNA. In one embodiment, the selected at least four genes are CUL4A, MCM3, GPC1, and MTUS1. In one embodiment, the selected at least four genes are CUL4A, MCM3, GPC1, and IRF7. In one embodiment, the selected at least four genes are CUL4A, MCM3, GPC1, and BRIP1. In one embodiment, the selected at least four genes are CUL4A, MCM3, GPC1, and ULK1. In one embodiment, the selected at least four genes are CUL4A, MCM3, GPC1, and SPAG5. In one embodiment, the selected at least four genes are CUL4A, MCM3, PCNA, and MTUS1. In one embodiment, the selected at least four genes are CUL4A, MCM3, PCNA, and IRF7. In one embodiment, the selected at least four genes are CUL4A, MCM3, PCNA, and BRIP1. In one embodiment, the selected at least four genes are CUL4A, MCM3, PCNA, and ULK1. In one embodiment, the selected at least four genes are CUL4A, MCM3, PCNA, and SPAG5. In one embodiment, the selected at least four genes are CUL4A, MCM3, MTUS1, and IRF7. In one embodiment, the selected at least four genes are CUL4A, MCM3, MTUS1, and BRIP1. In one embodiment, the selected at least four genes are CUL4A, MCM3, MTUS1, and ULK1. In one embodiment, the selected at least four genes are CUL4A, MCM3, MTUS1, and SPAG5. In one embodiment, the selected at least four genes are CUL4A, MCM3, IRF7, and BRIP1. In one embodiment, the selected at least four genes are CUL4A, MCM3, IRF7, and ULK1. In one embodiment, the selected at least four genes are CUL4A, MCM3, IRF7, and SPAG5. In one embodiment, the selected at least four genes are CUL4A, MCM3, BRIP1, and ULK1.In one embodiment, the selected at least four genes are CUL4A, MCM3, BRIP1, and SPAG5. In one embodiment, the selected at least four genes are CUL4A, MCM3, ULK1, and SPAG5. In one embodiment, the selected at least four genes are CUL4A, IRF7, BRIP1, and ULK1. In one embodiment, the selected at least four genes are CUL4A, IRF7, BRIP1, and SPAG5. In one embodiment, the selected at least four genes are CUL4A, BRIP1, ULK1, and SPAG5. In one embodiment, the selected at least four genes are TTC21B, MCM3, GPC1, and PCNA. In one embodiment, the selected at least four genes are TTC21B, MCM3, GPC1, and MTUS1. In one embodiment, the selected at least four genes are TTC21B, MCM3, GPC1, and IRF7. In one embodiment, the selected at least four genes are TTC21B, MCM3, GPC1, and BRIP1. In one embodiment, the selected at least four genes are TTC21B, MCM3, GPC1, and ULK1. In one embodiment, the selected at least four genes are TTC21B, MCM3, GPC1, and SPAG5. In one embodiment, the selected at least four genes are TTC21B, MCM3, PCNA, and MTUS1. In one embodiment, the selected at least four genes are TTC21B, MCM3, PCNA, and IRF7. In one embodiment, the selected at least four genes are TTC21B, MCM3, PCNA, and BRIP1. In one embodiment, the selected at least four genes are TTC21B, MCM3, PCNA, and ULK1. In one embodiment, the selected at least four genes are TTC21B, MCM3, PCNA, and SPAG5. In one embodiment, the selected at least four genes are TTC21B, MCM3, MTUS1, and IRF7. In one embodiment, the selected at least four genes are TTC21B, MCM3, MTUS1, and BRIP1. In one embodiment, the selected at least four genes are TTC21B, MCM3, MTUS1, and ULK1. In another embodiment, the selected at least four genes are TTC21B, MCM3, MTUS1, and SPAG5. In yet another embodiment, the selected at least four genes are TTC21B, MCM3, IRF7, and BRIP1. In yet another embodiment, the selected at least four genes are TTC21B, MCM3, IRF7, and ULK1. In yet another embodiment, the selected at least four genes are TTC21B, MCM3, IRF7, and SPAG5. In yet another embodiment, the selected at least four genes are TTC21B, MCM3, BRIP1, and ULK1.In one embodiment, the selected at least four genes are TTC21B, MCM3, IRF7, and SPAG5. In one embodiment, the selected at least four genes are TTC21B, MCM3, BRIP1, and ULK1. In one embodiment, the selected at least four genes are TTC21B, MCM3, IRF7, and SPAG5. In one embodiment, the selected at least four genes are TTC21B, MCM3, ULK1, and SPAG5. In one embodiment, the selected at least four genes are TTC21B, GPC1, PCNA, and MTUS1. In one embodiment, the selected at least four genes are TTC21B, GPC1, PCNA, and IRF7. In one embodiment, the selected at least four genes are TTC21B, GPC1, PCNA, and BRIP1. In one embodiment, the selected at least four genes are TTC21B, GPC1, PCNA, and ULK1. In one embodiment, the selected at least four genes are TTC21B, GPC1, PCNA, and SPAG5. In one embodiment, the selected at least four genes are TTC21B, PCNA, MTUS1, and IRF7. In another embodiment, the selected at least four genes are TTC21B, PCNA, MTUS1, and BRIP1. In yet another embodiment, the selected at least four genes are TTC21B, PCNA, MTUS1, and ULK1. In yet another embodiment, the selected at least four genes are TTC21B, PCNA, MTUS1, and SPAG5. In yet another embodiment, the selected at least four genes are TTC21B, MTUS1, IRF7, and BRIP1. In yet another embodiment, the selected at least four genes are TTC21B, MTUS1, IRF7, and ULK1. In yet another embodiment, the selected at least four genes are TTC21B, IRF7, BRIP1, and ULK1. In yet another embodiment, the selected at least four genes are TTC21B, IRF7, BRIP1, and SPAG5. In one embodiment, the selected at least four genes are MCM3, GPC1, PCNA, and MTUS1. In one embodiment, the selected at least four genes are MCM3, GPC1, PCNA, and IRF7. In one embodiment, the selected at least four genes are MCM3, GPC1, PCNA, and BRIP1. In one embodiment, the selected at least four genes are MCM3, GPC1, PCNA, and ULK1. In one embodiment, the selected at least four genes are MCM3, GPC1, PCNA, and SPAG5. In one embodiment, the selected at least four genes are MCM3, PCNA, MTUS1, and IRF7.In one embodiment, the selected at least four genes are MCM3, PCNA, MTUS1, and BRIP1. In one embodiment, the selected at least four genes are MCM3, PCNA, MTUS1, and ULK1. In one embodiment, the selected at least four genes are MCM3, PCNA, MTUS1, and SPAG5. In one embodiment, the selected at least four genes are MCM3, MTUS1, IRF7, and BRIP1. In one embodiment, the selected at least four genes are MCM3, MTUS1, IRF7, and ULK1. In one embodiment, the selected at least four genes are MCM3, MTUS1, IRF7, and SPAG5. In one embodiment, the selected at least four genes are MCM3, IRF7, BRIP1, and ULK1. In one embodiment, the selected at least four genes are MCM3, IRF7, BRIP1, and SPAG5. In one embodiment, the selected at least four genes are MCM3, BRIP1, ULK1, and SPAG5. In one embodiment, the selected at least four genes are GPC1, PCNA, MTUS1, and IRF7. In one embodiment, the selected at least four genes are GPC1, PCNA, MTUS1, and BRIP1. In another embodiment, the selected at least four genes are GPC1, PCNA, MTUS1, and ULK1. In yet another embodiment, the selected at least four genes are GPC1, PCNA, MTUS1, and SPAG5. In yet another embodiment, the selected at least four genes are GPC1, MTUS1, IRF7, and BRIP1. In yet another embodiment, the selected at least four genes are GPC1, MTUS1, IRF7, and ULK1. In yet another embodiment, the selected at least four genes are GPC1, IRF7, BRIP1, and ULK1. In yet another embodiment, the selected at least four genes are GPC1, IRF7, BRIP1, and SPAG5. In yet another embodiment, the selected at least four genes are GPC1, BRIP1, ULK1, and SPAG5. In yet another embodiment, the selected at least four genes are PCNA, MTUS1, BRIP1, and ULK1. In one embodiment, the selected at least four genes are PCNA, MTUS1, BRIP1, and SPAG5. In one embodiment, the selected at least four genes are PCNA, BRIP1, ULK1, and SPAG5. In one embodiment, the selected at least four genes are MTUS1, BRIP1, ULK1, and SPAG5. In one embodiment, the selected at least four genes are MTUS1, BRIP1, ULK1, and SPAG5.In one embodiment, at least four genes are selected, and the selected at least four genes are FOXM1, TTC21B, MCM3, and CUL4A. In one embodiment, at least four genes are selected, and the selected at least four genes are FOXM1, TTC21B, MCM3, and GPC1. In one embodiment, at least four genes are selected, and the selected at least four genes are FOXM1, TTC21B, MCM3, and PCNA. In one embodiment, at least four genes are selected, and the selected at least four genes are FOXM1, TTC21B, MCM3, and MTUS1. In one embodiment, at least four genes are selected, and the selected at least four genes are FOXM1, TTC21B, MCM3, and IRF7. In one embodiment, at least four genes are selected, and the selected at least four genes are FOXM1, TTC21B, MCM3, and BRIP1. In one embodiment, at least four genes are selected, and the selected at least four genes are FOXM1, TTC21B, MCM3, and SPAG5. In one embodiment, the selected at least four genes are FOXM1, TTC21B, GPC1, and PCNA. In another embodiment, the selected at least four genes are FOXM1, TTC21B, GPC1, and MTUS1. In yet another embodiment, the selected at least four genes are FOXM1, TTC21B, GPC1, and IRF7. In yet another embodiment, the selected at least four genes are FOXM1, TTC21B, GPC1, and BRIP1. In yet another embodiment, the selected at least four genes are FOXM1, TTC21B, GPC1, and ULK1. In yet another embodiment, the selected at least four genes are FOXM1, TTC21B, GPC1, and SPAG5. In yet another embodiment, the selected at least four genes are FOXM1, TTC21B, PCNA, and MTUS1. In yet another embodiment, the selected at least four genes are FOXM1, TTC21B, PCNA, and IRF7. In yet another embodiment, the selected at least four genes are FOXM1, TTC21B, PCNA, and BRIP1. In one embodiment, the selected at least four genes are FOXM1, TTC21B, PCNA, and ULK1. In one embodiment, the selected at least four genes are FOXM1, TTC21B, PCNA, and SPAG5. In one embodiment, the selected at least four genes are FOXM1, TTC21B, MTUS1, and IRF7. In one embodiment, the selected at least four genes are FOXM1, TTC21B, MTUS1, and BRIP1. In one embodiment, the selected at least four genes are FOXM1, TTC21B, MTUS1, and ULK1.In one embodiment, the selected at least four genes are FOXM1, TTC21B, MTUS1, and SPAG5. In one embodiment, the selected at least four genes are FOXM1, TTC21B, IRF7, and BRIP1. In one embodiment, the selected at least four genes are FOXM1, TTC21B, IRF7, and ULK1. In one embodiment, the selected at least four genes are FOXM1, TTC21B, IRF7, and SPAG5. In one embodiment, the selected at least four genes are FOXM1, TTC21B, BRIP1, and ULK1. In one embodiment, the selected at least four genes are FOXM1, TTC21B, BRIP1, and SPAG5. In one embodiment, the selected at least four genes are FOXM1, TTC21B, ULK1, and SPAG5. In one embodiment, the selected at least four genes are FOXM1, MCM3, GPC1, and PCNA. In one embodiment, the selected at least four genes are FOXM1, MCM3, GPC1, and MTUS1. In one embodiment, the selected at least four genes are FOXM1, MCM3, GPC1, and IRF7. In one embodiment, the selected at least four genes are FOXM1, MCM3, GPC1, and BRIP1. In one embodiment, the selected at least four genes are FOXM1, MCM3, GPC1, and ULK1. In one embodiment, the selected at least four genes are FOXM1, MCM3, GPC1, and SPAG5. In one embodiment, the selected at least four genes are FOXM1, MCM3, PCNA, and MTUS1. In one embodiment, the selected at least four genes are FOXM1, MCM3, PCNA, and IRF7. In one embodiment, the selected at least four genes are FOXM1, MCM3, PCNA, and BRIP1. In one embodiment, the selected at least four genes are FOXM1, MCM3, PCNA, and ULK1. In one embodiment, the selected at least four genes are FOXM1, MCM3, PCNA, and SPAG5. In one embodiment, the selected at least four genes are FOXM1, MCM3, MTUS1, and IRF7. In one embodiment, the selected at least four genes are FOXM1, MCM3, MTUS1, and BRIP1. In another embodiment, the selected at least four genes are FOXM1, MCM3, MTUS1, and ULK1. In yet another embodiment, the selected at least four genes are FOXM1, MCM3, MTUS1, and SPAG5. In yet another embodiment, the selected at least four genes are FOXM1, MCM3, IRF7, and BRIP1. In yet another embodiment, the selected at least four genes are FOXM1, MCM3, IRF7, and ULK1.In one embodiment, the selected at least four genes are FOXM1, MCM3, IRF7, and SPAG5. In another embodiment, the selected at least four genes are FOXM1, MCM3, BRIP1, and ULK1. In yet another embodiment, the selected at least four genes are FOXM1, MCM3, BRIP1, and SPAG5. In yet another embodiment, the selected at least four genes are FOXM1, MCM3, ULK1, and SPAG5. In yet another embodiment, the selected at least four genes are FOXM1, IRF7, BRIP1, and ULK1. In yet another embodiment, the selected at least four genes are FOXM1, IRF7, BRIP1, and SPAG5. In yet another embodiment, the selected at least four genes are FOXM1, BRIP1, ULK1, and SPAG5.

[0032] In one embodiment, at least five genes are selected, and the selected genes are CUL4, TTC21B, MCM3, GPC1, and PCNA. In another embodiment, the selected at least five genes are CUL4, TTC21B, MCM3, GPC1, and MTUS1. In yet another embodiment, the selected at least five genes are CUL4, TTC21B, MCM3, GPC1, and PCNA. In yet another embodiment, the selected at least five genes are CUL4, TTC21B, MCM3, GPC1, and MTUS1. In yet another embodiment, the selected at least five genes are CUL4, TTC21B, MCM3, GPC1, and IRF7. In yet another embodiment, the selected at least five genes are CUL4, TTC21B, MCM3, GPC1, and BRIP1. In yet another embodiment, the selected at least five genes are CUL4, TTC21B, MCM3, GPC1, and ULK1. In yet another embodiment, the selected at least five genes are CUL4, TTC21B, MCM3, GPC1, and SPAG5. In one embodiment, the selected at least five genes are CUL4, MCM3, GPC1, PCNA, and MTUS1. In one embodiment, the selected at least five genes are CUL4, MCM3, GPC1, PCNA, and IRF7. In one embodiment, the selected at least five genes are CUL4, MCM3, GPC1, PCNA, and BRIP1. In one embodiment, the selected at least five genes are CUL4, MCM3, GPC1, PCNA, and ULK1. In one embodiment, the selected at least five genes are CUL4, MCM3, GPC1, PCNA, and SPAG5. In one embodiment, the selected at least five genes are CUL4, GPC1, PCNA, MTUS1, and IRF7. In one embodiment, the selected at least five genes are CUL4, GPC1, PCNA, MTUS1, and BRIP1. In one embodiment, the selected at least five genes are CUL4, GPC1, PCNA, MTUS1, and ULK1. In one embodiment, the selected at least five genes are CUL4, GPC1, PCNA, MTUS1, and SPAG5. In one embodiment, the selected at least five genes are CUL4, PCNA, MTUS1, IRF7, and BRIP1. In another embodiment, the selected at least five genes are CUL4, PCNA, MTUS1, IRF7, and ULK1. In yet another embodiment, the selected at least five genes are CUL4, PCNA, MTUS1, IRF7, and SPAG5. In yet another embodiment, the selected at least five genes are CUL4, MTUS1, IRF7, and BRIP1.

[0033] and ULK1. In one embodiment, the selected at least five genes are CUL4, MTUS1, IRF7, BRIP1, and ULK1.

[0034] and SPAG5. In one embodiment, the selected at least five genes are CUL4, IRF7, BRIP1, ULK1, and

[0035] and SPAG5. In one embodiment, the selected at least five genes are TTC21B, MCM3, GPC1, PCNA, and

[0036] and MTUS1. In one embodiment, the selected at least five genes are TTC21B, MCM3, GPC1, PCNA, and MTUS1.

[0037] and IRF7. In one embodiment, the selected at least five genes are TTC21B, MCM3, GPC1, PCNA, and

[0038] and BRIP1. In one embodiment, the selected at least five genes are TTC21B, MCM3, GPC1, PCNA, and BRIP1.

[0039] and ULK1. In one embodiment, the selected at least five genes are TTC21B, MCM3, GPC1, PCNA

[0040] and SPAG5. In one embodiment, the selected at least five genes are TTC21B, GPC1, PCNA, MTUS1, and SPAG5.

[0041] and IRF7. In one embodiment, the selected at least five genes are TTC21B, GPC1, PCNA, MTUS1, and IRF7.

[0042] and BRIP1. In one embodiment, the selected at least five genes are TTC21B, GPC1, PCNA, MTUS1, and BRIP1.

[0043] and ULK1. In one embodiment, the selected at least five genes are TTC21B, GPC1, PCNA, MTUS1, and ULK1.

[0044] and SPAG5. In one embodiment, the selected at least five genes are TTC21B, PCNA, MTUS1, IRF7, and

[0045] and BRIP1. In one embodiment, the selected at least five genes are TTC21B, PCNA, MTUS1, IRF7, and BRIP1.

[0046] and ULK1. In one embodiment, the selected at least five genes are TTC21B, PCNA, MTUS1, IRF7, and ULK1.

[0047] and SPAG5. In one embodiment, the selected at least five genes are TTC21B, MTUS1, IRF7, BRIP1, and SPAG5.

[0048] and ULK1. In one embodiment, the selected at least five genes are TTC21B, MTUS1, IRF7, BRIP1, and ULK1.

[0049] and SPAG5. In one embodiment, the selected at least five genes are TTC21B, IRF7, BRIP1, ULK1, and

[0050] and SPAG5. In one embodiment, the selected at least five genes are MCM3, GPC1, PCNA, MTUS1, and SPAG5.

[0051] and IRF7. In one embodiment, the selected at least five genes are MCM3, GPC1, PCNA, MTUS1, and IRF7.

[0052] and IRF7. In one embodiment, the selected at least five genes are MCM3, GPC1, PCNA, MTUS1, and IRF7.

[0053] and ULK1. In one embodiment, the selected at least five genes are MCM3, GPC1, PCNA, MTUS1, and ULK1.

[0054] and SPAG5. In one embodiment, the selected at least five genes are MCM3, PCNA, MTUS1, IRF7, and

[0055] and BRIP1. In one embodiment, the selected at least five genes are MCM3, PCNA, MTUS1, IRF7, and BRIP1.

[0056] and ULK1. In one embodiment, the selected at least five genes are MCM3, PCNA, MTUS1, IRF7, and ULK1.

[0057] and SPAG5. In one embodiment, the selected at least five genes are MCM3, MTUS1, IRF7, BRIP1, and SPAG5.

[0058] and ULK1. In one embodiment, the selected at least five genes are MCM3, MTUS1, IRF7, BRIP1, and ULK1.

[0059] and SPAG5. In one embodiment, the selected at least five genes are MCM3, IRF7, BRIP1, ULK1, and

[0060] and SPAG5. In one embodiment, the selected at least five genes are GPC1, PCNA, MTUS1, IRF7, and

[0061] and BRIP1. In one embodiment, the selected at least five genes are GPC1, PCNA, MTUS1, IRF7, and BRIP1.

[0062] And ULK1. In one embodiment, the selected at least five genes are GPC1, PCNA, MTUS1, IRF7, and SPAG5. In one embodiment, the selected at least five genes are GPC1, MTUS1, IRF7, BRIP1, and ULK1. In one embodiment, the selected at least five genes are GPC1, MTUS1, IRF7, BRIP1, and SPAG5. In one embodiment, the selected at least five genes are GPC1, IRF7, BRIP1, ULK1, and SPAG5. In one embodiment, the selected at least five genes are PCNA, MTUS1, IRF7, BRIP1, ULK1, and SPAG5. In one embodiment, the selected at least five genes are PCNA, IRF7, BRIP1, ULK1, and SPAG5. In one embodiment, the selected at least five genes are MTUS1, IRF7, BRIP1, ULK1, and SPAG5. In one embodiment, at least five genes are selected, and the selected genes are FOXM1, TTC21B, MCM3, GPC1, and CUL4. In another embodiment, at least five genes are selected, and the selected genes are FOXM1, TTC21B, MCM3, GPC1, and PCNA. In yet another embodiment, at least five genes are selected, and ... In one embodiment, the selected at least five genes are FOXM1, TTC21B, MCM3, GPC1, and ULK1. In one embodiment, the selected at least five genes are FOXM1, TTC21B, MCM3, GPC1, and SPAG5. In one embodiment, the selected at least five genes are FOXM1, MCM3, GPC1, PCNA, and MTUS1. In one embodiment, the selected at least five genes are FOXM1, MCM3, GPC1, PCNA, and IRF7. In one embodiment, the selected at least five genes are FOXM1, MCM3, GPC1, PCNA, and BRIP1. In one embodiment, the selected at least five genes are FOXM1, MCM3, GPC1, PCNA, and ULK1.In one embodiment, the selected at least five genes are FOXM1, MCM3, GPC1, PCNA, and SPAG5. In one embodiment, the selected at least five genes are FOXM1, GPC1, PCNA, MTUS1, and IRF7. In one embodiment, the selected at least five genes are FOXM1, GPC1, PCNA, MTUS1, and BRIP1. In one embodiment, the selected at least five genes are FOXM1, GPC1, PCNA, MTUS1, and ULK1. In one embodiment, the selected at least five genes are FOXM1, GPC1, PCNA, MTUS1, and SPAG5. In one embodiment, the selected at least five genes are FOXM1, PCNA, MTUS1, IRF7, and BRIP1. In one embodiment, the selected at least five genes are FOXM1, PCNA, MTUS1, IRF7, and ULK1.

[0063] In one embodiment, the selected at least five genes are FOXM1, PCNA, MTUS1, IRF7, and SPAG5.

[0064] In one embodiment, the selected at least five genes are FOXM1, MTUS1, IRF7, BRIP1, and ULK1.

[0065] In one embodiment, the selected at least five genes are FOXM1, MTUS1, IRF7, BRIP1, and SPAG5.

[0066] In one embodiment, the selected at least five genes are FOXM1, IRF7, BRIP1, ULK1, and SPAG5.

[0067] In one embodiment, at least six genes are selected, and the selected genes are CUL4A, TTC21B, MCM3, GPC1, PCNA, and MTUS1. In one embodiment, the selected at least six genes are CUL4A, TTC21B, MCM3, GPC1, PCNA, and IRF7. In one embodiment, the selected at least six genes are CUL4A, TTC21B, MCM3, GPC1, PCNA, and BRIP1. In one embodiment, the selected at least six genes are CUL4A, TTC21B, MCM3, GPC1, PCNA, and ULK1. In one embodiment, the selected at least six genes are CUL4A, TTC21B, MCM3, GPC1, PCNA, and SPAG5. In one embodiment, the selected at least six genes are CUL4A, MCM3, GPC1, PCNA, MTUS1, and IRF7. In one embodiment, the selected at least six genes are CUL4A, MCM3, GPC1, PCNA, MTUS1, and BRIP1. In one embodiment, the selected at least six genes are CUL4A, MCM3, GPC1, PCNA, MTUS1, and ULK1. In another embodiment, the selected at least six genes are CUL4A, MCM3, GPC1, PCNA, MTUS1, and SPAG5. In yet another embodiment, the selected at least six genes are CUL4A, GPC1, PCNA, MTUS1, IRF7, and BRIP1. In yet another embodiment, the selected at least six genes are CUL4A, GPC1, PCNA, MTUS1, IRF7, and ULK1. In yet another embodiment, the selected at least six genes are CUL4A, GPC1, PCNA, MTUS1, IRF7, and SPAG5. In yet another embodiment, the selected at least six genes are CUL4A, PCNA, MTUS1, IRF7, BRIP1, and ULK1. In yet another embodiment, the selected at least six genes are CUL4A, PCNA, MTUS1, IRF7, BRIP1, and SPAG5. In one embodiment, the selected at least six genes are CUL4A, MTUS1, IRF7, BRIP1, ULK1, and SPAG5. In one embodiment, the selected at least six genes are TTC21B, MCM3, GPC1, PCNA, MTUS1, and IRF7. In one embodiment, the selected at least six genes are TTC21B, MCM3, GPC1, PCNA, MTUS1, and BRIP1. In one embodiment, the selected at least six genes are TTC21B, MCM3, GPC1, PCNA, MTUS1, and ULK1. In one embodiment, the selected at least six genes are TTC21B, MCM3, GPC1, PCNA, MTUS1, and SPAG5.In one embodiment, the selected at least six genes are TTC21B, GPC1, PCNA, MTUS1, IRF7, and BRIP1. In another embodiment, the selected at least six genes are TTC21B, GPC1, PCNA, MTUS1, IRF7, and ULK1. In yet another embodiment, the selected at least six genes are TTC21B, GPC1, PCNA, MTUS1, IRF7, and SPAG5. In yet another embodiment, the selected at least six genes are TTC21B, PCNA, MTUS1, IRF7, BRIP1, and ULK1. In yet another embodiment, the selected at least six genes are TTC21B, PCNA, MTUS1, IRF7, BRIP1, ULK1, and SPAG5. In yet another embodiment, the selected at least six genes are TTC21B, MTUS1, IRF7, BRIP1, ULK1, and SPAG5. In yet another embodiment, the selected at least six genes are TTC21B, MTUS1, IRF7, BRIP1, ULK1, and SPAG5. In one embodiment, the selected at least six genes are TTC21B, MCM3, GPC1, PCNA, IRF7, and BRIP1. In another embodiment, the selected at least six genes are TTC21B, MCM3, GPC1, PCNA, IRF7, and ULK1. In yet another embodiment, the selected at least six genes are TTC21B, MCM3, GPC1, PCNA, IRF7, and SPAG5. In yet another embodiment, the selected at least six genes are TTC21B, GPC1, PCNA, IRF7, BRIP1, and ULK1. In yet another embodiment, the selected at least six genes are TTC21B, GPC1, PCNA, IRF7, BRIP1, ULK1, and SPAG5. In yet another embodiment, the selected at least six genes are TTC21B, PCNA, IRF7, BRIP1, ULK1, and SPAG5. In yet another embodiment, the selected at least six genes are MCM3, GPC1, PCNA, MTUS1, IRF7, and BRIP1. In one embodiment, the selected at least six genes are MCM3, GPC1, PCNA, MTUS1, IRF7, and ULK1. In another embodiment, the selected at least six genes are MCM3, GPC1, PCNA, MTUS1, IRF7, and SPAG5. In yet another embodiment, the selected at least six genes are MCM3, PCNA, MTUS1, IRF7, BRIP1, and ULK1. In yet another embodiment, the selected at least six genes are MCM3, GPC1, PCNA, MTUS1, IRF7, and SPAG5. In yet another embodiment, the selected at least six genes are MCM3, PCNA, MTUS1, IRF7, BRIP1, and ULK1.In one embodiment, the selected at least six genes are MCM3, PCNA, MTUS1, IRF7, BRIP1, and SPAG5. In one embodiment, the selected at least six genes are GPC1, PCNA, MTUS1, IRF7, BRIP1, and ULK1. In one embodiment, the selected at least six genes are GPC1, PCNA, MTUS1, IRF7, BRIP1, and SPAG5. In one embodiment, the selected at least six genes are PCNA, MTUS1, IRF7, BRIP1, ULK1, and SPAG5. In one embodiment, at least six genes are selected, and the selected genes are FOXM1, TTC21B, MCM3, GPC1, PCNA, and CUL4A. In one embodiment, at least six genes are selected, and the selected genes are FOXM1, TTC21B, MCM3, GPC1, PCNA, and MTUS1. In one embodiment, at least six genes are selected, and the selected genes are FOXM1, TTC21B, MCM3, GPC1, PCNA, and IRF7. In one embodiment, the selected at least six genes are FOXM1, TTC21B, MCM3, GPC1, PCNA, and BRIP1. In another embodiment, the selected at least six genes are FOXM1, TTC21B, MCM3, GPC1, PCNA, and ULK1. In yet another embodiment, the selected at least six genes are FOXM1, TTC21B, MCM3, GPC1, PCNA, and SPAG5. In yet another embodiment, the selected at least six genes are FOXM1, MCM3, GPC1, PCNA, MTUS1, and IRF7. In yet another embodiment, the selected at least six genes are FOXM1, MCM3, GPC1, PCNA, MTUS1, and BRIP1. In yet another embodiment, the selected at least six genes are FOXM1, MCM3, GPC1, PCNA, MTUS1, and ULK1. In yet another embodiment, the selected at least six genes are FOXM1, MCM3, GPC1, PCNA, MTUS1, and SPAG5. In one embodiment, the selected at least six genes are FOXM1, GPC1, PCNA, MTUS1, IRF7, and BRIP1. In another embodiment, the selected at least six genes are FOXM1, GPC1, PCNA, MTUS1, IRF7, and ULK1. In yet another embodiment, the selected at least six genes are FOXM1, GPC1, PCNA, MTUS1, IRF7, and SPAG5. In yet another embodiment, the selected at least six genes are FOXM1, PCNA, MTUS1, IRF7, BRIP1, and ULK1.In one embodiment, the selected at least six genes are FOXM1, PCNA, MTUS1, IRF7, BRIP1, and SPAG5. In another embodiment, the selected at least six genes are FOXM1, MTUS1, IRF7, BRIP1, ULK1, and SPAG5.

[0068] In one embodiment, at least seven genes are selected, and the selected genes are CUL4A, TTC21B, MCM3, GPC1, PCNA, MTUS1, and IRF7. In another embodiment, the selected at least seven genes are CUL4A, TTC21B, MCM3, GPC1, PCNA, MTUS1, and IRF7. In yet another embodiment, the selected at least seven genes are CUL4A, TTC21B, MCM3, GPC1, PCNA, MTUS1, and BRIP1. In yet another embodiment, the selected at least seven genes are CUL4A, TTC21B, MCM3, GPC1, PCNA, MTUS1, and ULK1. In yet another embodiment, the selected at least seven genes are CUL4A, TTC21B, MCM3, GPC1, PCNA, MTUS1, and SPAG5. In yet another embodiment, the selected at least seven genes are CUL4A, MCM3, GPC1, PCNA, MTUS1, IRF7, and BRIP1. In one embodiment, the selected at least seven genes are CUL4A, MCM3, GPC1, PCNA, MTUS1, IRF7, and ULK1. In another embodiment, the selected at least seven genes are CUL4A, MCM3, GPC1, PCNA, MTUS1, IRF7, and SPAG5. In yet another embodiment, the selected at least seven genes are CUL4A, GPC1, PCNA, MTUS1, IRF7, BRIP1, and ULK1. In yet another embodiment, the selected at least seven genes are CUL4A, GPC1, PCNA, MTUS1, IRF7, BRIP1, and SPAG5. In yet another embodiment, the selected at least seven genes are CUL4A, PCNA, MTUS1, IRF7, BRIP1, ULK1, and SPAG5. In yet another embodiment, the selected at least seven genes are TTC21B, MCM3, GPC1, PCNA, MTUS1, IRF7, and BRIP1. In one embodiment, the selected at least seven genes are TTC21B, MCM3, GPC1, PCNA, MTUS1, IRF7, and ULK1. In another embodiment, the selected at least seven genes are TTC21B, MCM3, GPC1, PCNA, MTUS1, IRF7, and SPAG5. In yet another embodiment, the selected at least seven genes are TTC21B, GPC1, PCNA, MTUS1, IRF7, BRIP1, and ULK1. In yet another embodiment, the selected at least seven genes are TTC21B, GPC1, PCNA, MTUS1, IRF7, BRIP1, ULK1, and SPAG5. In yet another embodiment, the selected at least seven genes are TTC21B, PCNA, MTUS1, IRF7, BRIP1, ULK1, and SPAG5.In one embodiment, the selected at least seven genes are MCM3, GPC1, PCNA, MTUS1, IRF7, BRIP1, and ULK1. In another embodiment, the selected at least seven genes are MCM3, GPC1, PCNA, MTUS1, IRF7, BRIP1, and SPAG5. In yet another embodiment, the selected at least seven genes are MCM3, PCNA, MTUS1, IRF7, BRIP1, ULK1, and SPAG5. In yet another embodiment, the selected at least seven genes are GPC1, PCNA, MTUS1, IRF7, BRIP1, ULK1, and SPAG5. In yet another embodiment, at least seven genes are selected, and the selected genes are FOXM1, TTC21B, MCM3, GPC1, PCNA, MTUS1, and CUL4A. In yet another embodiment, at least seven genes are selected, and the selected genes are FOXM1, TTC21B, MCM3, GPC1, PCNA, MTUS1, and IRF7. In one embodiment, the selected at least seven genes are FOXM1, TTC21B, MCM3, GPC1, PCNA, MTUS1, and IRF7. In another embodiment, the selected at least seven genes are FOXM1, TTC21B, MCM3, GPC1, PCNA, MTUS1, and BRIP1. In yet another embodiment, the selected at least seven genes are FOXM1, TTC21B, MCM3, GPC1, PCNA, MTUS1, and ULK1. In yet another embodiment, the selected at least seven genes are FOXM1, TTC21B, MCM3, GPC1, PCNA, MTUS1, and SPAG5. In yet another embodiment, the selected at least seven genes are FOXM1A, MCM3, GPC1, PCNA, MTUS1, IRF7, and BRIP1. In yet another embodiment, the selected at least seven genes are FOXM1, MCM3, GPC1, PCNA, MTUS1, IRF7, and ULK1. In one embodiment, the selected at least seven genes are FOXM1, MCM3, GPC1, PCNA, MTUS1, IRF7, and SPAG5. In another embodiment, the selected at least seven genes are FOXM1, GPC1, PCNA, MTUS1, IRF7, BRIP1, and ULK1. In yet another embodiment, the selected at least seven genes are FOXM1, GPC1, PCNA, MTUS1, IRF7, BRIP1, ULK1, and SPAG5. In yet another embodiment, the selected at least seven genes are FOXM1, PCNA, MTUS1, IRF7, BRIP1, ULK1, and SPAG5.

[0069] In one embodiment, at least eight genes are selected, and the selected genes are CUL4A, TTC21B, MCM3, GPC1, PCNA, MTUS1, IRF7, and BRIP1. In another embodiment, the selected at least eight genes are CUL4A, TTC21B, MCM3, GPC1, PCNA, MTUS1, IRF7, and ULK1. In yet another embodiment, the selected at least eight genes are CUL4A, TTC21B, MCM3, GPC1, PCNA, MTUS1, IRF7, and SPAG5. In yet another embodiment, the selected at least eight genes are CUL4A, MCM3, GPC1, PCNA, MTUS1, IRF7, BRIP1, and ULK1. In yet another embodiment, the selected at least eight genes are CUL4A, MCM3, GPC1, PCNA, MTUS1, IRF7, BRIP1, and SPAG5. In one embodiment, the selected at least eight genes are CUL4A, GPC1, PCNA, MTUS1, IRF7, BRIP1, ULK1, and SPAG5. In another embodiment, the selected at least eight genes are CUL4A, TTC21B, GPC1, PCNA, MTUS1, IRF7, BRIP1, and ULK1. In yet another embodiment, the selected at least eight genes are CUL4A, TTC21B, GPC1, PCNA, MTUS1, IRF7, BRIP1, and SPAG5. In yet another embodiment, the selected at least eight genes are CUL4A, TTC21B, GPC1, PCNA, MTUS1, IRF7, ULK1, and SPAG5. In yet another embodiment, the selected at least eight genes are CUL4A, TTC21B, MCM3, PCNA, MTUS1, IRF7, ULK1, and SPAG5. In one embodiment, the selected at least eight genes are CUL4A, TTC21B, MCM3, GPC1, MTUS1, IRF7, ULK1, and SPAG5. In another embodiment, the selected at least eight genes are CUL4A, TTC21B, MCM3, GPC1, PCNA, IRF7, ULK1, and SPAG5. In yet another embodiment, the selected at least eight genes are CUL4A, TTC21B, MCM3, GPC1, PCNA, MTUS1, ULK1, and SPAG5. In yet another embodiment, the selected at least eight genes are CUL4A, TTC21B, MCM3, GPC1, PCNA, MTUS1, IRF7, and ULK1. In yet another embodiment, the selected at least eight genes are CUL4A, MCM3, GPC1, PCNA, MTUS1, IRF7, ULK1, and SPAG5.In one embodiment, the selected at least eight genes are CUL4A, GPC1, PCNA, MTUS1, IRF7, BRIP1, ULK1, and SPAG5. In another embodiment, the selected at least eight genes are CUL4A, TTC21B, PCNA, MTUS1, IRF7, BRIP1, ULK1, and SPAG5. In yet another embodiment, the selected at least eight genes are CUL4A, TTC21B, MCM3, MTUS1, IRF7, BRIP1, ULK1, and SPAG5. In yet another embodiment, the selected at least eight genes are CUL4A, TTC21B, MCM3, GPC1, IRF7, BRIP1, ULK1, and SPAG5. In one embodiment, the selected at least eight genes are CUL4A, TTC21B, MCM3, GPC1, PCNA, MTUS1, ULK1, and SPAG5. In another embodiment, the selected at least eight genes are CUL4A, TTC21B, MCM3, GPC1, PCNA, MTUS1, IRF7, and SPAG5. In yet another embodiment, the selected at least eight genes are TTC21B, MCM3, GPC1, PCNA, MTUS1, IRF7, BRIP1, and ULK1. In yet another embodiment, the selected at least eight genes are TTC21B, MCM3, GPC1, PCNA, MTUS1, IRF7, BRIP1, and SPAG5. In yet another embodiment, the selected at least eight genes are TTC21B, GPC1, PCNA, MTUS1, IRF7, BRIP1, ULK1, and SPAG5. In one embodiment, the selected at least eight genes are TTC21B, MCM3, PCNA, MTUS1, IRF7, BRIP1, ULK1, and SPAG5. In another embodiment, the selected at least eight genes are TTC21B, MCM3, GPC1, MTUS1, IRF7, BRIP1, ULK1, and SPAG5. In yet another embodiment, the selected at least eight genes are TTC21B, MCM3, GPC1, PCNA, IRF7, BRIP1, ULK1, and SPAG5. In yet another embodiment, the selected at least eight genes are TTC21B, MCM3, GPC1, PCNA, MTUS1, BRIP1, ULK1, and SPAG5.In one embodiment, the selected at least eight genes are TTC21B, MCM3, GPC1, PCNA, MTUS1, IRF7, BRIP1, and ULK1. In another embodiment, the selected at least eight genes are TTC21B, MCM3, GPC1, PCNA, MTUS1, IRF7, BRIP1, and SPAG5. In yet another embodiment, the selected at least eight genes are MCM3, GPC1, PCNA, MTUS1, IRF7, BRIP1, ULK1, and SPAG5. In yet another embodiment, at least eight genes are selected, and the selected genes are FOXM1, TTC21B, MCM3, GPC1, PCNA, MTUS1, IRF7, and CUL4A. In yet another embodiment, at least eight genes are selected, and the selected genes are FOXM1, TTC21B, MCM3, GPC1, PCNA, MTUS1, IRF7, and BRIP1. In one embodiment, the selected at least eight genes are FOXM1, TTC21B, MCM3, GPC1, PCNA, MTUS1, IRF7, and ULK1. In another embodiment, the selected at least eight genes are FOXM1, TTC21B, MCM3, GPC1, PCNA, MTUS1, IRF7, and SPAG5. In yet another embodiment, the selected at least eight genes are FOXM1, MCM3, GPC1, PCNA, MTUS1, IRF7, BRIP1, and ULK1. In yet another embodiment, the selected at least eight genes are FOXM1, MCM3, GPC1, PCNA, MTUS1, IRF7, BRIP1, and SPAG5. In yet another embodiment, the selected at least eight genes are FOXM1, GPC1, PCNA, MTUS1, IRF7, BRIP1, ULK1, and SPAG5. In yet another embodiment, the selected at least eight genes are FOXM1, TTC21B, GPC1, PCNA, MTUS1, IRF7, BRIP1, and ULK1. In one embodiment, the selected at least eight genes are FOXM1, TTC21B, GPC1, PCNA, MTUS1, IRF7, BRIP1, and SPAG5. In another embodiment, the selected at least eight genes are FOXM1, TTC21B, GPC1, PCNA, MTUS1, IRF7, ULK1, and SPAG5. In yet another embodiment, the selected at least eight genes are FOXM1, TTC21B, MCM3, PCNA, MTUS1, IRF7, ULK1, and SPAG5.In one embodiment, the selected at least eight genes are FOXM1, TTC21B, MCM3, GPC1, PCNA, IRF7, ULK1, and SPAG5. In another embodiment, the selected at least eight genes are FOXM1, TTC21B, MCM3, GPC1, PCNA, MTUS1, ULK1, and SPAG5. In yet another embodiment, the selected at least eight genes are FOXM1, TTC21B, MCM3, GPC1, PCNA, MTUS1, IRF7, and ULK1. In yet another embodiment, the selected at least eight genes are FOXM1, MCM3, GPC1, PCNA, MTUS1, IRF7, ULK1, and SPAG5. In yet another embodiment, the selected at least eight genes are FOXM1, GPC1, PCNA, MTUS1, IRF7, BRIP1, ULK1, and SPAG5. In one embodiment, the selected at least eight genes are FOXM1, TTC21B, PCNA, MTUS1, IRF7, BRIP1, ULK1, and SPAG5. In another embodiment, the selected at least eight genes are FOXM1, TTC21B, MCM3, MTUS1, IRF7, BRIP1, ULK1, and SPAG5. In yet another embodiment, the selected at least eight genes are FOXM1, TTC21B, MCM3, GPC1, IRF7, BRIP1, ULK1, and SPAG5. In yet another embodiment, the selected at least eight genes are FOXM1, TTC21B, MCM3, GPC1, PCNA, BRIP1, ULK1, and SPAG5. In yet another embodiment, the selected at least eight genes are FOXM1, TTC21B, MCM3, GPC1, PCNA, MTUS1, ULK1, and SPAG5. In one embodiment, the selected at least eight genes are FOXM1, TTC21B, MCM3, GPC1, PCNA, MTUS1, IRF7, and SPAG5.

[0070] In one embodiment, at least nine genes are selected, and the selected genes are CUL4A, TTC21B, MCM3, GPC1, PCNA, MTUS1, IRF7, BRIP1, and ULK1. In another embodiment, the selected at least nine genes are CUL4A, TTC21B, MCM3, GPC1, PCNA, MTUS1, IRF7, BRIP1, and SPAG5. In yet another embodiment, the selected at least nine genes are CUL4A, TTC21B, GPC1, PCNA, MTUS1, IRF7, BRIP1, ULK1, and SPAG5. In yet another embodiment, the selected at least nine genes are CUL4A, TTC21B, GPC1, PCNA, MTUS1, IRF7, BRIP1, ULK1, and SPAG5. In one embodiment, the selected at least nine genes are CUL4A, TTC21B, MCM3, GPC1, MTUS1, IRF7, BRIP1, ULK1, and SPAG5. In another embodiment, the selected at least nine genes are CUL4A, TTC21B, MCM3, GPC1, PCNA, IRF7, BRIP1, ULK1, and SPAG5. In another embodiment, the selected at least nine genes are CUL4A, TTC21B, MCM3, GPC1, PCNA, MTUS1, BRIP1, ULK1, and SPAG5. In another embodiment, the selected at least nine genes are CUL4A, TTC21B, MCM3, GPC1, PCNA, MTUS1, IRF7, ULK1, and SPAG5. In another embodiment, the selected at least nine genes are CUL4A, TTC21B, MCM3, GPC1, PCNA, MTUS1, IRF7, BRIP1, and SPAG5. In one embodiment, the selected at least nine genes are CUL4A, TTC21B, MCM3, GPC1, PCNA, MTUS1, IRF7, BRIP1, and ULK1. In another embodiment, at least nine genes are selected, and the selected genes are FOXM1, TTC21B, MCM3, GPC1, PCNA, MTUS1, IRF7, BRIP1, and CUL4A. In another embodiment, the selected at least nine genes are FOXM1, TTC21B, MCM3, GPC1, PCNA, MTUS1, IRF7, BRIP1, and SPAG5. In yet another embodiment, the selected at least nine genes are FOXM1, TTC21B, GPC1, PCNA, MTUS1, IRF7, BRIP1, ULK1, and SPAG5.In one embodiment, the selected at least nine genes are FOXM1, TTC21B, MCM3, PCNA, MTUS1, IRF7, BRIP1, ULK1, and SPAG5. In another embodiment, the selected at least nine genes are FOXM1, TTC21B, MCM3, GPC1, MTUS1, IRF7, BRIP1, ULK1, and SPAG5. In yet another embodiment, the selected at least nine genes are FOXM1, TTC21B, MCM3, GPC1, PCNA, IRF7, BRIP1, ULK1, and SPAG5. In yet another embodiment, the selected at least nine genes are FOXM1, TTC21B, MCM3, GPC1, PCNA, MTUS1, BRIP1, ULK1, and SPAG5. In one embodiment, the selected at least nine genes are FOXM1, TTC21B, MCM3, GPC1, PCNA, MTUS1, IRF7, BRIP1, and SPAG5. In another embodiment, the selected at least nine genes are FOXM1, TTC21B, MCM3, GPC1, PCNA, MTUS1, IRF7, BRIP1, and ULK1.

[0071] In one embodiment, at least ten genes are selected, and the selected genes are CUL4A, TTC21B, MCM3, GPC1, PCNA, MTUS1, IRF7, BRIP1, ULK1, and SPAG5. In another embodiment, the selected at least ten genes are FOXM1, CUL4A, TTC21B, MCM3, GPC1, PCNA, MTUS1, IRF7, BRIP1, and ULK1. In yet another embodiment, the selected at least ten genes are FOXM1, CUL4A, TTC21B, MCM3, GPC1, PCNA, MTUS1, IRF7, BRIP1, and SPAG5. In yet another embodiment, the selected at least ten genes are FOXM1, CUL4A, TTC21B, MCM3, GPC1, PCNA, MTUS1, IRF7, ULK1, and SPAG5. In one embodiment, the selected at least ten genes are FOXM1, CUL4A, TTC21B, MCM3, GPC1, PCNA, MTUS1, BRIP1, ULK1, and SPAG5. In another embodiment, the selected at least ten genes are FOXM1, CUL4A, TTC21B, MCM3, GPC1, PCNA, IRF7, BRIP1, ULK1, and SPAG5. In yet another embodiment, the selected at least ten genes are FOXM1, CUL4A, TTC21B, MCM3, GPC1, MTUS1, IRF7, BRIP1, ULK1, and SPAG5. In one embodiment, the selected at least ten genes are FOXM1, CUL4A, TTC21B, MCM3, PCNA, MTUS1, IRF7, BRIP1, ULK1, and SPAG5. In another embodiment, the selected at least ten genes are FOXM1, CUL4A, TTC21B, GPC1, PCNA, MTUS1, IRF7, BRIP1, ULK1, and SPAG5. In yet another embodiment, the selected at least ten genes are FOXM1, CUL4A, MCM3, GPC1, PCNA, MTUS1, IRF7, BRIP1, ULK1, and SPAG5.

[0072] In one embodiment, all of TTC21B, MCM3, MTUS1, FOXM1, CUL4A, GPC1, PCNA, IRF7, BRIP1, ULK1, and SPAG5 are selected.

[0073] In one embodiment, the selected at least two genes are MTUS1 and TTC21B.

[0074] In one embodiment, at least two genes include MTUS1 and TTC21B, and at least one additional gene is selected from MCM3, FOXM1, CUL4, GPC1, PCNA, IRF7, BRIP1, ULK1, and SPAG5. Preferably, at least one additional gene is MCM3 or FOXM1. Preferably, at least one gene is MCM3 and an additional FOXM1.

[0075] In one embodiment, at least two genes include MTUS1 and MCM3, and at least one additional gene is selected from TTC21B, FOXM1, CUL4, GPC1, PCNA, IRF7, BRIP1, ULK1, and SPAG5.

[0076] In one embodiment, at least two genes include TTC21B and MCM3, and at least one additional gene is selected from TTC21B, FOXM1, GPC1, CUL4, PCNA, MTUS1, IRF7, BRIP1, ULK1, and SPAG5.

[0077] In one embodiment, the at least two genes selected from TTC21B, MCM3, MTUS1, FOXM1, CUL4, GPC1, PCNA, IRF7, BRIP1, ULK1, and SPAG5 are MTUS1, TTC21B, MCM3, and FOXM1.

[0078] In one embodiment, the selected genes consist of MCM3, MTUS1, TTC21B, and FOXM1.

[0079] The term “consistent with essentially” should be understood to mean all eleven, ten, nine, eight, seven, six, five, four, three, or two genes selected from TTC21B, MCM3, MTUS1, FOXM1, CUL4A, GPC1, PCNA, IRF7, BRIP1, ULK1, and SPAG5.

[0080] In one embodiment, the cancer is selected from the group consisting of: multiple myeloma, prostate cancer, glioblastoma, lymphoma, fibrosarcoma; myxosarcoma; liposarcoma; chondrosarcoma; osteosarcoma; chordoma; angiosarcoma; endothelial sarcoma; lymphangiosarcoma; lymphangioendothelial sarcoma; synovial tumor; mesothelioma; Ewing's tumor. Tumors, leiomyosarcomas; rhabdomyosarcomas; colon cancer; pancreatic cancer; ovarian cancer; squamous cell carcinoma; basal cell carcinoma; adenocarcinoma; sweat gland carcinoma; sebaceous gland carcinoma; papillary carcinoma; papillary adenocarcinoma; cystadenocarcinoma; medullary carcinoma; bronchial protocarcinoma; renal cell carcinoma; hepatocellular carcinoma; cholangiocarcinoma; choriocarcinoma; seminoma; embryonal carcinoma; Wilms' tumor; cervical cancer; uterine cancer; testicular tumors; lung cancer; small cell lung cancer; bladder cancer; epithelial carcinoma; glioma; astrocytoma; medulloblastoma; craniopharyngioma; ependymoma; pineal tumor; hemangioblastoma; acoustic neuroma; oligodendroglioma; meningioma; melanoma; retinoblastoma; and leukemia. Appropriately, this cancer is epithelial carcinoma.

[0081] In one embodiment, the cancer is selected from the following precancerous lesions: Barrett's esophagus, Bowen's disease, familial adenomatous polyposis (FAP), lobular carcinoma in situ (LCIS), Lynch syndrome, MEN2, vaginal intraepithelial neoplasia (VAIN), and vulvar intraepithelial neoplasia (VIN). Precancerous lesions are included when diagnosing whether a subject is at risk of invasive cancer or cancer recurrence.

[0082] In one embodiment, recurrence is the development of a secondary tumor.

[0083] In one embodiment, the recurrence develops into a separate primary cancer unrelated to the sampled cancer.

[0084] In one embodiment of the present invention, a method for predicting the risk of recurrent prostate cancer in a patient is provided, the method comprising: wherein an individual has an increased risk of recurrent prostate cancer when the expression of at least two genes or proteins encoded by said genes is altered in a biological sample of a subject in which the expression of said at least two genes or proteins encoded by said genes is altered relative to the cancer being non-invasive or indolent.

[0085] In one embodiment, the therapy or treatment is a neoadjuvant therapy. In this specification, the term "neoadjuvant therapy" should be understood as meaning a treatment given prior to primary treatment to increase the chances of long-term survival. Primary treatment is typically surgery. Neoadjuvant therapy is typically selected from chemotherapy, hormone therapy, targeted therapy, radiation therapy, immunotherapy, or combinations thereof.

[0086] In one embodiment, the therapy is an adjunctive therapy. In this specification, the term "adjunctive therapy" should be understood to mean any treatment given after primary treatment to increase the chances of long-term survival. Primary treatment is typically surgery. Adjunctive therapies are typically selected from chemotherapy, hormone therapy, targeted therapy, radiation therapy, immunotherapy, or combinations thereof. Active surveillance can also be considered an adjunctive therapy.

[0087] In one embodiment, the therapy may be a combination of neoadjuvant therapy and adjuvant therapy.

[0088] In one embodiment, the therapy can be active monitoring, where the treatment plan involves closely observing an individual’s symptoms without administering any treatment unless test results indicate that the symptoms are worsening.

[0089] On the one hand, the treatment may be surgery (e.g., surgery to remove a primary or secondary tumor), or a combination of neoadjuvant therapy and surgery; or adjuvant therapy and surgery; or neoadjuvant therapy, surgery, and adjuvant therapy.

[0090] In one embodiment, a cancer patient may be eligible for neoadjuvant therapy to prevent cancer recurrence or progression.

[0091] In one embodiment, the cancer is prostate cancer.

[0092] On the one hand, adjuvant and neoadjuvant therapies are chemotherapy therapies. In one embodiment, the adjuvant and neoadjuvant therapies are selected from cyclophosphamide, methotrexate, 5-fluorouracil, gemcitabine, and doxorubicin. C 27 H 29 NO 11 ), paclitaxel ( C 47 H 51 NO 14 Capecitabine C 15 H 22FN3O6), docetaxel (FN3O6), C 43 H 53 NO 14 ), Cabazitaxel ( C 45 H 57 NO 14 ), mitoxantrone ( C 22 H 28 N4O6) and estramustine (3-(bis(2-chloroethyl)carbamate) estradiol ester; C 23 H 31 Cl2NO3).

[0093] On the one hand, the treatment options are to choose not to undergo surgery, adjuvant therapy, and / or neoadjuvant therapy.

[0094] In one embodiment of the invention, a method for treating cancer or precancerous conditions is provided, the method comprising the steps of: identifying an individual with increased cancer recurrence potential by measuring the expression of at least two genes selected from the group consisting of TTC21B, MCM3, MTUS1, FOXM1, CUL4A, GPC1, PCNA, IRF7, BRIP1, ULK1, and SPAG5 in a biological sample from an individual, wherein the individual has an increased risk of recurrent cancer when the normalized expression of the at least two genes or the proteins encoded by the genes changes relative to the normalized gene expression levels of the at least two genes from a biological sample obtained from a subject whose cancer is non-invasive or indolent; and

[0095] The individual was treated with an adjunctive therapy at a therapeutically effective dose.

[0096] In one embodiment, the individual is treated with a therapeutically effective dose of neoadjuvant therapy.

[0097] In one embodiment of the present invention, a method for treating cancer or precancerous conditions is provided, the method comprising the steps of: identifying an individual with increased cancer recurrence potential by measuring the expression of at least two genes selected from a group consisting of the following in a biological sample from an individual: TTC21B, MCM3, MTUS1, FOXM1, CUL4A, GPC1, PCNA, IRF7, BRIP1, ULK1, and SPAG5, wherein an increased risk of recurrent cancer is predicted when the normalized expression values ​​of the at least two genes or the combination of proteins encoded by said genes change relative to the combination of normalized expression values ​​of the at least two genes from a biological sample obtained from a subject with cancer; and

[0098] The individual was treated with a therapeutically effective dose of neoadjuvant therapy.

[0099] In one embodiment, the individual is treated with a therapeutically effective amount of adjunctive therapy.

[0100] In one embodiment, active monitoring is used to track individuals.

[0101] In one embodiment, the neoadjuvant therapy and adjuvant therapy are agents selected from, but not limited to, chemotherapeutic agents selected from: cyclophosphamide, methotrexate, 5-fluorouracil, gemcitabine, doxorubicin (…). C27H 29 NO 11 ), paclitaxel ( C 47 H 51 NO 14 ), capecitabine ( C 15 H 22 FN3O6), Dorcetase ( C 43 H 53 NO 14 ), Cabatha ( C 45 H 57 NO 14 ), Mitoxantrone ( C 22 H 28 N4O6) and estradiol ester (3-(bis(2-chloroethyl)carbamate); C 23 H 31 Cl2NO3) and luteinizing hormone-releasing (LHRH) agonists, these LHRH agonists are selected from leuprorelin ( C 59 H 84 N 16 O 12 ), Gosherelin ( C 59 H 84 N 18 O 14 ), Triptorelin ( C 64 H 82 N 18 O 13 ), Leuprolide Mesylate (C 60 H 88 N 16 O 15 S), Degarelk ( C 82 H 103ClN 18 O 16 ) and Rilugoli ( C 29 H 27 F2N7O5S).

[0102] This invention also relates to a method for treating an individual to prevent or suppress cancer recurrence, the method comprising the steps of: identifying a cancer patient at risk of recurrence using the method of this invention, and then treating the cancer patient with one or more agents to prevent or suppress cancer recurrence. Typically, the one or more agents comprise adjuvant or neoadjuvant therapy, or a combination of both.

[0103] In one embodiment, a system is provided for obtaining data from at least one test sample obtained from at least one individual, the system comprising: an assay module configured to receive at least one test sample and perform at least one test analysis on the test sample to determine the expression of at least three genes or proteins encoded by genes selected from the group consisting of: TTC21B, MCM3, MTUS1, FOXM1, CUL4A, GPC1, PCNA, IRF7, BRIP1, ULK1, and SPAG5; optionally, a storage system for storing expression data generated by the assay module; and a display module for displaying content based on data output from the assay module, wherein the content includes signals indicating a combination of normalized expression values ​​of the at least three genes or proteins encoded by those genes.

[0104] In one embodiment, at least three genes or proteins encoded by said genes are TTC21B, MTUS1, and MCM3.

[0105] In one embodiment, a method is provided for predicting the risk of cancer recurrence or progression in a patient and treating the patient with a therapy for preventing cancer recurrence, the method comprising the step of determining the expression of at least four genes selected from the group consisting of TTC21B, MCM3, MTUS1, FOXM1, CUL4A, GPC1, PCNA, IRF7, BRIP1, ULK1, and SPAG5 in a biological sample from the patient; wherein when the normalized expression values ​​of the at least four genes or a combination of proteins encoded by the genes change relative to the combination of normalized expression values ​​of the at least four genes obtained from a sample obtained from a cancer-free subject, the individual administers neoadjuvant or adjuvant therapy or a combination of both to the patient to prevent cancer recurrence or progression.

[0106] In one embodiment, at least four genes or proteins encoded by said genes are TTC21B, MTUS1, MCM3, and FOXM1.

[0107] As described herein, in subjects with cancer, and particularly in subjects with prostate cancer, the levels of one or more of TTC21B, MCM3, MTUS1, FOXM1, CUL4A, GPC1, PCNA, IRF7, BRIP1, ULK1, and SPAG5 can be altered or dysregulated. Therefore, in one aspect of any embodiment of the examples, this document describes a method for treating a subject with cancer or precancerous lesions, the method comprising administering local therapy (surgery, radiation therapy, or both), neoadjuvant therapy, adjuvant systemic therapy, or a combination thereof to a subject determined to have an altered combination of normalized expression values ​​of one or more of TTC21B, MCM3, MTUS1, FOXM1, CUL4A, GPC1, PCNA, IRF7, BRIP1, ULK1, and SPAG5 relative to a combination of normalized expression values ​​in a reference sample. In one aspect of any embodiment described herein, a method for treating a subject with prostate cancer in need is described, the method comprising: a) determining the levels of one or more of TTC21B, MCM3, MTUS1, FOXM1, CUL4A, GPC1, PCNA, IRF7, BRIP1, ULK1, and SPAG5 in a biological sample obtained from the subject; and b) administering the treatment to the subject if a combination of normalized expression values ​​of one or more of TTC21B, MCM3, MTUS1, FOXM1, CUL4A, GPC1, PCNA, IRF7, BRIP1, ULK1, and SPAG5 changes relative to a combination of normalized expression values ​​in a reference sample. The therapy may be selected from surgery and radiation therapy; and / or hormone therapy, chemotherapy, and / or immunotherapy, or combinations thereof. In one aspect, the therapy may be surgery (e.g., surgery to remove a primary or secondary tumor), or a combination of neoadjuvant therapy and surgery; or adjuvant therapy and surgery; or neoadjuvant therapy, surgery, and adjuvant therapy. Active surveillance may also be included.

[0108] On the one hand, adjuvant and neoadjuvant therapies are chemotherapy therapies. In one embodiment, the adjuvant and neoadjuvant therapies are selected from cyclophosphamide, methotrexate, 5-fluorouracil, gemcitabine, doxorubicin (… C27H29NO11), Paclitaxel ( C47H51NO14), capecitabine ( C15H22FN3O6), Dorcetase ( C43H53NO14), Cabazitax ( C45H57NO14), Mitoxantrone ( C22H28N4O6) and estradiol ester (3-(bis(2-chloroethyl)carbamate) estradiol ester; C23H31Cl2NO3).

[0109] In one embodiment, the selected genes are MTUS1 and TTC21B, MTUS1 and MCM3, MCM3 and TTC21B, or MCM3, MTUS1, TTC21B and FOXM1.

[0110] In some embodiments of any aspect, the method includes administration to a subject previously identified as having a combination of normalized expression values ​​of one or more of TTC21B, MCM3, MTUS1, FOXM1, CUL4A, GPC1, PCNA, IRF7, BRIP1, ULK1, and SPAG5, which is altered or dysregulated relative to a combination of normalized expression values ​​in a reference sample. In some embodiments of any aspect, this document describes a method for treating a subject with prostate cancer, the method comprising: a) first determining a combination of normalized expression values ​​of one or more of TTC21B, MCM3, MTUS1, FOXM1, CUL4A, GPC1, PCNA, IRF7, BRIP1, ULK1, and SPAG5 in a biological sample obtained from the subject; and b) if the combination of normalized expression values ​​of one or more of TTC21B, MCM3, MTUS1, FOXM1, CUL4A, GPC1, PCNA, IRF7, BRIP1, ULK1, and SPAG5 is altered or dysregulated relative to the combination of normalized expression values ​​in a reference sample, then administering to the subject a local therapy (surgery, radiation therapy, or both), neoadjuvant therapy, adjuvant systemic therapy, or a combination thereof.

[0111] In one embodiment, the selected genes are MTUS1 and TTC21B, MTUS1 and MCM3, MCM3 and TTC21B, or MCM3, MTUS1, TTC21B and FOXM1.

[0112] In one aspect of any of the embodiments described herein, a method for treating a subject with cancer or precancerous lesions is described, the method comprising: a) determining whether the subject has a combination of altered or dysregulated normalized expression values ​​of one or more of TTC21B, MCM3, MTUS1, FOXM1, CUL4A, GPC1, PCNA, IRF7, BRIP1, ULK1, and SPAG5; and b) if the combination of normalized expression values ​​of one or more of TTC21B, MCM3, MTUS1, FOXM1, CUL4A, GPC1, PCNA, IRF7, BRIP1, ULK1, and SPAG5 is altered or dysregulated relative to the combination of normalized expression values ​​in a reference sample, administering to the subject a local therapy (surgery, radiation therapy, or both), neoadjuvant therapy, adjuvant systemic therapy, or a combination thereof. In some embodiments of any aspect, the steps of determining whether a subject has a combination of altered or dysregulated normalized expression values ​​of one or more of TTC21B, MCM3, MTUS1, FOXM1, CUL4A, GPC1, PCNA, IRF7, BRIP1, ULK1, and SPAG5 may include (i) obtaining or having obtained a biological sample from the subject, and (ii) performing or having performed assays on the biological sample obtained from the subject to determine / measure the expression values ​​of one or more of TTC21B, MCM3, MTUS1, FOXM1, CUL4A, GPC1, PCNA, IRF7, BRIP1, ULK1, and SPAG5 in the subject. In some embodiments of any aspect, the step of determining whether a subject has altered or dysregulated expression values ​​of one or more of TTC21B, MCM3, MTUS1, FOXM1, CUL4A, GPC1, PCNA, IRF7, BRIP1, ULK1, and SPAG5 may include, or has been performed, on a biological sample obtained from the subject to determine / measure the expression values ​​of one or more of TTC21B, MCM3, MTUS1, FOXM1, CUL4A, GPC1, PCNA, IRF7, BRIP1, ULK1, and SPAG5 in the subject. In some embodiments of any aspect, the step of determining whether a subject has altered or dysregulated expression values ​​of one or more of TTC21B, MCM3, MTUS1, FOXM1, CUL4A, GPC1, PCNA, IRF7, BRIP1, ULK1, and SPAG5 may include ordering or requesting the determination of biological samples obtained from the subject to determine / measure the expression values ​​of one or more of TTC21B, MCM3, MTUS1, FOXM1, CUL4A, GPC1, PCNA, IRF7, BRIP1, ULK1, and SPAG5 in the subject.In some embodiments of any aspect, the step of determining whether a subject has altered or dysregulated expression values ​​of one or more of TTC21B, MCM3, MTUS1, FOXM1, CUL4A, GPC1, PCNA, IRF7, BRIP1, ULK1, and SPAG5 may include receiving assay results from a biological sample obtained from the subject to determine / measure expression values ​​of one or more of TTC21B, MCM3, MTUS1, FOXM1, CUL4A, GPC1, PCNA, IRF7, BRIP1, ULK1, and SPAG5 in the subject. In some embodiments of any aspect, the step of determining whether a subject has altered or dysregulated expression values ​​of one or more of TTC21B, MCM3, MTUS1, FOXM1, CUL4A, GPC1, PCNA, IRF7, BRIP1, ULK1, and SPAG5 may include receiving reports, results, or other methods for identifying a subject as having altered or dysregulated expression values ​​of one or more of TTC21B, MCM3, MTUS1, FOXM1, CUL4A, GPC1, PCNA, IRF7, BRIP1, ULK1, and SPAG5. In one embodiment, the selected genes are MTUS1 and TTC21B, MTUS1 and MCM3, MCM3 and TTC21B, or MCM3, MTUS1, TTC21B, and FOXM1.

[0113] In one aspect of any of the embodiments described herein, a method for treating a subject with prostate cancer in need is described, the method comprising: a) determining whether the subject has a combination of altered or dysregulated normalized expression values ​​of one or more of TTC21B, MCM3, MTUS1, FOXM1, CUL4A, GPC1, PCNA, IRF7, BRIP1, ULK1, and SPAG5; and b) if the combination of normalized expression values ​​of one or more of TTC21B, MCM3, MTUS1, FOXM1, CUL4A, GPC1, PCNA, IRF7, BRIP1, ULK1, and SPAG5 is altered or dysregulated relative to the combination of normalized expression values ​​in a reference sample, instructing or directing the subject to administer local therapy (surgery, radiation therapy, or both), neoadjuvant therapy, adjuvant systemic therapy, or a combination thereof. In some embodiments of any aspect, the steps of determining whether a subject has a combination of altered or dysregulated normalized expression values ​​of one or more of TTC21B, MCM3, MTUS1, FOXM1, CUL4A, GPC1, PCNA, IRF7, BRIP1, ULK1, and SPAG5 may include (i) obtaining or having obtained a biological sample from the subject, and (ii) performing or having performed assays on the biological sample obtained from the subject to determine / measure a combination of normalized expression values ​​of one or more of TTC21B, MCM3, MTUS1, FOXM1, CUL4A, GPC1, PCNA, IRF7, BRIP1, ULK1, and SPAG5 in the subject. In some embodiments of any aspect, the step of determining whether a subject has altered or dysregulated expression values ​​of one or more of TTC21B, MCM3, MTUS1, FOXM1, CUL4A, GPC1, PCNA, IRF7, BRIP1, ULK1, and SPAG5 may include, or has been, assaying a biological sample obtained from the subject to determine / measure a combination of normalized expression values ​​of one or more of TTC21B, MCM3, MTUS1, FOXM1, CUL4A, GPC1, PCNA, IRF7, BRIP1, ULK1, and SPAG5 in the subject.In some embodiments of any aspect, the step of determining whether a subject has altered or dysregulated expression levels of one or more of TTC21B, MCM3, MTUS1, FOXM1, CUL4A, GPC1, PCNA, IRF7, BRIP1, ULK1, and SPAG5 may include ordering or requesting a assay of a biological sample obtained from the subject to determine / measure a combination of normalized expression values ​​of one or more of TTC21B, MCM3, MTUS1, FOXM1, CUL4A, GPC1, PCNA, IRF7, BRIP1, ULK1, and SPAG5 in the subject. In some embodiments of any aspect, the step of instructing or directing a subject to administer a specific treatment may include providing a report of the assay results. In some embodiments of any aspect, the step of instructing or directing a subject to administer a specific treatment may include providing a report of the assay results and / or treatment recommendations based on the assay results. In one embodiment, the selected genes are MTUS1 and TTC21B, MTUS1 and MCM3, MCM3 and TTC21B, or MCM3, MTUS1, TTC21B and FOXM1.

[0114] In one aspect of any of the embodiments described herein, a method for determining whether a subject has cancer or requires cancer treatment is described, the method comprising: determining the expression of one or more of TTC21B, MCM3, MTUS1, FOXM1, CUL4A, GPC1, PCNA, IRF7, BRIP1, ULK1, and SPAG5 in a biological sample obtained from the subject, wherein a combination of normalized expression values ​​of one or more of TTC21B, MCM3, MTUS1, FOXM1, CUL4A, GPC1, PCNA, MTUS1, IRF7, BRIP1, ULK1, and SPAG5 is altered or dysregulated relative to a combination of normalized expression values ​​in a reference sample, thereby indicating a prediction that the subject has cancer or a prediction that the subject requires cancer treatment. In one aspect, the cancer is prostate cancer. In one embodiment, the selected genes are MTUS1 and TTC21B, MTUS1 and MCM3, MCM3 and TTC21B, or MCM3, MTUS1, TTC21B, and FOXM1.

[0115] On the one hand, there is a method for predicting an individual's risk of invasive or recurrent prostate cancer, which includes the step of measuring the expression of two genes, TTC21B and MTUS1, or proteins encoded by said genes, in a biological sample from the individual, wherein an increased risk of invasive or recurrent prostate cancer is predicted when the combination of normalized expression values ​​of the two genes or proteins encoded by said genes changes relative to the combination of normalized expression values ​​of the two genes or proteins encoded by said genes in a biological sample from a subject with non-invasive or non-recurrent prostate cancer.

[0116] On the one hand, there is a method for predicting the risk of prostate cancer recurrence or progression in an individual with prostate cancer after treatment, the method comprising: measuring the expression of two genes TTC21B and MTUS1 or proteins encoded by said genes in a biological sample obtained from the individual, wherein when the combination of normalized expression values ​​of the two genes or proteins encoded by said genes changes relative to the combination of normalized expression values ​​of the two genes from a biological sample obtained from a subject who has not experienced prostate cancer recurrence or progression after treatment, the risk of prostate cancer recurrence or progression in that individual is predicted to increase.

[0117] On the one hand, there is a method for determining the 5-year or 10-year survival rate of an individual diagnosed with prostate cancer, the method comprising: measuring the expression of two genes TTC21B and MTUS1 or proteins encoded by these genes in a biological sample from the individual, wherein a decrease in the individual's 5-year or 10-year survival rate is predicted when the combination of the normalized expression values ​​of the two genes or the proteins encoded by said genes changes relative to the combination of the normalized expression values ​​of the two genes in a biological sample obtained from a subject who survived prostate cancer 5 or 10 years after the diagnosis of prostate cancer.

[0118] On the one hand, there is a method for identifying prostate cancer patients suitable for treatment with a therapy for preventing recurrence or progression of prostate cancer, the method comprising: measuring the expression of two genes or proteins encoded by these genes TTC21B and MTUS1 in a biological sample from the individual, wherein the individual is predicted to be unsuitable for the therapy when the combination of normalized expression values ​​of the two genes or proteins encoded by said genes is the same as or increases after treatment with the therapy compared to the combination of normalized expression values ​​of the two genes or proteins encoded by said genes in a biological sample from the same individual before treatment with the therapy.

[0119] On the one hand, there is a method for monitoring the effectiveness of prostate cancer treatment in an individual with prostate cancer, the method comprising: measuring the expression of two genes or proteins encoded by these genes TTC21B and MTUS1 in a biological sample from the individual, wherein the treatment is ineffective and predicts an adverse outcome for the individual when the combination of normalized expression values ​​of the two genes or proteins encoded by said genes is the same as or increases after the administration of the treatment relative to the combination of normalized expression values ​​of the two genes in a biological sample from the same individual before the administration of the treatment.

[0120] On one hand, the method further includes the step of determining the expression of at least one gene or a protein encoded by said gene in the biological sample from the individual, the gene being selected from MCM3, FOXM1, CUL4A, GPC1, PCNA, IRF7, BRIP1, ULK1, and SPAG5. Preferably, the selected at least one gene is MCM3. On the other hand, the method further includes the step of determining the expression of at least two genes in the biological sample from the individual, the at least two genes being selected from CUL4A, MCM3, GPC1, PCNA, IRF7, BRIP1, ULK1, SPAG5, and FOXM1. Preferably, the at least two genes are MCM3 and FOXM1.

[0121] In some embodiments of any of these aspects, the measurement of target expression values ​​and / or the detection or presence of target expression values ​​(e.g., expression products (nucleic acids or polypeptides of a gene in the genes described herein) or mutations) may include transformation. As used herein, the terms “transforming” or “transformation” refer to the conversion of an object or substance, such as a biological sample, nucleic acid, or protein, into another substance. This transformation may be physical, biological, or chemical. Exemplary physical transformation includes, but is not limited to, pretreatment of biological samples, such as conversion from whole blood to serum by differential centrifugation. Biological / chemical transformation may involve the action of at least one enzyme and / or chemical reagent in the reaction. For example, a DNA sample may be digested into fragments by one or more restriction enzymes, or a foreign molecule may be ligated to a fragmented DNA sample using a ligase. In some embodiments of any of these aspects, the DNA sample may undergo enzymatic replication, such as by polymerase chain reaction (PCR).

[0122] The transformation, measurement, and / or detection of a target molecule (e.g., a gene, nucleotide, mRNA, or polypeptide) may include contacting a sample obtained from a subject with a reagent specific to the target (e.g., a detection reagent) (e.g., a target-specific reagent). In some embodiments of any aspect, the target-specific reagent is detectably labeled. In some embodiments of any aspect, the target-specific reagent may generate a detectable signal. In some embodiments of any aspect, the target-specific reagent generates a detectable signal in the presence of the target molecule.

[0123] Methods for measuring gene expression products are known to those skilled in the art. Such methods for measuring gene expression products (e.g., protein levels) include ELISA (enzyme-linked immunosorbent assay), Western blotting, immunoprecipitation, and immunofluorescence using detection reagents (such as antibodies or protein binders). Alternatively, peptides in a subject's body can be detected by introducing labeled anti-peptide antibodies and other types of detection reagents into the subject. For example, the antibody can be labeled with a detectable biomarker, the presence and localization of which in the subject's body can be detected using standard imaging techniques.

[0124] For example, the antibodies targeting various targets described herein are commercially available and can be used for the purposes of this invention to measure protein expression levels, such as anti-FOXM1 (Millibosigma / Merck, catalog number SAB1412254), anti-PTTG1 (Millibosigma / Merck, catalog number WH0009232M1), anti-ZNF367 (Millibosigma / Merck, catalog number HPA015785), anti-AGER (Millibosigma / Merck, catalog number AV41642), anti-CUL4A (Millibosigma / Merck, catalog number SAB1411512), anti-TTC21B (Millibosigma / Merck, catalog number HPA035495), and anti-MCM3. (Millibone Sigma / Merck Group, catalog number SAB1412669), Anti-GPC1 (Millibone Sigma / Merck Group, catalog number HPA030571), Anti-PCNA (Millibone Sigma / Merck Group, catalog number HPA030521), Anti-MTUS1 (Millibone Sigma / Merck Group, catalog number WH0057509M1), Anti-IRF7 (Millibone Sigma / Merck Group, catalog number PRS3941), Anti-BRIP1 (Millibone Sigma / Merck Group, catalog number HPA005474), Anti-ULK1 (Millibone Sigma / Merck Group, catalog number WH0008408M1), and Anti-SPAG5 (Millibone Sigma / Merck Group, catalog number HPA022479). Alternatively, since the amino acid sequences of the targets described herein are known and publicly available on the NCBI website, those skilled in the art can generate their own antibodies against these peptides of interest for the purposes of the methods described herein.

[0125] In some embodiments of any of these aspects, immunohistochemistry (“IHC”) and immunocytochemistry (“ICC”) techniques may be used. IHC applies immunochemistry to tissue sections, while ICC applies immunochemistry to cell or tissue imprints after they have undergone specific cytological preparation (e.g., liquid-based preparation). Immunochemistry is a series of techniques based on the use of antibodies, where antibodies are used to specifically target molecules inside or on the surface of cells. Antibodies typically contain a marker that undergoes a biochemical reaction upon encountering the target molecule, thereby experiencing a color change. In some cases, signal amplification may be integrated into a particular protocol that includes the application of a secondary antibody following the primary specific antibody, either through marker staining or a marker signal.

[0126] In some embodiments of any of these aspects, the assay may be Western blot analysis. Alternatively, proteins may be separated using a two-dimensional gel electrophoresis system. Two-dimensional gel electrophoresis is well known in the art and typically involves isoelectric focusing along a first dimension followed by SDS-PAGE electrophoresis along a second dimension. These methods also require large amounts of cellular material. Analysis of the 2D SDS-PAGE gel can be performed by measuring the intensity of protein spots on the gel, or it can be performed using immunoassay. In other embodiments, protein samples are analyzed by mass spectrometry.

[0127] Immunoassays can be used in conjunction with the methods and assays described herein and include, for example, competitive and non-competitive assay systems using techniques such as Western blotting, radioimmunoassay (RIA), ELISA (enzyme-linked immunosorbent assay), sandwich immunoassay, immunoprecipitation assay, immunodiffusion assay, agglutination assay (e.g., latex agglutination), complement fixation assay, immunoradioassay, fluorescence immunoassay (e.g., FIA (fluorescence-linked immunoassay)), chemiluminescent immunoassay (CLIA), electrochemiluminescent immunoassay (ECLIA, count immunoassay (CIA)), lateral flow assay or immunoassay (LFIA), magnetic immunoassay (MIA), and protein A immunoassay. Methods for performing such assays are known in the art, provided suitable antibody reagents are available. In some embodiments of any aspect, the immunoassay may be a quantitative or semi-quantitative immunoassay.

[0128] Immunoassays are biochemical assays that use the interaction of one or more antibodies with their antigens to measure the concentration of a substance in a biological sample (typically a fluid sample such as blood or serum). The assays take full advantage of the highly specific binding of antibodies to their antigens. For the methods and assays described herein, the specific binding of a target peptide to a corresponding protein or protein fragment, or an isolated peptide, or a fusion protein as described herein, occurs in the immunoassay to form a target protein / peptide complex. The complex is then detected by a variety of methods known in the art. Immunoassays also frequently involve the use of detection antibodies.

[0129] Enzyme-linked immunosorbent assay (ELISA), also known as enzyme immunoassay or EIA, is a biochemical technique primarily used in immunology to detect the presence of antibodies or antigens in a sample. ELISA has been used as a diagnostic tool in medicine and phytopathology, as well as for quality control checks in various industries.

[0130] In one embodiment, an ELISA can also be performed involving at least one antibody specific to a desired antigen (e.g., any of the targets described herein). A known amount of sample and / or antigen is immobilized on a solid support (typically a polystyrene microtiter plate). Immobilization can be nonspecific (e.g., by adsorption to a surface) or specific (e.g., another antibody immobilized on the surface to capture the antigen or a primary antibody). After antigen immobilization, a detection antibody is added, thereby forming a complex with the antigen. The detection antibody can be covalently linked to an enzyme or can be detected by a secondary antibody linked to the enzyme via bioconjugation. Between each step, the plate is typically washed with a mild detergent solution to remove any nonspecifically bound proteins or antibodies. After the final washing step, the plate is colored by adding an enzymatic substrate to produce a visible signal indicating the amount of antigen in the sample. Older ELISAs utilized chromogenic substrates, although newer assays use much more sensitive fluorescent substrates.

[0131] In another embodiment, a competitive ELISA is used. Purified antibodies against a target peptide or fragment thereof are coated onto the solid phase of a multi-well plate, i.e., conjugated to a solid surface. A second batch of purified antibodies, unconjugated on any solid support, is also required. These unconjugated purified antibodies are labeled for detection purposes, for example, with horseradish peroxidase to generate a detectable signal. A sample from a subject (e.g., a blood sample) is mixed with a known amount of the desired antigen (e.g., a known volume or concentration of a sample containing the target peptide) and the horseradish peroxidase-conjugated antibody, and the mixture is then added to the coated wells to form a competitive combination. After incubation, if the peptide level in the sample is high, a labeled antibody-antigen complex will form. This complex is free in solution and can be washed away. Washing the wells removes the complex. The wells are then incubated with a TMB (3,3',5,5'-tetramethylphenylene) chromogenic substrate to localize the horseradish peroxidase-conjugated antibody in the wells. If the target peptide level in the sample is high, there will be no color change or a very small color change. If there are few or no target peptides in the sample, a different complex is formed—an antibody-target peptide complex bound to the solid-phase support. This complex is immobilized on the plate and is not washed away during the washing step. Subsequent incubation with TMB will produce a significant color change. This type of competitive ELSA assay is specific, sensitive, reproducible, and easy to operate.

[0132] Other different forms of ELISA are well known to those skilled in the art. Standard techniques for ELISA known in the art are described in "Methods in Immunodiagnosis," 2nd edition, edited by Rose and Bigazzi, John Wiley & Sons, 1980; and Oellerich, M., 1984, J. Clin. Chem. Clin. Biochem. 22: 895-904. These references are hereby incorporated in their entirety by reference.

[0133] In one embodiment, the level of a peptide in a sample can be detected by a lateral flow immunoassay (LFIA) (also known as an immunochromatographic assay or strip test). An LFIA is a simple device designed to detect the presence (or absence) of an antigen (e.g., a peptide) in a fluid sample. Many LFIA tests are currently available for medical diagnostics, for home testing, point-of-care testing, or laboratory use. An LFIA test is a form of immunoassay in which a test sample flows along a solid substrate via capillary action. After the sample is applied to a test strip, it encounters a colored reagent (typically including an antibody specific to the target antigen) that binds to particles, which mix with the sample and pass through the substrate, thus encountering a line or region pretreated with another antibody or antigen. Depending on the level of the target peptide present in the sample, the colored reagent can be captured and bound at the test line or region. An LFIA is essentially an immunoassay adapted for operation along a single axis to accommodate either a test strip or a strip format. Strip tests are very versatile and can be readily modified by those skilled in the art to detect large amounts of antigen from fluid samples such as urine, blood, water, and / or homogenized tissue samples. The LFIA strip test, also known as the dip test, derives its name from the literal meaning of "immersing" the test strip into the liquid sample being tested. LFIA strip tests are easy to use, require minimal training, and can be readily used in the field as part of point-of-care testing (POCT) diagnostics. LFIA tests can be performed as competitive or sandwich assays. A sandwich LFIA is similar to a sandwich ELISA. The sample first encounters colored particles labeled with antibodies against the target antigen. The test line will also contain antibodies against the same target, although these may bind to different epitopes on the antigen. In positive samples, the test line will appear as a colored band. In some embodiments of any aspect, the lateral flow immunoassay can be a double-antibody sandwich assay, a competitive assay, a quantitative assay, or a variant thereof. A competitive LFIA is similar to a competitive ELISA. The sample first encounters colored particles labeled with the target antigen or analogue. The test line contains antibodies against the target / analyte. Unlabeled antigens in the sample will block binding sites on the antibodies, thus preventing the uptake of the colored particles. In negative samples, the test line will appear as a colored band. There are several variations of lateral flow techniques. Multiple capture regions can also be applied to create multiple tests.

[0134] In the context of immunoassays of numerous antigen biomarkers, the use of "dip sticks" or LFIA test strips and other solid-phase carriers has been described in the art. U.S. Patent Nos. 4,943,522; 6,485,982; 6,187,598; 5,770,460; 5,622,871; 6,565,808; U.S. Patent Application Serial No. 10 / 278,676; U.S. Serial No. 09 / 579,673; and U.S. Serial No. 10 / 717,082 (these documents are incorporated herein by reference in their entirety) are non-limiting examples of such lateral flow assay devices. Examples of patents describing the use of "dip stick" technology for the detection of soluble antigens by immunochemical assay include, but are not limited to, U.S. Patent Nos. 4,444,880; 4,305,924; and 4,135,884; the entirety of which are incorporated herein by reference. These three patents broadly describe the apparatus and methods for a first component immobilized on a solid surface of a "dipping rod," which is then exposed to a solution containing a soluble antigen bound to the component immobilized on the "dipping rod," prior to the detection of the component-antigen complex on the rod. Those skilled in the art will be able to modify the teachings of this "dipping rod" technique to detect peptides using the antibody reagents described herein.

[0135] Other techniques can be used to detect the level of peptides in a sample. One such technique is dot blotting, an improvement on protein blotting (Towbin et al., Proc. Nat. Acad. Sci. 76:4350 (1979)). In protein blotting, a peptide or fragment thereof can be dissociated with a detergent and heated, separated on an SDS-PAGE gel, and then transferred to a solid support such as a nitrocellulose or PVDF membrane. The membrane is incubated with an antibody reagent specific to the target peptide or fragment thereof. The membrane is then washed to remove unbound proteins and proteins with nonspecific binding. Detectable labeled enzyme-linked secondary antibodies or detection antibodies can then be used to detect and assess the amount of peptide in the tested sample. Dot blotting immobilizes a protein sample on a defined area of ​​a support, which is then detected by antibodies and labeled secondary antibodies, as in protein blotting. The signal intensity from a detectable label of either format corresponds to the amount of enzyme present, and therefore to the amount of peptide. The level can be quantified, for example, by densitometric assay.

[0136] In some embodiments of any aspect, by way of non-limiting examples, the level of the target can be measured by: Western blotting; immunoprecipitation; enzyme-linked immunosorbent assay (ELISA); radioimmunoassay (RIA); sandwich assay; fluorescence in situ hybridization (FISH); immunohistochemical staining; radioimmunoassay; immunofluorescence; colorimetric assay; mass spectrometry and / or immunoelectrophoresis.

[0137] In some embodiments, the gene expression products described herein can be determined by measuring the expression level of the messenger RNA (mRNA) of the genes described herein. Such molecules can be isolated, derived, or amplified from biological samples (such as blood samples). Techniques for detecting mRNA expression are known to those skilled in the art and may include, but are not limited to, PCR procedures, RT-PCR, quantitative RT-PCR, Northern blotting analysis, differential gene expression, RNase protection assays, microarray-based analysis, next-generation sequencing, hybridization methods, etc.

[0138] Generally, a PCR procedure describes a method for gene amplification that includes (i) sequence-specific hybridization of primers with a specific gene or sequence in a nucleic acid sample or library, (ii) subsequent amplification involving multiple rounds of annealing, extension, and denaturation using a thermostable DNA polymerase, and (iii) screening for bands of the correct size in the PCR product. The primers used are oligonucleotides of sufficient length and appropriate sequence to provide the initiation of polymerization; that is, each primer is specifically designed to be complementary to one strand of the genomic locus to be amplified. In alternative embodiments, the mRNA level of the gene expression product described herein can be determined by reverse transcription (RT) PCR and quantitative RT-PCR (QRT-PCR) or real-time PCR methods. RT-PCR and QRT-PCR methods are well known in the art.

[0139] In some embodiments of any of these aspects, the level of mRNA can be measured using quantitative sequencing techniques (e.g., quantitative next-generation sequencing). Sequencing methods for nucleic acid sequences are well known in the art. Briefly, a sample obtained from a subject can be contacted with one or more primers that specifically hybridize with a single-stranded nucleic acid sequence flanking a target gene sequence, and a complementary strand can be synthesized. In some next-generation techniques, an adaptor (double-stranded or single-stranded) is attached to a nucleic acid molecule in the sample, and synthesis begins from the adaptor or an adaptor-compatible primer. In some third-generation techniques, the sequence can be determined, for example, by determining the location and pattern of probe hybridization or by measuring one or more properties of a single molecule as it passes through a sensor (e.g., modulation of the electric field as the nucleic acid molecule passes through a nanopore). Exemplary sequencing methods include, but are not limited to, Sanger sequencing, dideoxy chain termination, high-throughput sequencing, next-generation sequencing, 454 sequencing, SOLiD sequencing, polony sequencing, Illumina sequencing, ion-fluidic sequencing, hybridization sequencing, nanopore sequencing, Helioscope sequencing, single-molecule real-time sequencing, RNAP sequencing, etc. The methods and protocols used to perform these sequencing methods are known in the art; see, for example, “Next Generation Genome Sequencing,” edited by Michal Janitz and Wiley-VCH; “High-Throughput Next Generation Sequencing,” edited by Kwon and Ricke, Humanna Press, 2011; and Sambrook et al., “Molecular Cloning: A Laboratory Manual” (4th edition), Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA (2012); these references are incorporated herein by reference in their entirety.

[0140] The nucleic acid sequences of the genes described in this article have been assigned NCBI accession numbers for different species (e.g., human, mouse, and rat). For example, the nucleic acid sequences (e.g., SEQ ID NO: 1-11) and mRNA of human TTC21B, MCM3, MTUS1, FOXM1, CUL4A, GPC1, PCNA, IRF7, BRIP1, ULK1, and SPAG5 are known. Similarly, the NCBI accession numbers for cancer reference markers ALAS1, MMADHC, PPP2CA, TFRC, and CLTC are also known (e.g., SEQ ID NO: 12-16). Therefore, technicians can design suitable primers based on known sequences used to determine the nucleic acid or mRNA levels of the corresponding genes.

[0141] Nucleic acid and ribonucleic acid (RNA) molecules can be isolated from a specific biological sample using any of the many procedures well known in the art, with the specific isolation procedure chosen being suitable for the specific biological sample. For example, freeze-thaw and alkaline lysis procedures can be used to obtain nucleic acid molecules from solid materials; heat and alkaline lysis procedures can be used to obtain nucleic acid molecules from urine; and proteinase K extraction can be used to obtain nucleic acids from blood (Roiff, A et al., PCR: Clinical Diagnostics and Research, Springer (1994)).

[0142] In some embodiments of any aspect, one or more reagents described herein (e.g., antibody reagents and / or nucleic acid probes) may include detectable labels and / or include the ability to generate detectable signals (e.g., a reaction that catalyzes the conversion of a compound into a detectable product). Detectable labels may include, for example, light-absorbing dyes, fluorescent dyes, or radioactive labels. Detectable labels, methods for detecting such detectable labels, and methods for incorporating such detectable labels into reagents (e.g., antibodies and nucleic acid probes) are well known in the art.

[0143] In some embodiments of any aspect, the detectable label may comprise a label detectable by spectroscopic, photochemical, biochemical, immunochemical, electromagnetic, radiochemical, or chemical methods (such as fluorescence, chemiluminescence, or any other suitable method). The detectable label used in the methods described herein may be a primary label (where the label includes or produces a directly detectable portion) or a secondary label (where the detectable label binds to another portion to produce a detectable signal, e.g., as is common in immunological labeling using secondary and tertiary antibodies). The detectable label may be conjugated to a reagent by covalent or non-covalent methods. Alternatively, the detectable label may be conjugated as by directly labeling a molecule that binds to the reagent via a ligand-receptor pair arrangement or other such specifically recognizing molecules. The detectable label may comprise, but is not limited to, radioisotopes, bioluminescent compounds, chromophores, antibodies, chemiluminescent compounds, fluorescent compounds, metal chelates, and enzymes.

[0144] In other embodiments, the detection reagent is labeled with a fluorescent compound. The presence of the fluorescently labeled reagent can be detected by fluorescence when exposed to light of an appropriate wavelength. In some embodiments of any aspect, the detectable label may be a fluorescent dye molecule or fluorophore, which includes, but is not limited to, fluorescein, phycoerythrin, phycocyanin, phthalaldehyde, fluorescein, and Cy3. TM Cy5 TM Allophycocyanin, Texas Red, peridinin chlorophyll, anthocyanins, and tandem conjugates (such as phycoerythrin-Cy5) TM ), green fluorescent protein, rhodamine, fluorescein isothiocyanate (FITC), and Oregon Green TM Rhodamine and its derivatives (e.g., Texas Red and tetrarhodamine isothiocyanate (TRITC)), biotin, phycoerythrin, AMCA, CyDyes TM6-Carboxyfluorescein (usually abbreviated as FAM and F), 6-carboxy-2',4',7',4,7-hexachlorofluorescein (HEX), 6-carboxy-4',5'-dichloro-2',7'-dimethoxyfluorescein (JOE or J), N,N,N',N'-tetramethyl-6-carboxyrhodamine (TAMRA or T), 6-carboxy-X-rhodamine (ROX or R), 5-carboxyrhodamine-6G (R6G5 or G5), 6-carboxyrhodamine-6G (R6G6 or G6), and rhodamine 110; anthocyanin dyes (e.g., Cy3, Cy5, and Cy7 dyes); coumarins (e.g., umbelliferone); benzamide dyes (e.g., Hoechst... 33258); phenanthridine dyes (e.g., Texas Red); ethidium dyes; acridine dyes; carbazole dyes; phenoxazine dyes; porphyrin dyes; polymethystyl dyes (e.g., cyanide dyes, such as Cy3, Cy5, etc.); BODIPY dyes and quinoline dyes. In some embodiments of any aspect, the detectable marker may be a radioactive marker, including but not limited to... 3 H, 125 I, 35 S, 14 C. 32 P and 33 P. In some embodiments of any aspect, the detectable label may be an enzyme, including but not limited to horseradish peroxidase and alkaline phosphatase. Enzymatic labeling may produce, for example, chemiluminescent, color, or fluorescent signals. Enzymes intended for detectably labeling antibody reagents include, but are not limited to, malate dehydrogenase, staphylococcal nuclease, δ-V-steroid isomerase, yeast alcohol dehydrogenase, α-glycerophosphate dehydrogenase, triose phosphate isomerase, horseradish peroxidase, alkaline phosphatase, asparaginase, glucose oxidase, β-galactosidase, ribonuclease, urease, catalase, glucose-VI-phosphate dehydrogenase, glucosylamylase, and acetylcholinesterase. In some embodiments of any aspect, the detectable label is a chemiluminescent label, including but not limited to lucigenin, luminol, fluorescein, isoluminol, thermoacridone ester, imidazole, acridine salt, and oxalate ester. In some embodiments of any aspect, the detectable marker may be a spectral colorimetric marker comprising, but not limited to, colloidal gold or colored glass or plastic (e.g., polystyrene, polypropylene, and latex) beads.

[0145] In some embodiments of any of these aspects, the assay reagents may also be labeled with a detectable tag (such as c-Myc, HA, VSV-G, HSV, FLAG, V5, HIS, or biotin). Other assay systems may also be used, such as the biotin-streptavidin system. In this system, an antibody that reacts immunologically with the biomarker of interest (i.e., is specific) is biotinylated. The amount of biotinylated antibody binding to the biomarker is determined using a streptavidin-peroxidase conjugate and a chromogenic substrate. Such streptavidin peroxidase assay kits are commercially available, for example, from DAKO Corporation (Carpinteria, CA). The reagents may also use fluorescent emitting metals (such as...) 152 The labeling of Eu or other metals of the lanthanides can be detectably performed. These metals can be linked to reagents using metal chelating groups such as diethylenetriaminepentaacetic acid (DTPA) or ethylenediaminetetraacetic acid (EDTA).

[0146] An expression value or risk score lower than a reference expression value or reference risk score can be an expression value that is at least about 10%, at least about 20%, at least about 50%, at least about 60%, at least about 80%, at least about 90%, or less lower than a reference expression value or reference risk score. In some embodiments of any aspect, an expression value or risk score lower than a reference expression value or reference risk score can be an expression value or risk score that is statistically significantly lower than a reference expression value or reference risk score.

[0147] An expression value or risk score greater than a reference expression value or reference risk score can be at least about 10%, at least about 20%, at least about 50%, at least about 60%, at least about 80%, at least about 90%, at least about 100%, at least about 200%, at least about 300%, at least about 500%, or more, or less than the reference expression value or reference risk score. In some embodiments of any aspect, an expression value or risk score greater than or less than the reference expression value or reference risk score can be an expression value or risk score that is statistically significantly greater than the reference expression value or reference risk score.

[0148] In some embodiments of any aspect, the reference may be the expression value of the target molecule in a population of subjects who do not have or have not been diagnosed with and / or do not show signs or symptoms of cancer. In some embodiments of any aspect, the reference may also be the expression value of the target molecule in a control sample, a pooled sample of control individuals, or a numerical value or range of values ​​based on that expression value. In some embodiments of any aspect, the reference may be the expression value of the target molecule in a sample obtained from the same subject at an earlier time point; for example, the methods described herein can be used to determine whether a subject's sensitivity or response to a given therapy has changed over time.

[0149] In some respects, risk scores can be used when using combinations of gene expression levels to determine outcomes or the direction of action for subjects who have cancer, have been diagnosed with cancer, are suspected of having cancer, or are undergoing cancer treatment. The “direction of action” of gene increases or decreases in cancer patients is based on a single-gene model of adverse pathological (AP) outcomes. However, when multiple genes (two or more) are present in the marker, the direction of action of gene expression levels can be altered (also known as altered expression). A more general description of the accurate and implicit direction of action for any combination of genes is the use of risk scores (associated with altered expression). To calculate a risk score, a weighted linear combination of the normalized expression values ​​of multiple genes is used to calculate the risk score for any combination of genes. Risk scores estimate, for example, the probability of invasive cancer; a higher risk score indicates a greater likelihood of developing invasive cancer, and vice versa. This is also applicable to determining the efficacy of treatment, or whether treatment is appropriate, or whether the prognosis for a cancer patient is poor or good. For example, the terms "gene combination" or "risk score" should be understood as referring to a linear combination or weighted sum of the normalized expression of the genes in question. A higher risk score indicates a higher risk of aggressive or recurrent cancer, and a worse outcome, resulting in lower 5- or 10-year survival rates and less effective treatment. Conversely, a lower risk score indicates a lower risk of aggressive or recurrent cancer, and a better outcome, resulting in higher 5- and / or 10-year survival rates and more effective treatment. The risk score for N gene combinations is calculated as follows:

[0150] Risk score = W1 x G1 + W2 x G2 + ... + W N x G N (Equation 1)

[0151] in,

[0152] W = a gene-specific real constant that can be positive or negative.

[0153] G = Gene-specific normalized expression value

[0154] In some embodiments of any aspect, the expression values ​​of the expression products of no more than 200 other genes are measured. In some embodiments of any aspect, the expression values ​​of the expression products of no more than 100 other genes are measured. In some embodiments of any aspect, the expression values ​​of the expression products of no more than 20 other genes are measured. In some embodiments of any aspect, the expression values ​​of the expression products of no more than 15 other genes are measured. In some embodiments of any aspect, the expression values ​​of the expression products of no more than 10 other genes are measured.

[0155] In some embodiments of the foregoing aspects, the expression value of a given gene may be normalized relative to the expression values ​​of one or more reference genes or reference proteins.

[0156] In some embodiments, the reference value may be an expression value from a sample / subject with similar cell type, sample type, sample processing, and / or from a sample obtained from a subject with similar age, sex, and other demographic parameters to the expression values ​​of one or more of TTC21B, MCM3, MTUS1, FOXM1, CUL4A, GPC1, PCNA, IRF7, BRIP1, ULK1, and SPAG5, FOXM1 to the expression values ​​to be determined. In some embodiments, the test sample and the control reference sample are of the same type, i.e., obtained from the same biological source, and comprise the same composition, e.g., the same number and cell type of cells.

[0157] Test samples can be obtained by removing samples from the subject, but they can also be obtained by using previously separated samples (e.g., separated at a previous time point and by the same person or another person).

[0158] In some embodiments of any aspect, the test sample may be an untreated test sample. As used herein, the phrase "untreated test sample" means a test sample that has not undergone any sample pretreatment other than dilution and / or suspension in solution. Exemplary methods for treating test samples include, but are not limited to, centrifugation, filtration, sonication, homogenization, heating, freezing and thawing, and combinations thereof. In some embodiments of any aspect, the test sample may be a frozen test sample (e.g., frozen tissue). Frozen samples may be thawed before being subjected to the methods, assays, and systems described herein. After thawing, frozen samples may be centrifuged before undergoing the methods, assays, and systems described herein. In some embodiments of any aspect, the test sample is a clarified test sample, for example, by centrifugation and collection of a supernatant comprising a clarified test sample. In some embodiments of any aspect, the test sample may be a pretreated test sample, for example, a supernatant or filtrate produced by treatment selected from the group consisting of centrifugation, filtration, thawing, purification, and any combination thereof. In some embodiments of any aspect, the test sample may be treated with chemical and / or biological reagents. During processing, chemical and / or biological reagents may be employed to protect and / or maintain the stability of the sample, including the biomolecules contained therein (e.g., nucleic acids and proteins). An exemplary reagent is a protease inhibitor, which is commonly used to protect or maintain the stability of proteins during processing. Those skilled in the art will recognize methods and procedures suitable for biological sample pretreatment required for determining the level of expressed products as described herein.

[0159] In some embodiments of any aspect, the methods, assays, and systems described herein may further include the step of obtaining or having obtained a test sample from a subject. In some embodiments of any aspect, the subject may be a human subject. In some embodiments of any aspect, the subject may be a subject requiring treatment (e.g., having or being diagnosed with prostate cancer), or a subject at risk of or with an increased risk of developing prostate cancer as described elsewhere herein.

[0160] In some embodiments of any aspect, the sample obtained from the subject may be a biopsy sample. In some embodiments of any aspect, the sample obtained from the subject may be a blood or serum sample. Attached Figure Description

[0161] The invention will be more clearly understood with reference to the accompanying drawings, in which:

[0162] Figure 1Four figures (A)-(D) are shown, illustrating the prognostic performance of adverse pathological (AP) outcomes for 512 combinations of 2-11 genes, including MTUS1 and TTC21B, and one or more of the other nine genes. Figure 1 (A)). The combination of the two genes MTUS1 and TTC21B is highlighted as a red dot, indicating a significant prognosis (p = 0.0013) and significantly increasing the prognostic value for the following: baseline clinical information ( Figure 1 (B) Optimal weighted biopsy information consisting of Gleason score, clinical stage, and PSA (p = 0.0101), CAPRA risk score ( Figure 1 (C), clinical information consisting of the Prostate Cancer Risk Assessment (CAPRA) score from 0 to 10 (p = 0.0082) and EAU risk category ( Figure 1 (D), clinical information comprised of low, intermediate, or high-risk categories according to the European Association of Urology (p = 0.0028). AP was determined by the likelihood ratio statistic (LRS) of cross-validation using the area under the ROC curve (AUC). Larger LRS and AUC values ​​indicate better prognostic performance; therefore, points closer to the upper right corner of each plot represent gene combinations with better prognostic performance. Baseline clinical information is shown by blue dots, single genes by black dots, two-gene combinations of MTUS1 and TTC21B by red dots, and four-gene combinations of MTUS1, TTC21B, MCM3, and FOXM1 by green dots. Values ​​to the right of the dashed line are statistically significant (LRS p < 0.05).

[0163] Figure 2 The prognostic performance of biochemical relapse was presented for 512 gene combinations. Each point represents one unique gene combination out of the 512. Red dots represent the 2-gene combination MTUS1 and TTC21B. Green dots represent the 4-gene combination FOXM1, MCM3, MTUS1, and TTC21B. Prognostic performance was measured by likelihood ratio statistic (LRS) and C-index from cross-validation analysis. Larger LRS and C-index values ​​indicated better prognostic performance; therefore, points closer to the upper right corner of the graph represent gene combinations with better prognostic performance. Values ​​to the right of the vertical dashed line were considered statistically significant, with an LRS p-value < 0.05.

[0164] Figure 3A graph showing the prognostic performance of adverse pathological and biochemical relapses for 512 gene combinations is presented. Two-gene combinations MTUS1 and TTC21B are indicated by red dots, and four-gene combinations FOXM1, MCM3, MTUS1, and TTC21B are indicated by green dots. Prognostic performance was measured by the area under the ROC curve (AUC) and the C-index from cross-validation analysis. Larger AUC and C-index values ​​indicate better prognostic performance; therefore, dots closer to the upper right corner of the graph represent gene combinations with better prognostic performance. Detailed Implementation

[0165] definition

[0166] In this specification, the term "cancer sample" should be understood to mean tumor cells, tumor tissue, or other biological material derived from a tumor, such as conditioned medium.

[0167] As used herein, the terms “sample,” “biological sample,” or “test sample” mean a sample taken from or isolated from a biological organism, such as a blood or plasma sample from a subject. In some embodiments of any aspect, the invention covers several instances of biological samples. In some embodiments of any aspect, a biological sample is a cell, or tissue, or peripheral blood, or body fluid. Exemplary biological samples include, but are not limited to, biopsies, biological samples (e.g., prostate cancer samples, colorectal cancer samples, lung cancer samples, head and neck cancer samples, samples for examining precancerous lesions (e.g., Barrett esophagoscopy samples), etc.), biological fluid samples; blood; blood derivatives; serum; plasma; urine; semen; mucus; tissue biopsy; organ biopsy; synovial fluid; bile; cerebrospinal fluid; mucosal secretions; effusions; sweat; saliva; and / or tissue samples, etc. The term also includes mixtures of the above samples. The term “test sample” also includes untreated or pretreated (or pre-processed) biological samples. In some embodiments of any aspect, the test sample may include cells from the subject. In some embodiments of any aspect, the test sample may be a urine sample or tissue sample from the subject, wherein the urine sample or the tissue sample may contain tumor cells. In addition to the definitions summarized above, the sample may also contain non-tumor tissue, precancerous tissue, conditioned medium, or formalin-fixed paraffin-embedded (FFPE) tumor or non-tumor tissue.

[0168] In this specification, the term "positive expression," when applied to a gene or the protein encoded by that gene, should be understood to mean an increased expression level (expression value) of that gene or the protein encoded by that gene that is higher than the mean expression level of the same gene or the protein encoded by that same gene found in a cohort of matched control individuals ("control group") with cancer. The cohort of matched individuals may consist of individuals who have not experienced cancer recurrence after surgical removal of the cancer. Regarding controls, it is common practice among those skilled in the art to use 'standard' controls, for example, for immunohistochemistry (IHC), one or more cell lines are known for the expression level of the biomarker, or for qPCR (quantitative polymerase chain reaction), a similar standard control or pool of many samples is known.

[0169] In this specification, the terms "dysregulated expression" or "altered expression" should be understood as meaning a normalized expression value that is greater than or less than a normalized expression value that is present in a cohort of matched individuals with cancer (who have not had cancer after surgical removal or non-surgical treatment and have not had cancer recurrence) or in a cohort of matched individuals without cancer or who have never been diagnosed with cancer. In other words, the term should be understood as meaning a risk score calculated from the expression of a gene or gene combination that is greater than or less than a risk score calculated from the expression of the same gene or gene combination in individuals with cancer (who have not had cancer after surgical removal or non-surgical treatment and have not had cancer recurrence) or in a cohort of matched individuals without cancer or who have never been diagnosed with cancer.

[0170] In this specification, the term "normalized gene expression" should be understood as referring to the expression value of a gene regulated by the expression value of a control or reference gene. In this case, the reference genes used are PPP2CA, CLTC, ALAS1, MMADHC, and / or TFRC (SEQ ID NO: 12-16). The normalized gene expression of gene G (or a combination of genes G) is calculated as follows:

[0171] Normalized gene expression = (average RT-qPCR quantitative cycle value of the set of reference genes) - (RT-qPCR quantitative cycle value of gene G (or combination of genes G)) (Equation 2)

[0172] The term "normal expression," when applied to genes or proteins, should be understood to mean that the expression level of that gene (or the protein encoded by that gene) is equal to the expression level of the same gene or the protein encoded by that same gene found in a cohort of matched control individuals with cancer. The cohort of matched individuals may consist of individuals who have not experienced cancer recurrence after surgical removal of the cancer.

[0173] The methods used to set thresholds differ for microarray analysis, qRT-PCR analysis, and protein expression. For microarrays, the threshold is relative (samples are divided into three equal groups, so the threshold is dataset-dependent), and for qPCR and protein expression, the threshold is set to a specific point. For RNA (microarrays), 'low', 'intermediate', and 'high' expression levels refer to expression values ​​falling within the lower, middle, or upper third of the expression range; or alternatively, 'low' and 'high' expression can refer to expression values ​​falling within the lower or upper half of the expression range. For qRT-PCR and protein expression levels, specific thresholds have been set, but generally, the term "dysregulation" refers to tumors with expression values ​​higher or lower than the set value within the expression range. The terms "intermediate" and "normal" refer to tumors with expression values ​​falling within the set value within the expression range. For example, firstly, the gene expression values ​​(or protein levels) of at least two of the 11 genes are normalized against the gene expression values ​​(or protein levels) of reference genes PPP2CA, CLTC, ALAS1, MMADHC, and / or TFRC in the same biological sample, and a Ct value is obtained for each of the 16 genes. The average expression level of all reference genes is calculated, and the average RefCt value is obtained. The normalized expression level (ΔCTgene1) of each of the 11 genes is calculated by subtracting the expression level of the gene of interest (Ctgene1) from the average RefCt value (average RefCt - Ctgene1 = ΔCTgene1). Then, the ΔCT of at least two of the 11 genes is obtained, and the combined normalized expression value is calculated.

[0174] Typically, the normalized qRT-PCR thresholds for 'altered' expression are 0.7 and 1.99. Normalized protein thresholds (using IHC) are typically 1% and 50% of positive cells. That is, the altered score here refers to tumors where >1% and <50% of tumor cells are positive for normalized gene expression values ​​of at least two of the 11 genes (or proteins). These values ​​can be adjusted based on any new data, but the same theory applies to the terms "normal," "moderate," and "dysregulated" regarding the expression levels of at least two of the 11 genes (or proteins).

[0175] In this instruction manual, the term "adjuvant therapy" should be understood as any treatment administered after primary treatment to increase the chances of long-term survival. The term "neoadjuvant therapy" should be understood as treatment administered before primary treatment to increase the chances of long-term survival. Primary treatment is usually surgery. Adjuvant and neoadjuvant therapies are typically selected from chemotherapy, hormone therapy, targeted therapy, radiation therapy, immunotherapy, or combinations thereof.

[0176] In the instructions, the term "active surveillance" should be understood as close monitoring for cancer. This typically includes a prostate-specific antigen (PSA) blood test approximately every 6 months and a digital rectal exam (DRE) at least once a year. Prostate biopsy and imaging assays may also be performed every 1 to 3 years. Active surveillance may be most suitable if the Gleason score is low (typically 6 or lower), indicating a less aggressive and slower-growing cancer.

[0177] The detection of expression typically involves measuring RNA expression of selected prognostic gene groups in formalin-fixed paraffin-embedded (FFPE) tissue samples from prostate cancer biopsies using quantitative reverse transcriptase polymerase chain reaction (RT-qPCR). Other methods or measurements involve immunohistochemical staining of tumor biopsy or control biopsy tissues using suitable methods, such as immunohistochemical staining. Many other methods for detecting the biomarkers of this invention will be apparent to those skilled in the art. Examples include RNA sequencing, quantitative polymerase chain reaction (qPCR), reverse transcriptase PCR (RT-PCR), quantitative real-time RT-PCR (qRT-PCR), ELISA, Western blotting, and protein assays on polyacrylamide gels.

[0178] In this specification, the term "cancer" should be understood to mean cancer treated with chemotherapy. Examples of such cancers include multiple myeloma, prostate cancer, glioblastoma, lymphoma, fibrosarcoma; myxosarcoma; liposarcoma; chondrosarcoma; osteosarcoma; chordoma; angiosarcoma; endothelial sarcoma; lymphangiosarcoma; lymphangioendothelial sarcoma; synovoma; mesothelioma; Ewing's tumor; leiomyosarcoma; rhabdomyosarcoma; colon cancer; pancreatic cancer; ovarian cancer; squamous cell carcinoma; basal cell carcinoma; adenocarcinoma; sweat gland carcinoma; sebaceous gland carcinoma; papillary carcinoma; papillary adenocarcinoma; cystic adenocarcinoma; medullary carcinoma; bronchial protocarcinoma; renal cell carcinoma; hepatocellular carcinoma; bile duct carcinoma; choriocarcinoma; seminal cell carcinoma. Cystoma, embryonal carcinoma; Wilms' tumor; cervical cancer; uterine cancer; testicular tumor; lung cancer; small cell lung cancer; bladder cancer; epithelial carcinoma; glioma; astrocytoma; medulloblastoma; craniopharyngioma; ependymoma; pineal tumor; hemangioblastoma; acoustic neuroma; oligodendroglioma; meningioma; melanoma; retinoblastoma; head and neck cancer types (in the sinuses, inside and behind the nose, mouth (tongue, gums, palate, posterior oral cavity), throat (nasopharynx, oropharynx, hypopharynx), larynx, lips, salivary glands); bladder cancer types; colorectal cancer; and leukemia.

[0179] In this instruction manual, the term "precancerous lesions" should be understood as referring to abnormal cells that have undergone some changes associated with an increased risk of developing cancer, but are not yet cancer. Examples of precancerous lesions include Barrett's esophagus, Bowen's disease, familial adenomatous polyposis (FAP), lobular carcinoma in situ (LCIS), Lynch syndrome, MEN2, vaginal intraepithelial neoplasia (VAIN), and vulvar intraepithelial neoplasia (VIN). Precancerous lesions are included when diagnosing whether a subject is at risk of developing invasive cancer or at risk of cancer recurrence.

[0180] In this instruction manual, the term "recurrence" should be understood to mean a recurrence of cancer sampled from the patient's body, where the cancer has returned to the sampling area after treatment. For example, if a sample was taken from colon or prostate cancer, it would be a recurrence of colon or prostate cancer in the source tissue. The term should also be understood to mean a recurrence of the primary cancer in a different location than the initially sampled cancer; that is, the cancer has returned to a non-sampling area after treatment, such as a non-localized recurrence.

[0181] In this specification, the term "adverse outcome" should be understood as meaning a very low chance of disease-free survival.

[0182] In the instructions, the term "indolent" should be understood to mean a cancer that typically grows slowly (lazy) and does not cause symptoms or death during an individual's lifetime.

[0183] In this instruction manual, the term "survival rate" should be understood as referring to the likely lifespan of a patient diagnosed with cancer such as colon cancer or prostate cancer. Survival rates are expressed as 5-year, 10-year, 15-year, 20-year, 25-year, 30-year, 35-year, 40-year, 45-year, or 50-year survival rates. Ideally, survival rates are expressed as 5-year or 10-year survival rates.

[0184] In this specification, the term “treatment” should be understood to mean its generally accepted meaning, which encompasses the prevention, suppression, and mitigation, cessation, or reversal of the progression or severity of metastatic, recurrent, or existing cancer phenotypes.

[0185] In this specification, the term "at least one" should be understood to mean and encompass at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, or all genes optionally comprised of the group consisting of: TTC21B, MCM3, MTUS1, FOXM1, CUL4A, GPC1, PCNA, IRF7, BRIP1, ULK1, and SPAG5. Preferably, at least one gene is one, two, or all of TTC21B, MTUS1, MCM3, and FOXM1.

[0186] In this specification, the term "at least two" should be understood to mean and encompass at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, or all genes selected freely from the group consisting of: TTC21B, MCM3, MTUS1, FOXM1, CUL4A, GPC1, PCNA, IRF7, BRIP1, ULK1, and SPAG5. Preferably, the at least two genes are TTC21B and MTUS1, MTUS1 and PCNA, MCM3 and TTC21B, MCM3 and MTUS1, or combinations of these pairs. Preferably, the at least three genes are MTUS1, MCM3, and TTC21B. Preferably, the selected at least four genes are MTUS1, MCM3, TTC21B, and FOXM1.

[0187] Materials and methods

[0188] RNA sequencing

[0189] Total RNA was extracted from proliferating and senescent HMECs using the RNeasy kit (Qiagen). Polyadenylated RNA species were enriched from 5 μg of total RNA, and sequencing libraries were prepared from PolyA+ RNA using the TruSeq Sample Preparation Kit (Illumina). Following the manufacturer's protocol, the libraries were used directly for cluster generation and sequencing analysis using Genome Analyzer II (Illumina). Base calling and mapping to the human genome (hg19) was performed using the BWA sequence alignment tool. In both experiments, the fold change in mRNA was calculated based on the total number of sequence reads mapped to each gene.

[0190] Real-time quantitative PCR

[0191] Total RNA was extracted from cells using the RNeasy kit (Kiagen Biotech) according to the manufacturer's protocol. 1 μg of RNA was used to generate cDNA via reverse transcriptase PCR using the TaqMan reverse transcription kit (Applied Biosystems). Relative mRNA expression levels were determined using SYBR Green I assay chemistry (Applied Biosystems) on an ABI Prism 7500 rapid real-time PCR system. Ribosomal component RPLPO was used as a normalized control gene (SEQ ID NO: XX (forward-CCC) and SEQ ID NO: YY (reverse-CCC). The primer sequence pairs used are shown below (For = forward primer; Rev = reverse primer):

[0192] sequence

[0193] SEQ ID NO:1 (BRIP1; NM_032043):

[0194] aggcgggaattcgtctcgggttgtgtggttgaggggtctggtgggtcgaggaaaggtaacggcggccccagtcctgcacacaaggc

[0195] cggggaagtagcagcacccccaggaagagggaggaggaagggctcgtgccctttcttctcttccagggctccgctttatttgctctca

[0196] gaagtcggtttcctttccttttcttcagtgaatcggagctcagagcgttgcttcggtttccctccagacagttaggaatctgaaataaacag

[0197] gaaagcactatgtcttcaatgtggtctgaatatacaattggtggggtgaagatttactttccttataaagcttacccgtcacagcttgctatg

[0198] atgaattctattctcagaggattaaacagcaagcaacattgtttgttggagagtcccacaggaagtggaaaaagcttagccttactttgttc

[0199] tgctttagcatggcaacaatctcttagtgggaaaccagcagatgagggcgtaagtgaaaaagctgaagtacaattgtcatgttgttgtgc

[0200] atgccattcaaaggattttacaaacaatgacatgaaccaaggaacttcacgtcatttcaactatccaagcacaccaccttctgaaagaaa

[0201] tggcacttcatcaacttgtcaagactcccctgaaaaaaccactctggctgcaaagttatctgctaagaaacaggcatccatatacagaga

[0202] tgaaaatgatgattttcaagtagagaagaaaagaattcgacccttagaaactacacagcagattagaaaacgtcattgctttggaacaga

[0203] agtacacaatttggatgcaaaagttgattcaggaaagactgtaaaactcaactctccactggaaaagataaactccttttcgccacagaa

[0204] accccctggccactgttctaggtgctgttgttctactaaacaaggaaacagtcaagagtcatcgaataccattaagaaggatcatacagg

[0205] gaaatccaagatacccaaaatatattttgggacacgcacacacaagcagattgctcagattactagagagctccggaggacggcatat

[0206] tcaggggttccaatgactattctttccagcagggatcatacttgtgtccatcctgaggtagtcggtaacttcaacagaaatgagaagtgca

[0207] tggaattgctagatgggaaaaacggaaaatcctgctatttttatcatggagttcataaaattagtgatcagcacacattacagactttccaa

[0208] gggatgtgcaaagcctgggatatagaagaacttgtcagcctgggaagaaactaaaggcctgtccatattacacagcccgagaacta

[0209] atacaagatgctgacatcatattttgtccctacaactatctttagatgcacaaataagggaaagtatggatttaaatctgaaagaacaggt

[0210] tgtcattttagatgaagctcataacatcgaggactgtgctcgggaatcagcaagttacagtgtaacagaagttcagcttcggtttgctcgg

[0211] gatgaactatagtatggtcaacaataatataaggaagaaagatcatgaacccctacgagctgtgtgctgtagcctcattaattggtta

[0212] gaagcaaacgctgaatatcttgtagaaagagattatgaatcagcttgtaaaatatggagtggaaatgaaatgctcttaactttacacaaaa

[0213] tgggtatcaccactgctacttttcccattttgcagggacatttttctgctgttcttcaaaaaaggaaaaaatctcaccaatttatggtaaag

[0214] aggaggcaagagaagtacctgttattagtgcatcaactcaaataatgcttaaagactttttatggtacttgactatctttttaggcaaaata

[0215] gcagatttgcagatgattataaaattgcgattcaacagacttactcctggacaaatcagattgatatttcagacaaaaatgggttgttggtt

[0216] ctaccaaaaaataagaaacgttcacgacagaaaactgcagttcatgtgctaaacttttggtgcttaaatccagctgtggccttttcagatat

[0217] taatggcaaagttcagaccattgttttgacatctggtacattatcaccaatgaaatccttttcgtcagaacttggtgttacatttactatccag

[0218] ctggaggctaatcatatcattaaaaattcacaggtttgggttggtaccattgggtcaggccccaagggtcggaatctctgtgctaccttcc

[0219] agaatactgaaacatttgagttccaagatgaagtgggagcacttttgttatctgtgtgccagactgtgagccaaggaattttgtgtttcttg

[0220] ccatcttacaagttattagaaaaattaaaagaacgttggctctctactggtttatggcataatctggagttggtgaagacagtcattgtaga

[0221] accacagggaggagaaaaaacaaattttgatgaattactgcaggtgtactatgacgcaatcaaatacaaaggagagaaagatggagc

[0222] tctcctggtagcagtttgtcgtggtaaagtgagtgagggtctggatttctcagatgacaatgcccgtgctgtcataacaataggaattcctt

[0223] ttccaaatgtgaaagatctacaggttgaactaaaacgacaatacaatgaccaccattcaaaattgagaggtcttctacctggccgtcagt

[0224] ggtatgaaattcaagcatacagggccttaaaccaggcccttggtagatgtattagacacagaaatgattggggagctcttattctagtgg

[0225] atgatcgctttaggaataacccaagtcgctatatatctggactttctaaatgggtacggcagcagattcagcaccattcaacctttgaaag

[0226] tgcactggaatccttggctgaattttccaaaaagcatcaaaaagttcttaatgtatccataaaggacagaaccaatatacaggacaatga

[0227] gtctacacttgaagtgacctctttaaagtacagtacctcaccttatttactggaagcagcaagtcatctatcaccagaaaattttgtggaag

[0228] atgaagcaaagatatgtgtccaggaactacagtgtcctaaaattattaccaaaaattcacctctaccaagtagcattatctccagaaagg

[0229] agaaaaatgatccagtattcctggaagaagcagggaaagcagaaaaaattgtgatttccagatccacaagcccaactttcaacaaaca

[0230] aacaaagagagttagctggtcaagctttaattctttgggacagtattttactggtaaaataccgaaggcaacacctgagctcgggtcatc

[0231] agaatagtgcctctagtcctccccgtttcaaaacagagaagatggaagtaaaaactgttttgcccttcactgataaatgtgaatcctca

[0232] aatctgacagtaaacacatcgtttggatcatgccctcaatcaagaaaccattatttcatcattaaaagattgatgccacccttactagaaaaaa

[0233] tcattctgaaacatccgctctgttctgaagaagccctggatccagacattgaattgtctctagtaagtagaagaataaaagagtccacttca

[0234] aatatagttttgaaaagaagagaagaagaatctatctattttacacctgaactttatgatcctgaagatacagatgaaaaaaaatg

[0235] acctagctgaaactgatagaggaatagattggctaacaattcagattgcattttagctaaagacctttttgaaattaagaactaaaaaga

[0236] gtagattcagccagagagagtgaaaggctgaggattgcatagatcaaagttgaatggaattctgcatattgaagaaagtaaaattgatga

[0237] cattgatggtaatgtaaaaaacaacttggataaatgaactggaactgggaaaaactcatgaaatagaaataagaactttaaaacatctcc

[0238] ttccaaaaaataaaggcatgtttcctggttttaagtaataatacttaactctcaagctaagtaaaaaatatgtcatcatgcttatgttaaactctgt

[0239] tgtaagtaataatttgtaaattgaataagtggcatactttttaaaaaactattttatgttcagaaatgtaaatgttattattcttgagtttttgggtt

[0240] tttttttttgagacagagtcttggtctgttgcccaggctggaatgcagtggtgtgctctgggctcactgcaaccttcacctccaggttcaag

[0241] tgattctcctgcctcagccttctgagtagctgggactacaggtgtgcaccaccatgcccagctagtttttgtatttttagtagagatggggt

[0242] ttcaccatgttggccaggctggtctcgaactcctggcctcgtgatctgcccttctctgcctccctaagttgctgggattacaggtgtgagc

[0243] cacagtgcctggcccattcttgagttttgataaagtaattcatacaaagtactgtcctcaaataagtcttccttagctaaatgcaatttaaaat

[0244] tattcaaagatcctagggcacttctagtttcacgtaaatattcatattaggtggttctcttcatccatttgttttcacactgatacataaaaatta

[0245] acagcagtctaatctagtgacacctcagtcatatttcgctatagattttacctcaaatcagtccaagactttttcagagatcaccatttgtctt

[0246] gaaaggtttatttcgttattaaactgcctacttataagtaattaagagaaattaagaaagtagtatgcatttttaattgaaattgttttacattctt

[0247] tgtataataaacctaaaaccaaacatgtcataaacaaattgacgtaaagatataaaatgccaaatgaagtattccaaattttctattctaatt

[0248] atttagcttcaccatcattgtggaaaaaaatactagatcctgcttagtattatatatttttcctagtggatcagtgagtaataagtaccaaaca

[0249] ctagactagaaggtaatttctacattgtttagaaagggtgaaacaatttatcccctctggtattgttctagcataagctttagttatacaatga

[0250] ttaagatagaaaacttcatatataaatttgataagcaaacccacatttatagctgcagctaaaatatgtttccttagggcacagtaatcctttc

[0251] tgtgaattttgaccttgtttgtgtttttgtgaatgaagctatatgtctaatcaaaaatgattataaagaggctcatctctgacatcattccaaa

[0252] aatacattcattgatctctttttaagaaacatctgttattcactgggcattgggactttttgtgagtaatttgaattgaaattttatgagctatcca

[0253] agaattctgtatggtctattattttcaagtcaaaatttccagtaaggatttactttacatttcatttggataaatgaatcattatataggtatgtctt

[0254] tgcttccattttgagacatttagatttttacagcctgtttctatagcatttgatgttacaactctaagcgtagtttcaaagacatttaaattgacaa

[0255] gttaccagttaaagaatttagaatatattagatcccatctagtattatatattttttctagttgatcattgagcagtaaataccaaatactcgatt

[0256] agaaggtaatttttacattgttttgaaagggtgaaacaatttatctcctctggtattattcttaaaccacagatagggatagtagggtagtga

[0257] aacgaataaatacctggtagaagacaagagacttgggctctacacctggctctgccactgatttgctaagtcatattggcaatcaccaca

[0258] cccttcagggaattagtttcatctgtaaaatgcagcggttagtactataaaatcatacaaatttctttgtgctttgagaatctataaaggaatg

[0259] tctgttgatattctgagtcgattttcatttgctttttgttccagaacggttaaaataaagcatattatttcatttaaaaagtaaagtggctcttactc

[0260] atttacatttagggaacgatggtgacgtggtttggctatgtccccagccagatctcatcttgaattgtagttcccataatcccgacgtgttgt

[0261] gggaaggacccagtgggaggtaattgaatcgtgggggcggttaccctcatgcggttctcgtgatactgagttcttatgagacctgatga

[0262] ttttataaggagcttttccctgcttcactctcattcttctctcctgctgccctgtgaaaagttgccttccgccatgattgtaagtttcttgaggc

[0263] ctcctcagtcatgcagaactgtgagtcaattaaacctctttcctttataaattacccagtcttgggtatgactttattagcagcatgagaaca

[0264] gactgacacagatggtatatgtgcataataacttggaaagctagatatttattttcgcaatgctacaattaaaacattttgaggacttttaaa

[0265] attacctttagggccaggcgcggtggctcacgcctgtaatcccagcactttgggaggccgaggcgggcggatcacgaggtcaggag

[0266] atcgagaccatcctggttaacacggtgaaaccccgtctctactaaaaatacaaaaaattagccaggcgtggtggcgggcgcctgtagt

[0267] cccagctactcaggaggctgaggcaggagaatggcgtgaacccgggaggtggagcttgcagtgagccgagattgcgccactgcac

[0268] tccagcctgggcgacagagcgggactccgtctcaaaaaaaaaaaaaaaaagaaattacctttagaattggtaaactacatgagaaaat

[0269] taagcatattattcatatcttagtgttattacccaacttcatctccaatactttcccctttccctagctagatatgtttttgtttgtttgttttagaca

[0270] gtcttgctctgtcgctcaggctggagtgcaaaggcacgatcttggctcactgcaacctctgcctcgcaggttaaagcgattctcatgtct

[0271] cagcctcctgcgtagctgggattacaggtgtacaccactgcacccagctaatttttttttttttttttttttaagtatcgacagggtttcaccat

[0272] gttggccaggctggtctcaaactcctgacctcaagtgatctgcccacctcggcctcccaaagtgttgggattacaggcgtgagccacc

[0273] atgcctggcctagatattttttattctttccaaaatttaattctccctgaagttaaaaattttcattactgagaatgtacatagagatgtgtcaca

[0274] accctttagtaattccaaaaggtgtttcaaaaattttatacaatgataatcactgttgaaacaggtgtatattctcctcagatcatgactgacc

[0275] aagatgatattccaagaagtaaactactggcctttatagagtaggatgtaggccattttcattactgataacatacttttaaaagaaatgttt

[0276] acagatttaaataagttaaaacatctaaatgcttttaaaagagcacctggcacattgcaagttattcattattaattagtagatgaatcatata

[0277] tctggagtgcacccttgctctatatgaaagcttccctaactatagatatataggtgatagactgacaataaagatttgaggaaagaaaaat

[0278] tatttggctggcttattttttaagcttttgagatgtataaggtagagattgtttatcaaataatttatgtgacacattcaatgcatatgaaataatt

[0279] atctaatactgaatttattcaaggactgaaagtatatagagcacacaggctgcaaaccaggaaaactaccagttagagacagggctttat

[0280] tcttggaataaaggtacaatgtaaagagtagatgtttcatgacttgattaaattatttaaaactgtctagaattgtgttataaaactattactat

[0281] gtttctatgcacttagttattacatgggtttaaatttggcactgtttctaagtttctataaggctttgttgttaagatctttctattcaaaacattaa

[0282] ttttaacaaaaagcttttccccgttatttttctgcaactggacttttttgtattactttcctatattaaaacgcatttaaaata

[0283] SEQ ID NO:2(CUL4;NM_001008895):

[0284] ggttccggcccagccatggcggacgaggccccgcggaagggcagcttctcggcgctcgtgggccgcaccaacggcctcaccaag

[0285] cccgcggccctggccgccgcgcccgccaagccggggggcgcgggcggctccaagaagctggtcatcaagaacttccgagacag

[0286] acctcggctgcccgacaactacacgcaggacacgtggcggaagctgcacgaggcggtgcgggccgtgcagagcagcacctccat

[0287] caggtacaacctcgaggagctctaccaggctgtggaaaatctctgttctcacaaagtctccccaatgctctacaagcaactgcgtcagg

[0288] cctgtgaagaccacgtccaggcacagatccttccgtttagagaagactcactagatagtgttttatttttaaagaagattaacacgtgctg

[0289] gcaggaccactgcagacaaatgatcatgatcagaagcatcttcctgttcttggaccgcacctatgtgctgcagaactccacgctgccct

[0290] ccatctgggatatgggattagaactgtttagaacccatattattagtgataaaatggttcagagtaaaaccattgatggaatcctactgctg

[0291] atcgagcgcgagaggagcggcgaggccgtggaccggagcctgttgcggagcctcctgggcatgctgtctgacctgcaggtgtataa

[0292] agattcatttgaactgaaatttttggaagagactaattgcttatatgctgccgaaggccaaaggttaatgcaggaaagagaggttccaga

[0293] atatcttaaccatgtaagtaaacgcttagaggaagagggagacagagtaatcacttacttggaccacagcacacagaaaccactgatt

[0294] gcttgtgtggagaaacagctattaggagaacatttaacagcaattctgcagaaagggctcgaccacttactggatgagaacagagtgc

[0295] cggacctcgcacagatgtaccagctgttcagccgggtgaggggcgggcagcaggcgctgctgcagcactggagcgagtacatcaa

[0296] gacttttggaacagcgatcgtaatcaatcctgagaaagacaaagacatggtccaagacctgttggacttcaaggacaaggtggaccac

[0297] gtgatcgaggtctgcttccagaagaatgagcggttcgtcaacctgatgaaggagtcctttgagacgttcatcaacaagagacccaaca

[0298] agcctgcagaactgatcgcaaagcatgtggattcaaagttaagagcaggcaacaaagaagccacagacgaggagctggagcggac

[0299] gttggacaagatcatgatcctgttcaggtttatccacggtaaagatgtctttgaagcattttataaaaaagatttggcaaaaagactccttgt

[0300] tgggaaaagtgcctcagtcgatgctgaaaagtctatgttgtcaaagctcaagcatgagtgcggtgcagccttcaccagcaagctggaa

[0301] ggcatgttcaaggacatggagctttcgaaggacatcatggttcatttcaagcagcatatgcagaatcagagtgactcaggccctataga

[0302] cctcacagtgaacatactcacaatgggctactggccaacatacacgcccatggaagtgcacttaaccccagaaatgattaaacttcag

[0303] gaagtatttaaggcattttatcttggaaagcacagtggtcgaaaacttcagtggcaaactactttgggacatgctgttttaaaagcggagtt

[0304] taaagaagggaagaaggaattccaggtgtccctcttccagacactggtgctcctcatgttcaacgagggagatggcttcagctttgagg

[0305] agataaaaatggccacggggatagaggatagtgaattgcgcagaacgctgcagtccctggcctgtggcaaagcacgtgtgctgatta

[0306] aaagtcccaaaggaaaggaagtggaagatggagacaagttcatttttaatggagagttcaagcacaagttgtttagaataaagatcaat

[0307] caaattcagatgaaggaaactgttgaggaacaggttagcaccactgagagagtgtttcaggatagacaatatcagattgatgctgctat

[0308] cgtcagaataatgaagatgagaaagactcttggtcataatcttctagtttctgaattatataatcagctgaaatttccagtaaagcctggag

[0309] atttgaaaaagagaattgaatctctgatagacagagactatatggagagagacaaagacaatccgaatcagtaccactacgtggcctg

[0310] acgcatctgcagacggttccccttcatgaaacactagaatgtaccctcagagcaggaagcacacctgtgccatttctgggactctgatt

[0311] gatccagctgtggacattggaaggcgaaggaagggaggtggctcctgggtcatctttcacaaggctcaagacttcaacctgcagatgt

[0312] atctttttccctccagtttttcctctagttcttttaggcatttaaattgtttctgttactctgtgcaaaaataactttgagattggacaagaagatgtt

[0313] actaaagagaagttcctttaaaaggtcttgttcttgtgtcaaaaagctgcaagtttggtttgttctcgtgtgtgatcatgagtgcacaatgaa

[0314] gaagaccctagatgctgcattttttagctctgaagattccttaggtatccctgaagacagctcgctcagatgatcagcattgagtgaaa

[0315] acaagggcccttcatgggtgaacattagaaagagccagggttcaaagctggcgaatggatgacgcaccctagccactggcccctctc

[0316] tgtttcatgtatttccaaaagttgtaaactttgatggctgatttttcgtaagtcaggtttctaagtgagctccctgaggtgccaaggccatggt

[0317] gtccgccctgctgcgtctgttcgtcagctgagttccttgtgaatctctgttttagggtttggggctagtgtgtttgtgtttccattctaagattg

[0318] agtctggcagtccctgtttttttgcattggggtaactgctctttgattttttttaattgcagtatttgtgtgattgcaataataaagtttggtttggt

[0319] ttttacagtcatgcgcagggacgatccttgttctctgctgtaaactgtaaaaagtttatggagacttaaagtcttgatgttgtgaagcagag

[0320] gttattttgtggaaagattaaaaggattttgttggtacctggttttgtgtgtgtgtgtatatatacatgaggttgaacagtgaaaggaaagttcagt

[0321] agtgatgttagaagggtaactatgacaaagatacttttgagataacatttaaaagtactttatattttacataatagcatgtttcattttgattaa

[0322] aagctaccaaaggaattttgatcatggcataagtgtttaaagcaatattttctggaatataccaagtttatataatttgattttgtgctaaattat

[0323] taagagtctctttttgaaacatgcgggtttgaaatatgacaccttgtgggtttccatattaaaatcctcactctttaattgtcatttctatctttga

[0324] aaattttcatttatgagttccatgatatgtggtctaagaaagaccaaacagatttctattttttttttcttataagttcgttgtgtctagagattgtt

[0325] aatattgtaatttaatgtagacttactttgaataaaattagtttaattggccttaaaattacattaataaaactttgtgatatgcaaatgacacat

[0326] tcttcataactctttagcagctttgactactagtgcactagacagaagtcagagcagggagcacctgtcccctctcaagtgattattctttc

[0327] ctggccacctctaccacaaaataccagtggggccaggtagtgacaagtgggtctttttttacttggacgattcgttgtgctgtgcctttgttt

[0328] catttgagcacagtggctcacgcctgtagtcccagctatgggagggcaaggtgggaggatcgtttgagcctgagttccagaccatcct

[0329] gagcaccatagggagatcctgtttccattttagtttatttatttttaattgaggtggagtcttgctctgttgcccaagctggagtgcagtggtg

[0330] caatctcagctcactgcaccctccgcctcctgggttcaagcagttctcccacctcagtctcccaagtagctgggattacaggcacccgc

[0331] catcatgcccagctaatatttgtatttttgtagagacggggtttcactatgttggccagactggtcttgaactcctaacctcaggtgatctgc

[0332] ccgccttggcctcccaaagtgctgggattacaggcgtgagccaccatgcccggcctgcctgtttctgttttataaaagaaaggaaaatct

[0333] atttatatctattctcttatttagcaatatttaatgttttattgttaaaggtcagtgtggtaaaacaaatagtatagaagaaatcattgaaaaaga

[0334] aacagcacttagacagaaatgctgtaaaagtggaaaggagaaggtgaaattacttgcaggttagatggttgcatacccaggtcatctaa

[0335] ggagcaaatgaaaatctgtgactgaagaaagaattcaccaagttaaccagacacaaaattaacaaaaacatcagtagctttcctataaat

[0336] ataggctataatggagggtcatttttattttatttagagacagggtcttaactgtcacctggactggagtgcagtggcatgatcatgtttca

[0337] cggcagcctctaccttctgggctcaagcgatcctctcttctcagccacctgagtgctgggcctacaggtgtacatcaccacgcccaga

[0338] taatttttattgtattttttatagaggcagtgtcttgctatgttgccaaggctgttcttgtactcctggcctcaagcgaacctcctacctcagcc

[0339] tccccaaatgctgggattacaggcatgagacactgcacctgaccagaagtcattttttcaatagcaacccccaaaaaagataaaatacc

[0340] tagtaacaaatagggaaacacctatgtgaaaggtattataacattactaaggaacacaggccctgaataagtgaagttctgttgatctc

[0341] acattttgctatacagtatcttaagatgtaaaaccttcctttaaaacacaagcggattctaaagttaaaacagaaaagcaaatatgccaga

[0342] ctagtcaaattgttttaaattctgctgtttttagaaacattgtaattgaaacagcatgatgctggccctttaaaaagattcctggaacaagaaa

[0343] gtccacaaataagccaaataacacaggaactcggcgtggcacaaaaggggccgtgtgtgatcaggaactggaactttgcaacaactacc

[0344] catgctcagaagtttgggccaccttgaaaagagaaaagttggattagactcctttttatactaagggaagttccaagtggtcaagtattta

[0345] cgtgtaaaaaaatacatatatatgaaaccataatcataagatactttttttgtaaattgactattattgtataaatttatagagtataaagtgatc

[0346] ttataaattatgcatacattgtggactagtcaagctagttacatccatcacctcaaatacttaacatttttgtagtgagaacatctgaaatttac

[0347] tcttagcaattttgaaatgtacaatactcaatttttgactgtattcaccatgctgtacaatagaactcagaaaaagaaaaatatagtcctcctt

[0348] tctaaga

[0349] SEQ ID NO:3(GPC1;NM_002081):

[0350] gccgagccgggactgcgctagcccgccgcgctctgggctgcccgagcgagcgttcggacctcgcaccccgcgcgccccgcgccg

[0351] ccgccgccgccggcttttgttgtctccgcctcctcggccgccgccgcctctggaccgcgagccgcgcgcgccgggaccttggctctg

[0352] cccttcgcgggcgggaactgcgcaggacccggccaggatccgagagaggcgcgggcgggtggccgggggcgccgccggcccc

[0353] gccatggagctccgggcccgaggctggtggctgctatgtgcggccgcagcgctggtcgcctgcgcccgcggggacccggccagc

[0354] aagagccggagctgcggcgaggtccgccagatctacggagccaagggcttcagcctgagcgacgtgccccaggcggagatctcg

[0355] ggtgagcacctgcggatctgtccccagggctacacctgctgcaccagcgagatggaggagaacctggccaaccgcagccatgccg

[0356] agctggagaccgcgctccgggacagcagccgcgtcctgcaggccatgcttgccacccagctgcgcagcttcgatgaccacttccag

[0357] cacctgctgaacgactcggagcggacgctgcaggccaccttccccggcgccttcggagagctgtacacgcagaacgcgagggcct

[0358] tccgggacctgtactcagagctgcgcctgtactaccgcggtgccaacctgcacctggaggagacgctggccgagttctgggcccgc

[0359] ctgctcgagcgcctcttcaagcagctgcacccccagctgctgctgcctgatgactacctggactgcctgggcaagcaggccgaggcg

[0360] ctgcggcccttcggggaggccccgagagagctgcgcctgcgggccacccgtgccttcgtggctgctcgctcctttgtgcagggcctg

[0361] ggcgtggccagcgacgtggtccggaaagtggctcaggtccccctgggcccggagtgctcgagagctgtcatgaagctggtctactg

[0362] tgctcactgcctgggagtccccggcgccaggccctgccctgactattgccgaaatgtgctcaagggctgccttgccaaccaggccga

[0363] cctggacgccgagtggaggaacctcctggactccatggtgctcatcaccgacaagttctggggtacatcgggtgtggagagtgtcatc

[0364] ggcagcgtgcacacgtggctggcggaggccatcaacgccctccaggacaacagggacacgctcacggccaaggtcatccagggc

[0365] tgcgggaaccccaaggtcaacccccagggccccgggcctgaggagaagcggcgccggggcaagctggccccgcgggagaggc

[0366] caccttcaggcacgctggagaagctggtctccgaagccaaggcccagctccgcgacgtccaggacttctggatcagcctcccaggg

[0367] acactgtgcagtgagaagatggccctgagcactgccagtgatgaccgctgctggaacgggatggccagaggccggtacctccccg

[0368] aggtcatgggtgacggcctggccaaccagatcaacaaccccgaggtggaggtggacatcaccaagccggacatgaccatccggca

[0369] gcagatcatgcagctgaagatcatgaccaaccggctgcgcagcgcctacaacggcaacgacgtggacttccaggacgccagtgac

[0370] gacggcagcggctcgggcagcggtgatggctgtctggatgacctctgcagccggaaggtcagcaggaagagctccagctcccgga

[0371] cgcccttgacccatgccctcccaggcctgtcagagcaggaaggacagaagacctcggctgccagctgcccccagcccccgaccttc

[0372] ctcctgcccctcctcctcttcctggcccttacagtagccaggccccggtggcggtaactgccccaaggccccagggacagaggccaa

[0373] ggactgactttgccaaaaatacaacacagacgatatttaattcacctcagcctggagaggcctggggtgggacagggagggccggc

[0374] ggctctgagcaggggcaggcgcagaggtcccagccccaggcctggcctcgcctgcctttctgccttttaattttgtatgaggtcctcag

[0375] gtcagctgggagccagtgtgcccaaaagccatgtatttcagggacctcaggggcacctccggctgcctagccctccccccagctccc

[0376] tgcaccgccgcagaagcagcccctcgaggcctacagaggaggcctcaaagcaacccgctggagcccacagcgagcctgtgccttc

[0377] ctccccgcctcctcccactgggactcccagcagagcccaccagccagccctggcccaccccccagcctccagagaagccccgcac

[0378] gggctgtctgggtgtccgccatccagggtctggcagagcctctgagatgatgcatgatgccctcccctcagcgcaggctgcagagcc

[0379] cggccccacctccctgcgcccttgaggggccccagcgtctgcagggtgacgcctgagacagcaccactgctgaggagtctgagga

[0380] ctgtcctcccacagacctgcagtgaggggccctccatgcgcagatgaggggccactgacccacctgcgcttctgctggaggagggg

[0381] aagctgggcccaaaggcccagggaggcagcgtgggctctgccaatgtgggctgcccctcgcacacagggctcacagggcaggcc

[0382] ttgctggggtccagggctgttggaggaccccgagggctgaggagcagccaggacccgcctgctcccatcctcacccagatcagga

[0383] accagggcctccctgttcacggtgacacaggtcagggctcagagtgaccctcagctgtcacctgctcacagggatgctggtggctgg

[0384] tgagaccccgcactgcagacgggaatgcctaggtcccttcccgacccagccagctgcagggcacggggacctggatagttaaggg

[0385] cttttccaaacatgcatccatttactgacacttcctgtccttgttcatggagagctgttcgctcctcccagatggcttcggagggccgcag

[0386] ggcccaccttggaccctggtgacctcctgtcactcactgaggccatcagggccctgccccaggcctggacgggccctccttccctcct

[0387] gtgccccagctgccaggcggccctggggaggggtggtgtggtgttgggaaggggtcctgcagggggaggaggacttggagggtc

[0388] tgggggcagctgtcctgaaccgactgaccctgaggaggccgcttagtgctgctttgcttttcatcaccgtcccgcacagtggacggag

[0389] gtccccggttgctggtcaggtccccatggcttgttctctggaacctgactttagatgttttgggatcaggagcccccaacacaggcaagt

[0390] ccaccccataataaccctgccagtgccagggtgggctggggactctggcacagtgatgccgggcgccaggacagcagcactcccg

[0391] ctgcacacagacggcctaggggtggcgctcagaccccaccctacgctcatctctggaaggggcagccctgagtggtcactggtcag

[0392] ggcagtggccaagcctgctgtgtccttcctccacaaggtccccccaccgctcagtgtcagcgggtgacgtgtgttcttttgagtccttgt

[0393] atgaataaaaggctggaaaccta

[0394] SEQ ID NO:4(IRF7;NM_001572):

[0395] gagacgaaacttcccgtcccggcggctctggcacccagggtccggcctgcgccttcccgccaggcctggacactggttcaacacct

[0396] gtgacttcatgtgtgcgcgccggccacacctgcagtcacacctgtagccccctctgccaagagatccataccgaggcagcgtcggtg

[0397] gctacaagccctcagtccacacctgtggacacctgtgacacctggccacacgacctgtggccgcggcctggcgtctgctgcgacag

[0398] gagcccttacctcccctgttataacacctgaccgccacctaactgcccctgcagaaggagcaatggccttggctcctgagagggcagc

[0399] cccacgcgtgctgttcggagagtggctccttggagagatcagcagcggctgctatgaggggctgcagtggctggacgaggcccgca

[0400] cctgtttccgcgtgccctggaagcacttcgcgcgcaaggacctgagcgaggccgacgcgcgcatcttcaaggcctgggctgtggcc

[0401] cgcggcaggtggccgcctagcagcaggggaggtggcccgccccccgaggctgagactgcggagcgcgccggctggaaaacca

[0402] acttccgctgcgcactgcgcagcacgcgtcgcttcgtgatgctgcgggataactcgggggacccggccgacccgcacaaggtgtac

[0403] gcgctcagccgggagctgtgctggcgagaaggcccaggcacggaccagactgaggcagaggcccccgcagctgtcccaccacc

[0404] acagggtgggcccccagggccattcctggcacacacacatgctggactccaagccccaggccccctccctgccccagctggtgaca

[0405] agggggacctcctgctccaggcagtgcaacagagctgcctggcagaccatctgctgacagcgtcatggggggcagatccagtccca

[0406] accaaggctcctggagagggacaagaagggcttcccctgactggggcctgtgctggaggcccagggctccctgctggggagctgt

[0407] acgggtgggcagtagagacgacccccagccccgggccccagcccgcggcactaacgacaggcgaggccgcggccccagagtc

[0408] cccgcaccaggcagagccgtacctgtcaccctccccaagcgcctgcaccgcggtgcaagagcccagcccaggggcgctggacgt

[0409] gaccatcatgtacaagggccgcacggtgctgcagaaggtggtgggacacccgagctgcacgttcctatacggccccccagaccca

[0410] gctgtccgggccacagacccccagcaggtagcattccccagccctgccgagctcccggaccagaagcagctgcgctacacggagg

[0411] aactgctgcggcacgtggcccctgggttgcacctggagcttcgggggccacagctgtgggcccggcgcatgggcaagtgcaaggt

[0412] gtactgggaggtgggcggacccccaggctccgccagcccctccaccccagcctgcctgctgcctcggaactgtgacacccccatctt

[0413] cgacttcagagtcttcttccaagagctggtggaattccgggcacggcagcgccgtggctccccacgctataccatctacctgggcttcg

[0414] ggcaggacctgtcagctgggaggcccaaggagaagagcctggtcctggtgaagctggaaccctggctgtgccgagtgcacctaga

[0415] gggcacgcagcgtgagggtgtgtcttccctggatagcagcagcctcagcctctgcctgtccagcgccaacagcctctatgacgacat

[0416] cgagtgcttccttatggagctggagcagcccgcctagaacccagtctaatgagaactccagaaagctggagcagcccacctagagct

[0417] ggccgcggccgcccagtctaataaaaagaactccagaaca

[0418] SEQ ID NO:5(MCM3;NM_002388):

[0419] Gcgccggggtggagtcatcctgggaacctccacgcgactttggtggaggtagttctttggcagcgggcatggcgggtaccgtggtg

[0420] ctggacgatgtggagctgcgggaggctcagagagattacctggacttcctggacgacgaggaagaccagggaatttatcagagcaa

[0421] agttcgggagctgatcagtgacaaccaataccggctgattgtcaatgtgaatgacctgcgcaggaaaaacgagaagagggctaaccg

[0422] gcttctgaacaatgcctttgaggagctggttgccttccagcgggccttaaaggattttgtggcctccattgatgctacctatgccaagcag

[0423] tatgaggagttctacgtaggactggaaggcagctttggctccaagcacgtctccccgcggactcttacctcctgcttcctcagctgtgtg

[0424] gtctgtgtggagggcattgtcactaaatgttctctagttcgtcccaaagtcgtccgcagtgtccactactgtcctgctactaagaagacca

[0425] tagagcgacgttattctgatctcaccaccctggtggcctttccctccagctctgtctatcctaccaaggatgaggagaacaatccccttga

[0426] gacagaatatggcctttctgtctacaaggatcaccagaccatcaccatccaggagatgccggagaaggccccagccggccagctcc

[0427] cccgctctgtggacgtcattctggatgatgacttggtggataaagcgaagcctggtgaccgggttcaggtggtgggaacctaccgttg

[0428] ccttcctggaaagaagggaggctacacctctgggaccttcaggactgtcctgattgcctgtaatgttaagcagatgagcaaggatgctc

[0429] agccctctttctctgctgaggatatagccaagatcaagaagttcagtaaaacccgatccaaggatatctttgaccagctggccaagtcat

[0430] tggccccaagtatccatgggcatgactatgtcaagaaagcaatcctctgcttgctcttgggaggggtggaacgagacctagaaaatgg

[0431] cagccacatccgtggggacatcaatattcttctaataggagacccatccgttgccaagtctcagcttctgcggtatgtgctttgcactgca

[0432] ccccgagctatccccaccactggccggggctcctctggagtgggtctgacggctgctgtcaccacagaccaggaaacaggagagc

[0433] gccgtctggaagcaggggccatggtcctggctgaccgaggcgtggtttgcattgatgaatttgacaaaatgtctgacatggatcgcac

[0434] agccatccatgaagtgatggagcagggtcgagtgaccattgccaaggctggcatccatgctcggctgaatgcccgctgcagtgttttg

[0435] gcagctgccaaccctgtctacggcaggtatgaccagtataagactccaatggagaacattgggctacaggactcactgctgtcacgat

[0436] ttgacttgctcttcatcatgctggatcagatggatcctgagcaggatcgggagatctcagaccatgtccttcggatgcaccgttacagag

[0437] cacctggggagcaggatggcgatgctatgcccttgggtagtgctgtggatatcctggccacagatgatcccaactttagccaggaaga

[0438] tcagcaggacacccagatttatgagaagcatgacaaccttctacatgggaccaagaagaaaaaggagaagatggtgagtgcagcatt

[0439] catgaagaagtacatccatgtggccaaaatcatcaagcctgtcctgacacaggagtcggccacctacattgcagaagagtattcacgc

[0440] ctgcgcagccaggatagcatgagctcagacaccgccaggacatctccagttacagcccgaacactggaaactctgattcgactggcc

[0441] acagcccatgcgaaggcccgcatgagcaagactgtggacctgcaggatgcagaggaagctgtggagttggtccagtatgcttacttt

[0442] aagaaggttctggagaaggagaagaaacgtaagaagcgaagtgaggatgaatcagagacagaagatgaagaggagaaaagccaa

[0443] gaggaccaggagcagaagaggaagagaaggaagactcgccagccagatgccaaagatggggattcatacgacccctatgacttca

[0444] gtgacacagaggaggaaatgcctcaagtacacactccaaagacggcagactcacaggagaccaaggaatcccagaaagtggagtt

[0445] gagtgaatccaggttgaaggcattcaaggtggccctcttggatgtgttccgggaagctcatgcgcagtcaatcggcatgaatcgcctca

[0446] cagaatccatcaaccgggacagcgaagagcccttctcttcagttgagatccaggctgctctgagcaagatgcaggatgacaatcaggt

[0447] catggtgtctgagggcatcatcttcctcatctgaggaggcctcgtctctgaacttgggttgtgccgagagagtttgttctgtgtttcccacc

[0448] ctctccctgacccaagtctttgcctctactcccttaacagtgttgaattcaactgaaggcgaggaatgttggtgatgaagctgagttcagg

[0449] actcggtggaccctttgggaatgggtcatgaaagctgccatggggtgaggaaagaggagacagtgggagaggacaatgactattgc

[0450] atcttcattgcaaaagcactggctcatccgccctacttcccatcccacacaaacccaattgtaaataacatatgacttctgagtacttttgg

[0451] gggcacaactgttttctgtttgctgtttttttgttttgttttttttctccagagcactttggtctagactaggctttgggtggttccaattggtgga

[0452] gagaagctctgaggcacgtcatgcaggtcaagaaagctttctttgcagtagcaccagttaaggtgaatatgtattgtatcacaaaacaaa

[0453] cccaatatccagatgaatatccgagatgttgaataaacttagccatttcgtacacatgg

[0454] SEQ ID NO:6(MTUS1;NM_001001924):

[0455] aaagggggcggcagcgccggcggagcggaggcgggtctcacgtgggccagcgcagagcctgcggaagggacggatgcggatc

[0456] tcgtcgctgtcaccttgaaagtgaccgaggggcttgactgtggactccttacgccgcccacccgggcccggcggtcccagccttctcg

[0457] cagggccccttctcagcagaagcaagcggggccgagaaagcgggtggaatagggttgctgcaggtcccaaagacccctcgtggcg

[0458] cctcgctactttctgcagcttgtttgcactttttcacgctctagaaaaatctcatcttaattaagggaacaacaaatcatttaatcttcagagc

[0459] atcttagactgaaaacctttcaactgtgctgaaaaacctagaagacagaccattttgcccaccctctcatttaaaaggaattgaagaaga

[0460] aataaaatggcagaggtttaaggttactattcaggatgactgatgataattcagatgataaaatagaagatgaattgcaaaccttctttacc

[0461] agtgataaagatggaaatacacatgcatacaacccgaaatcaccacctacacaaaactcttcagccagcagtgtgaactggaattctgc

[0462] caacccagatgacatggtggttgattatgaaactgaccctgctgtagttactggtgaaaatatttctttaagccttcagggtgttgaagtatt

[0463] tggtcatgaaaagtcttctagtgatttcattagtaagcaggtgttagatatgcataaagattctatttgtcagtgtcctgcacttgtaggtact

[0464] gagaagcccaaatatctgcaacacagttgtcattccctagaagcagttgagggccagagtgttgagccatctttgccttttgtgtggaag

[0465] cctaatgacaatttgaactgtgcaggctactgtgatgccttggagctaaaccaaacatttgacatgacagtggataaagttaactgcacct

[0466] ttatatcacatcatgccatcggaaagagtcagtccttccatactgctggaagcctgccaccaactggtaggagaagtggaagtacatctt

[0467] ctttatcctattccacttggacatcttcccattctgataagacgcatgcaagagaaactacttatgatagagaaagctttgaaaaccctcaa

[0468] gtcacaccatcagaagcccaagacatgacttacacagcattttctgatgtggtgatgcaaagtgaggtttttgtttcagatattggaaatca

[0469] gtgtgcatgttcttcaggaaaggtcaccagtgagtacacagatggatcacaacaaagactagttggagaaaaggagacacaagcact

[0470] aacaccagtttctgatggcatggaagtccccaatgattctgcattacaagagttcttttgtttatcccatgatgaatccaatagcgaaccac

[0471] attcacagagctcatacaggcacaaggaaatgggccaaaatctgagagagacagtgtcctattgtcttattgatgatgaatgccctttaa

[0472] tggtgccagcttttgataagagcgaagctcaagtgctgaacccagagcataaagtcactgagactgaagacacacaaatggtctccaa

[0473] aggaaaggatttgggaacccaaaatcatacctcagaattgattctaagtagcccgccaggacaaaaggtgggctcgtcatttggactg

[0474] acttgggatgcaaatgatatggtcattagcacagacaaaacgatgtgcatgtcaacaccagtcctagaacccacaaaagtaaccttttct

[0475] gtttcaccgattgaagcgacggagaaatgtaagaaagtggagaagggtaatcgagggcttaaaaacataccagactcgaaggaggc

[0476] acctgtgaacctgtgtaaacccagtttaggaaaatcaacaatcaaaacgaataccccaataggctgcaaagttagaaaaactgaaatta

[0477] taagttacccaagaccaaacttcaagaatgtcaaagcaaaagttatgtctagagcagtgttgcagcccaaagatgctgctttatcaaagg

[0478] tcacgcccagacctcagcagaccagtgcctcatcaccctcatcagtgaattcaagacaacaaacagtcttgagcagaacaccgagatc

[0479] tgacttgaatgcagacaaaaaagcagaaattctaattaacaagacacataagcagcagtttaataaactcattactagccaggctgtgca

[0480] tgttacaactcattctaaaaatgcttcacacagggttccaagaacaacatctgccgtgaaatcgaatcaggaagatgttgacaaagcca

[0481] gttcttctaactcagcatgcgagaccgggtccgtttctgcgttgtttcagaagatcaaaggcatactccctgttaaaatggaaagtgcaga

[0482] atgtttggaaatgacctatgttcccaacattgataggattagccctgaaaagaagggtgaaaaagaaaatgggacatctatggaaaaac

[0483] aagagctgaaacaagagattatgaatgagacttttgaatatggttctctgtttttgggctctgcttcaaaaacaacgaccacctcaggtag

[0484] gaatatatccaagcctgactcctgcggtttgaggcaaatagctgctccaaaagccaaagtggggccccctgtttcctgtttgaggcgga

[0485] acagtgacaatagaaatcccagtgctgatcgagccgtatctcctcagaggatcaggcgtgtgtccagttctggaaagcctacatccttg

[0486] aaaactgcacagtcgtcatgggtgaatttgcctagaccacttcctaaatccaaagcatctttgaaaagtcctgcgctgcggaggacagg

[0487] aagcaccccctcaatagccagcacccacagtgagctgagcacttacagcaacaattctggtaatgccgctgtcatcaaatatgaggag

[0488] aaacctccaaaaccagcatttcagaatggttcctcaggatccttttatttgaagcctttggtatccagggctcatgttcacttgatgaaaact

[0489] cctccaaaaggtccttcgagaaaaaatttatttacagctcttaatgcagttgaaaagagcaggcaaaagaatcctcgaagcttatgtatcc

[0490] agccacagacagctcccgatgcgctgccccctgagaaaacacttgaattgacgcaatataaaacaaaatgtgaaaaccaaagtggatt

[0491] tatcctgcagctcaagcagcttcttgcctgtggtaataccaagtttgaggcattgacagttgtgattcagcacctgctgtctgagcgggag

[0492] gaagcactgaaacaacacaaaaccctatctcaagaacttgttaacctccggggagagctagtcactgcttcaaccacctgtgagaaatt

[0493] agaaaaagccaggaatgagttacaaacagtgtatgaagcattcgtccagcagcaccaggctgaaaaaacagaacgagagaatcggc

[0494] ttaaagagttttacaccagggagtatgaaaagcttcgggacacttacattgaagaagcagagaagtacaaaatgcaattgcaagagca

[0495] gtttgacaacttaaatgctgcgcatgaaacctctaagttggaaattgaagctagccactcagagaaacttgaattgctaaagaaggccta

[0496] tgaagcctccctttcagaaattaagaaaggccatgaaatagaaaagaaatcgcttgaagatttactttctgagaagcaggaatcgctaga

[0497] gaagcaaatcaatgatctgaagagtgaaaatgatgctttaaatgaaaaattgaaatcagaagaacaaaaaagaagagcaagagaaaa

[0498] agcaaatttgaaaaatcctcagatcatgtatctagaacaggagttagaaagcctgaaagctgtgttagagatcaagaatgagaaactgc

[0499] atcaacaggacatcaagttaatgaaaatggagaaactggtggacaacaacacagcattggttgacaaattgaagcgtttccagcagga

[0500] gaatgaagaattgaaagctcggatggacaagcacatggcaatctcaaggcagctttccacggagcaggctgttctgcaagagtcgct

[0501] ggagaaggagtcgaaagtcaacaagcgactctctatggaaaacgaggagcttctgtggaaactgcacaatggggacctgtgtagcc

[0502] ccaagagatcccccacatcctccgccatccctttgcagtcaccaaggaattcgggctccttccctagccccagcatttcacccagatga

[0503] cacctccccaaagtccacagactctctgaaagcattttgatgcaggtctgcaggactgaccccaaggaggaacgtgggcacaagagg

[0504] tatatcagcacacgtgtgatcaccgtagggtaactggagcgtcaccaccggcggaatcgcagcttctgagactggaactctggagga

[0505] agacttttgcctccgtccaaaagattcctccaaaaaaagatttaaaaaaagatttcggcatcgacacggacgttgttgcacaaagcactt

[0506] aaagaacgagagcatcttgttcattgcctttttcacctaagcatagggggaaaaactctcagggccctattaagatttataacctttgtaat

[0507] gttcttcaccacagacaccttcttgtgagttttcagtctgactgtgggggtggggggtgtgaatgaaatggatgtcacagagtgtcatgtg

[0508] tctgatgcagcctcctctgctgtgtattaaatgtcaaaatctgaatatatctggatatgtactaatcaaataataatcaatcaatcagcatata

[0509] catttcagccaaagccatagaagaaaaagcaatagttgcttgaattatgatcatctaccaccaactctgctcagccctgtaacagggtag

[0510] ggagagggtataacaggaagagctttgacttgtccctgtctatacattctctgtatcttttgggggtaacttcttggcagtttttcagtgttca

[0511] gccatgtcagttgaaactagatttttctgtagattttttacttacccatgtgagcctaacactatcctgtaattcattttctcaggctatgtgtaa

[0512] atgtagaaccctaatttttctataaaaaaacaaactaactaactaactgtgtaaagaaagaaaaagggaagtaccaatgggtttttccacc

[0513] ttatttttacctttgatctacccttgcagatttaacctgtcttcttccctcccattattctcattttccttttacctttctccaccatccagagccaca

[0514] aaagcaaaccttctacctcctacctacttttctctgggacaaggataaaggaatatgattttccagagccccagagccagctcatcttcca

[0515] ggtgctgaaaccactttccaaataaactaaagcctggatttgatattacaaattttgggaaatcttagaataaagaacgagaacaaggaa

[0516] gtcattggctagtataattaagaaaggtaggattcagtgcttaccgatgatgcagtacttgatagaagaaaacagtctgggaggatagc

[0517] gctcatttttcagttaccctttaaggagtccctttgtctttgggaaagtagcagaatggtccgcttctttcccatgagtggaaaatgtggctt

[0518] gtccaactctcctccaggttgcatttcagtttctttccaaaacttattacctcccctaatcctgagactttggaaaaggtggaaggaagaac

[0519] tgttgctttatctccccctccctgcatgtgtcaacattgtgatgtcagtatttactaatctacattcagtggctgtacaaataacagctgtagt

[0520] aagaagagattcaggatgctagaggtgaatatttgggtcatttacatgtacactacatagcaagttgatactcatgttgcatgttcttttaaa

[0521] ttagtgattttgtgtcttaagtctttaacttccaatacttcatcatgtatgtaaccttccatgtttgcttctgataaatggaaatgtaggttcactg

[0522] ccacttcatgagatatctctgctcacgcttccaagttgttctcaatgacattagccaaagttgggtttgccattcatcccctaggcatggtaa

[0523] atcttgtgttgttccctgctgtcctccgtattacgtgaccggcaaataaatctcatagcagttaatataaaacatctttggaggatgggaga

[0524] gaacaggagggaagatgggaaacaaaatagagaattcttaagattttgtttaaaccaaatgtttcatgtagaatgcaaaatgttggcacg

[0525] tcaaaaatatgaatgtgtagacaactgtagttgtgctcagtttgtagtgatgggaagtgtattttactctgatcaaataaataatgctggaat

[0526] actcaa

[0527] SEQ ID NO:7(PCNA;NM_002592):

[0528] ggatggccggagctggcgccctggttctggaggtaaccggttactgagggcgagaagcgccacccggaggctctagcctgacaaa

[0529] tgcttgctgacctgggccagagctcttcccttacgcaagtctcagccggtcgtcgcgacgttcgcccgctcgctctgaggctcctgaag

[0530] ccgaaaccagctagactttcctccttcccgcctgcctgtagcggcgttgttgccactccgccaccatgttcgaggcgcgcctggtccag

[0531] ggctccatcctcaagaaggtgttggaggcactcaaggacctcatcaacgaggcctgctgggatattagctccagcggtgtaaacctgc

[0532] agagcatggactcgtcccacgtctctttggtgcagctcaccctgcggtctgagggcttcgacacctaccgctgcgaccgcaacctggc

[0533] catgggcgtgaacctcaccagtatgtccaaaatactaaaatgcgccggcaatgaagatatcattacactaagggccgaagataacgcg

[0534] gataccttggcgctagtatttgaagcaccaaaccaggagaaagtttcagactatgaaatgaagttgatggatttagatgttgaacaacttg

[0535] gaattccagaacaggagtacagctgtgtagtaaagatgccttctggtgaatttgcacgtatatgccgagatctcagccatattggagatg

[0536] ctgttgtaatttcctgtgcaaaagacggagtgaaattttctgcaagtggagaacttggaaatggaaacattaaattgtcacagacaagtaa

[0537] tgtcgataaagaggaggaagctgttaccatagagatgaatgaaccagttcaactaacttttgcactgaggtacctgaacttctttacaaaa

[0538] gccactccactctcttcaacggtgacactcagtatgtctgcagatgtaccccttgttgtagagtataaaattgcggatatgggacacttaa

[0539] aatactacttggctcccaagatcgaggatgaagaaggatcttaggcattcttaaaattcaagaaaataaaactaagctctttgagaactg

[0540] cttctaagatgccagcatatactgaagtcttttctgtcaccaaatttgtacctctaagtacatatgtagatattgttttctgtaaataacctatttt

[0541] tttctctattctctgcaatttgtttaaagaataaagtccaaagtcagatctggtctagttaacctagaagtatttttgtctcttagaaatacttgt

[0542] gatttttataatacaaaagggtcttgactctaaatgcagttttaagaattgtttttgaatttaaataaagttacttgaatttcaaacatca

[0543] SEQ ID NO:8(SPAG5;NM_006461):

[0544] aggttcaaacacagacggcgggtgaacatggcgtcctcgacttggtctgagacgtgataggcctgccttctggttgaagatgtggcga

[0545] gtgaaaaaactgagcctcagcctgtcgccttcgccccagacgggaaaaccatctatgagaactcctctccgtgaacttaccctgcagc

[0546] ccggtgccctcaccaactctggaaaaagatcccccgcttgctcctcgctgaccccatcactgtgcaagctggggctgcaggaaggca

[0547] gcaacaactcatctccagtggattttgtaaataacaagaggacagacttatcttcagaacatttcagtcattcctcaaagtggctagaaac

[0548] ttgtcagcatgaatcagatgagcagcctctagatccaattccccaaattagctctactcctaaaacgtctgaggaagcagtagacccact

[0549] gggcaattatatggttaaaaccatcgtccttgtaccatctccactggggcagcaacaagacatgatatttgaggcccgtttagataccat

[0550] ggcagagacaaacagcatatctttaaatggacctttgagaacagacgatctggtgagagaggaggtggcaccctgcatgggagaca

[0551] ggttttcagaagttgctgctgtatctgagaaacctatctttcaggaatctccgtcccatctcttagaggagtctccaccaaatccctgttctg

[0552] aacaactacattgctccaaggaaagcctgagcagtagaactgaggctgtgcgtgaggacttagtaccttctgaaagtaacgccttcttg

[0553] ccttcctctgttctctggctttccccttcaactgccttggcagcagatttccgtgtcaatcatgtggacccagaggaggaaattgtagagc

[0554] atggagctatggaggaaagagaaatgaggtttcccacacatcctaaggagtctgaaacagaagatcaagcacttgtctcaagtgtgga

[0555] agatattctgtccacatgcctgacaccaaatctagtagaaatggaatcccaagaagctccaggcccagcagtagaagatgttggtagg

[0556] attcttggctctgatacagagtcttggatgtccccactggcctggctggaaaaaggtgtaaatacctccgtcatgctggaaaatctccgc

[0557] caaagcttatcccttccctcgatgcttcgggatgctgcaattggcactacccctttctctacttgctcggtggggacttggtttactccttca

[0558] gcaccacaggaaaagagtacaaacacatcccagacaggcctggttggcaccaagcacagtacttctgagacagagcagctcctgtg

[0559] tggccggcctccagatctgactgccttgtctcgacatgacttggaagataacctgctgagctctcttgtcattctggaggttctctcccgc

[0560] cagcttcgggactggaagagccagctggctgtccctcacccagaaacccaggacagtagcacacagactgacacatctcacagtgg

[0561] gataactaataaacttcagcatcttaaggagagccatgagatgggacaggccctacagcaggccagaaatgtcatgcaatcatgggtg

[0562] cttatctctaaagagctgatatccttgcttcacctatccctgttgcatttagaagaagataagactactgtgagtcaggagtctcggcgtgc

[0563] agaaacattggtctgttgctgttttgatttgctgaagaaattgagggcaaagctccagagcctcaaagcagaaagggaggaggcaagg

[0564] cacagagaggaaatggctctcagaggcaaggatgcggcagagatagtgttggaggctttctgtgcacacgccagccagcgcatcag

[0565] ccagctggaacaggacctagcatccatgcgggaattcagaggccttctgaaggatgcccagacccaactggtagggcttcatgccaa

[0566] gcaagaagagctggttcagcagacagtgagtcttacttctaccttgcaacaagactggaggtccatgcaactggattatacaacatgga

[0567] cagctttgctgagtcggtcccgacaactcacagagaaactcacagtcaagagccagcaagccctgcaggaacgtgatgtggcaattg

[0568] aggaaaagcaggaggtttctagggtgctggaacaagtctctgcccagttagaggagtgcaaaggccaaacagaacaactggagttg

[0569] gaaaacagtcgtctagcaacagatctccgggctcagttgcagattctggccaacatggacagccagctaaaagagctacagagtcag

[0570] catacccattgtgcccaggacctggctatgaaggatgagttactctgccagcttacccagagcaatgaggagcaggctgctcaatggc

[0571] aaaaggaagagatggcactaaaacacatgcaggcagaactgcagcagcaacaagctgtcctggccaaagaggtgcgggacctgaa

[0572] agagaccttggagtttgcagaccaggagaatcaggttgctcacctggagctgggtcaggttgagtgtcaattgaaaaccacactggaa

[0573] gtgctccgggagcgcagcttgcagtgtgagaacctcaaggacactgtagagaacctaacggctaaactggccagcaccatagcaga

[0574] taaccaggagcaagatctggagaaaacacggcagtactctcaaaagctagggctgctgactgagcaactacagagcctgactctcttt

[0575] ctacagacaaaactaaaggagaagactgaacaagagacccttctgctgagtacagcctgtcctcccacccaggaacaccctctgcct

[0576] aatgacaggaccttcctgggaagcatcttgacagcagtggcagatgaagagccagaatcaactcctgtgcccttgcttggaagtgaca

[0577] agagtgctttcacccgagtagcatcaatggtttcccttcagcccgcagagaccccaggcatggaggagagcctggcagaaatgagta

[0578] ttatgactactgagcttcagagtctttgttccctgctacaagagtctaaagaagaagccatcaggactctgcagcgaaaaatttgtgagct

[0579] gcaagctaggctgcaggcccaggaagaacagcatcaggaagtccagaaggcaaaagaagcagacatagagaagctgaaccagg

[0580] ccttgtgcttgcgctacaagaatgaaaaggagctccaggaagtgatacagcagcagaatgagaagatcctagaacagatagacaaga

[0581] gtggcgagctcataagccttagagaggaggtgacccaccttacccgctcacttcggcgtgcggagacagagaccaaagtgctccag

[0582] gaggccctggcaggccagctggactccaactgccagcctatggccaccaattggatccaggagaaagtgtggctctctcaggaggt

[0583] ggacaaactgagagtgatgttcctggagatgaaaaatgagaaggaaaaactcatgatcaagttccagagccatagaaatatcctagag

[0584] gagaaccttcggcgctctgacaaggagttagaaaaactagatgacattgttcagcatatttataagaccctgctctctattccagaggtg

[0585] gtgaggggatgcaaagaactacagggattgctggaatttctgagctaagaaactgaaagccagaatctgcttcacctctttttacctgca

[0586] atacccccttaccccaataccaagaccaactggcatagagccaactgagataaatgctatttaaataaagtgtatttaatgaa

[0587] SEQ ID NO:9(TTC21B;NM_024753):

[0588] atcatgtattactggttgccagtgaaggaattaagaggtatggaagtgatccagtcttcaggttttatcatgcctatggcacattaatggaa

[0589] ggtaaaactcaagaagctcttcgagaatttgaggctattaaaaataaacaagatgtatcactttgttctctacttgcactgatatatgcccat

[0590] aaaatgagtcctaatccagatagagaagctattctggaatcagatgccagagtgaaggaacaacgtaaaggagctggagagaaagc

[0591] cttataccatgcaggcttatttttatggcacattggtcgccatgataaagcaagggaatatattgacagaatgatcaaaatatcagatggta

[0592] gtaaacagggacacgttttgaaagcatggcttgatattacaagaggaaaagagccttacactaaaaaagcactgaagtattttgaagag

[0593] ggactccaagatgggaatgatacttttgctctgctgggtaaggcacaatgccttgagatgcgccagaattattcaggtgccctggagac

[0594] tgtgaaccagataatcgtgaattttccgagcttccttcctgcttttgttaagaaaatgaaattacaactagccttgcaggattgggaccaga

[0595] cagttgagacagcacaaaggttgctgctccaagatagccaaaatgtggaagcactgagaatgcaggcactctactatgtgtgtagaga

[0596] gggggatatagagaaggcttccaccaagctggaaaacttgggaaatacattggatgccatggaaccacagaatgctcaacttttctata

[0597] acattacactcgccttcagcagaacttgtggacgtagtcaacttattcttcaaaaaattcaaacgttacttgagagagcttttagtttaaacc

[0598] ctcagcaatcagaatttgctacagaacttggataccaaatgattttacaaggaagagttaaagaggcactgaagtggtataagaccgcc

[0599] atgacacttgatgagactagtgtgtctgccctagttggatttatccaatgtcagttgatagaagggcaattacaggatgcagatcagcag

[0600] ctagaatttttaaatgaaatccagcaatccattggaaaatctgcggaattaatctatttacatgcagttcttgccatgaagaaaaataaacg

[0601] acaagaagaagttattaatttgttaaatgatgtcctggacactcacttttcacaattagaaggtttgcctcttggcatacagtattttgaaaag

[0602] ctaaatcctgatttcttgttagaaattgttatggagtatctgagcttctgtccaatgcagcctgcaagtcctgggcaacctctttgtccacttc

[0603] tcaggcgttgcatctcagtcctggagactgtagtaagaactgttccaggtcttctgcaaacagtcttcctaatagcaaagtgaaatatttg

[0604] tcaggtgatattgaagcagctttcaataaccttcagcactgcttagaacacaatccctcttatgctgatgctcatctgctgctagctcaggtt

[0605] tacttgtctcaagaaaaagtcaaattgtgttctcagtctcttgaactttgtctgagctatgattttaaggtgagagactatcctttataccattt

[0606] gataaaagctcagtcacaaaagaaaatgggagaaaatagcagacgcaattaaaacactgcatatggcaatgagtttaccaggaatgaa

[0607] aagaattggagcttccacaaaatcaaaagacagaaaaaactgaagttgatacaagccatcgtttatcgatctttcttgaattgatagacgtt

[0608] caccgcttaaatggagagcagcatgaggcaaccaaagttttacaagatgccatccatgaattttctggaacatctgaagaagtgcgggt

[0609] taccattgctaatgcagaccttgctctagcccaaggatattgaacgggctttaagcatccttcagaatgttacagccgaacagccttat

[0610] tttatagaggccagagaaaaaatggcagatatttatctgaagcacagaaaagataaaatgttatatatcacttgttttagagaaattgctga

[0611] aagaatggctaaccctcggtctttctccttggtgatgcatacatgaatattctagagcctgaagaagccatagtagcatatgagcaa

[0612] gcattaaatcagaacccgaaagatggaacattggcaagcaaaatgggcaaagcacttaatcaaaactcataactactcaatggcaatca

[0613] cttactatgaagctgctctgaaaactggacaaaagaattatctttgctatgacctggctgagctctttataaaattgaaatggtatgacaaa

[0614] gcagaaaaagttcttcagcatgctctggctcatgaacctgtaaatgaactgtcagctctcatggaggatggacgttgtcaagttcttctag

[0615] caaaagtttatagtaaaatggaaaaacttggtgatgcgatcactgcattacaacaggctcgagaattacaagctcgggtactaaaacgt

[0616] gttcagatggaacagccagatgcagttcctgcacagaaacatttagcagctgaaatttgtgcagagattgcaaacattctgttgctcag

[0617] cgagactatgaaaaagcaattaagttttatagagaggctctggttcactgcgaaacagataataagattatgttggaactggcacgattat

[0618] acctggcacaagatgaccctgattcctgcctgcggcagtgtgctctactgcttcagagtgaccaggataacgaagctgctaccatgatg

[0619] atggctgatctcatgttcagaaaacaagactatgaacaagcagtgtttcatttacagcagcttttagaacgtaagccagataattatatgac

[0620] attatctcgtttgattgatctcctaagaagatgtggaaaaactcgaggatgtcccaagatttttctcaatggctgaaacgtaactccagag

[0621] caaaattggaaccaggatttcagtattgtaaaggactgtatctttggtacactggagaaccaaatgatgcccttcgacattttaataaagct

[0622] cggaaagatcgtgactggggccaaaatgccctttataatatgatagagatctgtttgaatccagataatgaaactgttggaggtgaagta

[0623] tttgaaaacctggatggagacctgggtaattcaactgagaagcaagaatctgtgcaactggcagtaagaacagcagaaaaacttcttaa

[0624] ggaactaaaacctcagactgttcagggtcacgtacagcttcgcataatggaaaactattgcttaatggctaccaaacagaaatctaatgt

[0625] tgaacaagcattaaataccttcactgaaatagcagcatctgagaaggagcatatcccagcgctcttgggaatggcaacggcttatatga

[0626] tcttgaaacagactccacgagccagaaaccagctgaagcgtattgcgaaaatgaattggaatgctattgatgctgaagagtttgagaag

[0627] agttggctgctacttgctgatatttacattcaatcagcaaaatatgacatggcagaagacctgttaaaacggtgcctgcgtcataatagat

[0628] cttgctgcaaagcttatgaatatatgggatacattatggaaaaagagcaagcatatacagatgctgccttgaactatgagatggcatgga

[0629] aatatagcaatcggacaaatccggcagtaggatacaaactggcatttaattacttaaaagcaaaaagatatgtggattcaattgacatat

[0630] gtcaccaggttcttgaagcacatccaacttatccaaaaatcagaaaggatatacttgataaggcccgtgcgtctttaagaccttgaaaaata

[0631] attttaacttaggtgttggtttaacaggaaatgaaagaaatctaactttcagttcttcctgttcaaaacaggtttgagctcagtgctttgttatt

[0632] agaagtatacccttttttctccagcagaggttgctgctgtacatcaagagaagtactatgtgaaattggtgtttcctaatggagttgaatga

[0633] gagctggtctatttgactctgttttgattgggtacagatttggtgactctgtggtaaagactataattatttctataaagaattttgttaaaat

[0634] ctaggtaattaaataccctgtatcttttctaaggaatattatttcaggaaatatatttaaaatgcattgttctcttttaaagtgtttttgttatattctt

[0635] ttatttatttatttatttttaaagacagagccttgatctgccgcccaggctggagtgcagtggcacgatcttggctcactgcaacctccgcct

[0636] cctgggtttacgcgattctcctgcttcagcctcccgagtagctgggactaaaggcatgtgccatcacacccggctaattttttgtattttta

[0637] gtagagatgggatttcaccgtgttagccaggaggggcttgatctcttgaccttgtgatccacctgcctcggcctcccaaagtgctgggat

[0638] tacaggcatgagccaccgcacccagcctgtttttgttgtattctgtgttgtttcaaaaattggaataaatctccctgtaaataactcatgaaa

[0639] aagacaatatatattttaaatatatattttaataaaggttttataaatagatgatttatattcctcttggaactcaaaagatgtggtaagttttg

[0640] caaatcagggtaagctccccctgcattctgtaatgttaatgggtggtggtatagtacagaaaatgtaactttcattgaaaaatcttttttaga

[0641] aaagggtagagtgtgtgtttccctttcctttcttgagctcattcttcctgctttcaaggccattgtggtcttaagaaataatcaggtcctgaaa

[0642] cctcaatttaaataggtcccttattagggatatttaaccttagcctagaagacttggtctgcaggtataatttcagacactgtaacatttgatg

[0643] gtaaagggtgaccaaatacttttggtcagcagggagtcacaaagcacaccatgtgaaggcaacggtttacccagctctgtttaacagc

[0644] accctggcatagaagctttcactctcttacactaaatttaacctaatctaaacttaatccttagcattatttttaatttctcatctacatctttttca

[0645] attttccttaaaattgtagtaaacacagtcctcaattttatttgcatttacaataaacagtatttataagtca

[0646] SEQ ID NO:10(ULK1;NM_003565):

[0647] agtgcggctgcgcgggcgtctcaggctctgaggcccgggcgccgcggctcttttgtttctccgttggggccgagcccgggccctagt

[0648] tggagccggagtcggagtcggagtcggatccggattcggattagcagcccgggaagagtgccgtggcacaggcgccggagggag

[0649] cgcgaccctcggaccccgcctggcccgcggggctgggacccggccccggcctgcccgatggggcgcgcggccccggagatgc

[0650] gccctcgcccggccccgcgcccccggccccgcgcccccggcccgcccgccccggcccgcgcctccgcctgagtcccccgcgcc

[0651] ttggcccgccaccccccgccccgcgcccccggcccgcctgcgccatggagcccggccgcggcggcacagagaccgtgggcaag

[0652] ttcgagttctcccgcaaggacctgatcggccacggcgccttcgcggtggtcttcaagggccgccaccgcgagaagcacgatttggag

[0653] gtcgccgtcaagtgcattaacaagaagaacctcgccaagtctcagacgctgctggggaaggaaatcaaaatcctgaaggaactgaaa

[0654] catgaaaacatcgtggccctgtacgacttccaggaaatggctaattctgtctacctggttatggagtactgcaacggtggggacctggc

[0655] cgactacctgcacgccatgcgcacgctgagcgaggacaccatcaggctcttcctgcagcagatcgcgggcgccatgcggcttctgc

[0656] acagcaaaggcatcatccaccgcgacctgaaaccgcagaacatcctgctgtccaaccccgccggccgccgcgccaaccccaacag

[0657] catccgcgtcaagatcgctgacttcggcttcgcgcggtacctccagagcaacatgatggcggccacactctgcggctcccccatgtac

[0658] atggcccccgaggtcatcatgtcccagcactacgacgggaaggcggacctgtggagcatcggcaccatcgtctaccagtgcctgac

[0659] ggggaaggcgcccttccaggccagcagcccccaggacctgcgcctgttctacgagaagaacaagacgttggtccccaccatcccc

[0660] cgggagacctcggccccgctgcggcagctgctcctggccctactgcaacgcaaccacaaggaccgcatggacttcgatgagtttttt

[0661] catcaccctttcctcgatgccagcccctcggtcaggaaatccccacccgtgcctgtgccctcgtacccaagctcggggtccggcagca

[0662] gctccagcagcagctccacctcccacctggcctccccgccgtccctgggcgagatgcagcagctgcagaagaccctggcctccccg

[0663] gctgacaccgctggcttcctgcacagctcccgggactctggtggcagcaaggactcttcctgtgacacagacgacttcgtcatggtcc

[0664] ccgcgcagtttccaggtgacctggtggctgaggcgcccagtgccaaacccccgccagacagcctgatgtgcagtgggagctcactg

[0665] gtggcctctgcgggcttggagagccacggccggaccccatctccatccccaccctgcagcagctcccccagtccctcaggccgggc

[0666] tggcccgttctccagcagcaggtgcggcgcctctgtccccatcccagtccccacgcaggtgcagaactaccagcgcattgagcgaaa

[0667] cctgcagtcacccacccagttccaaacacctcggtcctctgccatccgcaggtcaggcagcaccagccccctgggctttgcaagggc

[0668] cagcccctcgccccctgcccacgctgagcatggaggcgtcctggccaggaagatgtctctgggtggaggccggccctacacgccat

[0669] ctcctcaagttggaaccatccctgagcggccaggctggagcgggacgccctccccacagggagctgagatgcggggtggcaggtc

[0670] ccctcgtccaggctcctctgcacccgagcactctccccgcacttccgggctgggctgccgcctgcacagcgcccccaacctgtctga

[0671] cttgcacgtcgtccgccccaagctgcccaaaccccccacggaccccctgggagctgtgttcagcccaccacaggccagccctcccc

[0672] agccgtcccacggcctgcagtcctgccggaacctgcggggctcacccaagctgcccgacttcctgcagcgaaaccccctgcccccc

[0673] atcctgggctcccccaccaaggctgtgccctcctttgacttcccgaagacccccagctcccagaacctgctggccctcctagcccggc

[0674] agggcgtggtgatgacgccccctcgaaaccggacgctgcccgacctctcggaggtgggacccttccatggtcagccgttgggccct

[0675] ggcctgcggccaggcgaggaccccaagggcccctttggccggtctttcagcaccagccgcctcactgacctgctccttaaggcggc

[0676] gtttgggacacaagccccggacccgggcagcacggagagcctgcaggagaagcccatggagatcgcaccctcagctggctttgga

[0677] gggagcctgcacccaggagcccgtgctgggggcaccagcagcccttccccggtggtcttcaccgtgggctctcccccgagcggga

[0678] gcacgcccccccagggcccccgcaccaggatgttctcagcgggccccactggctctgccagctcttctgcccgccacctggtgcctg

[0679] ggccctgcagcgaggccccagcccctgagctccctgctccaggacacggctgcagctttgccgaccccattactgcgaacctggag

[0680] ggggctgtgaccttcgaggcccccgacctccctgaggagaccctcatggagcaagagcacacggagatcctgcgtggcctgcgctt

[0681] cacgctgctgttcgtgcagcacgtcctggagatcgcagccctgaagggcagcgccagtgaggcggcggggggccctgagtaccag

[0682] ctgcaggagagtgtggtggccgaccagatcagcctgctgagccgagaatggggcttcgcggaacagctggtgctgtacctgaaggt

[0683] ggccgagctactgtcctccggcctgcaaagtgccatcgaccagatccgggccggcaagctctgcctgtcgtccactgtgaagcaggt

[0684] ggtgcgcaggctgaatgagctgtacaaggccagcgtggtgtcctgccagggcctgagcctgcggctgcagcgcttcttcctggacaa

[0685] gcagcggctcctggaccgcattcacagcatcactgccgagaggctcatcttcagccacgctgtgcagatggtgcagtcggctgccct

[0686] ggacgagatgttccagcaccgtgagggctgcgtcccacgctaccacaaggccctgctgctcctggaggggctgcagcacatgctct

[0687] cggaccaggccgacatcgagaacgtcaccaagtgcaagctgtgcattgagcggagactctcggcgctgctgactggcatctgtgcct

[0688] gacctttctggcctggctgggccccccgtcctgccgagccctgcagagtgggctctgtgtgctggctggactcctcgggacaagccc

[0689] atggcgctgatcgctggtgctgagccctgccctgggccccacggacagtcagcctgccggcctccctgcagctcacggggcagaac

[0690] cagcacatctggagccacacagcttggggggtgtctcccatcttttacaggtggggatcacagaatttctgcccctccagctgcctggc

[0691] tcagcaggcgtgggtgccaccaccctctagccccagggcagccccggaggacaggcaagggcctgagaccactgccgactcaaa

[0692] gccaaagcgagctcctgcttagggaaggtcagcaggcactgtgcccaggaagagcctgcggcctcggcgtcccccagtctccagg

[0693] agcctctccctccgagatacccacccagctttgtcaatcacccaagcactttatgcatatagagacagaacctggacctcaccagggac

[0694] tgctgggcagcgattcctggcagtggcctggtgtttgtacatacacatatgcagacacatgccagggccccccaagcccgagcaccg

[0695] gaccacgttgctgcccaggtctggacctcagcgggagaactggctccggggggagtggggccctgcgctagaggcagaggcagtt

[0696] ctttgttcaagcgttcctctggggaccggcagcagaggcaccgtgttctctcagccctggatacgtcttgtaatctttcacactttattccta

[0697] aaacgtgtcttatttttatgcagctcattttttctttaaaggagaaaacttgtaggtgtttaagaattggttttgggagggcgaggactgggc

[0698] caggttagaggcagatggcacaggggcgtgtggcgggcgggtgaggctgctttgcacacctgtgttggtggctgtcccctgccgcc

[0699] cctccctgtggcagcagcaggacaggtgtgtgcccagcaccctccctacctgggcctggaagcagatgaggggaatacttcatgcaa

[0700] agaaaaaagtaacatgtgcaaaagctccccgtccagctttgacagtcagttttgatgtcagctcctcggcagggtaggcctgatgacag

[0701] ccctgtccctccctgcctctgccttgcccaaggccacggagggcatctgcagagaggcctgccttccggattccaggcgggcatgcc

[0702] ctgcaaaccccgcctgggcctcccttggtctgcccagccctcgattagccctgcctgaatcagtagatacttgaacgagtccccagtct

[0703] gcgggaggcagtggtggggccatggacccatgcggggggttccagggtcacacgccacataacagacaaaaatacacacacgtgt

[0704] gtttttctttgcaatacttgaaatattgccactgtgcttggacttagaagaagaaaatccccgtgacttcttcctcatcaccttgatggctttat

[0705] tctcaccttgtggggcatgtttgtatttattgcttcatggccgactggaatcctgagtcctgggaagctggcactgcggggatcttgcccg

[0706] gtgtcctggtcctcttgcttccgtcgcggccgcatgtgcgtgtgtccaagcaggtcctgggcgcctcaactgctgcccctggttgaatgt

[0707] tctcttgatagtgctggaccctttgtctattttaaagcgaattttgtgtgatttcctgccctttgcgttatattgtataataccaacgtaaggaaa

[0708] taaacctttggaattgtt

[0709] SEQ ID NO:11(FOXM1;NM_202002):

[0710] tttcaaacagcggaacaaactgaaagctccggtgccagaccccacccccggccccggcccgggaccccctcccctcccgggatcc

[0711] cccggggttcccaccccgcccgcaccgccggggacccggccggtccggcgcgagcccccgtccggggccctggctcggccccc

[0712] aggttggaggagcccggagcccgccttcggagctacggcctaacggcggcggcgactgcagtctggagggtccacacttgtgattc

[0713] tcaatggagagtgaaaacgcagattcataatgaaaactagcccccgtcggccactgattctcaaaagacggaggctgccccttcctgtt

[0714] caaaatgccccaagtgaaacatcagaggaggaacctaagagatcccctgcccaacaggagtctaatcaagcagaggcctccaagg

[0715] aagtggcagagtccaactcttgcaagtttccagctgggatcaagattattaaccaccccaccatgcccaacacgcaagtagtggccatc

[0716] cccaacaatgctaatattcacagcatcatcacagcactgactgccaagggaaaagagagtggcagtagtgggcccaacaaattcatcc

[0717] tcatcagctgtgggggagccccaactcagcctccaggactccggcctcaaacccaaaccagctatgatgccaaaaggacagaagtg

[0718] accctggagaccttgggaccaaaacctgcagctagggatgtgaatcttcctagaccacctggagccctttgcgagcagaaacgggag

[0719] acctgtgcagatggtgaggcagcaggctgcactatcaacaatagcctatccaacatccagtggcttcgaaagatgagttctgatggact

[0720] gggctcccgcagcatcaagcaagagatggaggaaaaggagaattgtcacctggagcagcgacaggttaaggttgaggagccttcg

[0721] agaccatcagcgtcctggcagaactctgtgtctgagcggccaccctactcttacatggccatgatacaattcgccatcaacagcactga

[0722] gaggaagcgcatgactttgaaagacatctatacgtggattgaggaccactttccctactttaagcacattgccaagccaggctggaaga

[0723] actccatccgccacaacctttccctgcacgacatgtttgtccgggagacgtctgccaatggcaaggtctccttctggaccattcacccca

[0724] gtgccaaccgctacttgacattggaccaggtgtttaagccactggacccagggtctccacaattgcccgagcacttggaatcacagca

[0725] gaaacgaccgaatccagagctccgccggaacatgaccatcaaaaccgaactccccctgggcgcacggcggaagatgaagccactg

[0726] ctaccacgggtcagctcatacctggtacctatccagttcccggtgaaccagtcactggtgttgcagccctcggtgaaggtgccattgcc

[0727] cctggcggcttccctcatgagctcagagcttgcccgccatagcaagcgagtccgcattgcccccaaggtttttggggaacaggtggtg

[0728] tttggttacatgagtaagttctttagtggcgatctgcgagattttggtacacccatcaccagcttgtttaattttatctttctttgtttatcagtgc

[0729] tgctagctgaggaggggatagctcctctttcttctgcaggaccagggaaagaggagaaactcctgtttggagaagggttttctcctttgc

[0730] ttccagttcagactatcaaggaggaagaaatccagcctggggaggaaatgccacacttagcgagacccatcaaagtggagagccctc

[0731] ccttggaagagtggccctccccggccccatctttcaaagaggaatcatctcactcctgggaggattcgtcccaatctcccaccccaaga

[0732] cccaagaagtcctacagtgggcttaggtccccaacccggtgtgtctcggaaatgcttgtgattcaacacagggagaggagggagagg

[0733] agccggtctcggaggaaacagcatctactgcctccctgtgtggatgagccggagctgctcttctcagaggggcccagtacttcccgct

[0734] gggccgcagagctcccgttcccagcagactcctctgaccctgcctcccagctcagctactcccaggaagtgggaggaccttttaaga

[0735] cacccattaaggaaacgctgcccatctcctccaccccgagcaaatctgtcctccccagaacccctgaatcctggaggctcacgccccc

[0736] agccaaagtagggggactggatttcagcccagtacaaacctcccagggtgcctctgaccccttgcctgaccccctggggctgatgga

[0737] tctcagcaccactcccttgcaaagtgctcccccccttgaatcaccgcaaaggctcctcagttcagaacccttagacctcatctccgtccc

[0738] ctttggcaactcttctccctcagatatagacgtccccaagccaggctccccggagccacaggtttctggccttgcagccaatcgttctct

[0739] gacagaaggcctggtcctggacacaatgaatgacagcctcagcaagatcctgctggacatcagctttcctggcctggacgaggaccc

[0740] actgggccctgacaacatcaactggtcccagtttattcctgagctacagtagagccctgcccttgcccctgtgctcaagctgtccaccat

[0741] cccgggcactccaaggctcagtgcaccccaagcctctgagtgaggacagcaggcagggactgttctgctcctcatagctccctgctg

[0742] cctgattatgcaaaagtagcagtcacaccctagccactgctgggaccttgtgttccccaagagtatctgattcctctgctgtccctgccag

[0743] gagctgaagggtgggaacaacaaaggcaatggtgaaaagagattaggaaccccccagcctgtttccattctctgcccagcagtctctt

[0744] accttccctgatctttgcagggtggtccgtgtaaatagtataaattctccaaattatcctctaattataaatgtaagcttatttccttagatcatt

[0745] atccagagactgccagaaggtgggtaggatgacctggggtttcaattgacttctgttccttgcttttagttttgatagaagggaagacctg

[0746] cagtgcacggtttcttccaggctgaggtacctggatcttgggttcttcactgcagggacccagacaagtggatctgcttgccagagtcct

[0747] ttttgcccctccctgccacctccccgtgtttccaagtcagctttcctgcaagaagaaatcctggttaaaaaagtcttttgtattgggtcagg

[0748] agttgaatttggggtgggaggatggatgcaactgaagcagagtgtgggtgcccagatgtgcgctattagatgtttctctgataatgtccc

[0749] caatcataccagggagactggcattgacgagaactcaggtggaggcttgagaaggccgaaagggcccctgacctgcctggcttcctt

[0750] agcttgcccctcagctttgcaaagagccaccctaggccccagctgaccgcatgggtgtgagccagcttgagaacactaactactcaat

[0751] aaaagcgaaggtggacatgaaaaaaaaaaaaaaaaaaa

[0752] Cancer reference gene sequence

[0753] SEQ ID NO:1(ALAS1; NM_000688)

[0754] aggctgctcccggacaagggcaacgagcgtttcgtttggacttctcgacttgagtgcccgcctccttcgccgccgcctctgcagtcctc

[0755] agcgcagttatgcccagttcttcccgctgtggggacacgaccacggaggaatccttgcttcagggactcgggaccctgctggacccct

[0756] tcctcgggtttaggggatgtggggaccaggagaaagtcaggatccctaagagtcttccctgcctggatggatgagtggcttcttctcca

[0757] cctagattctttccacaggagccagcatacttcctgaacatggagagtgttgttcgccgctgcccattcttatcccgagtcccccaggcct

[0758] ttctgcagaaagcaggcaaatctctgttgttctatgcccaaaactgccccaagatgatggaagttggggccaagccagcccctcgggc

[0759] attgtccactgcagcagtacactaccaacagatcaaagaaacccctccggccagtgagaaagacaaaactgctaaggccaaggtcca

[0760] acagactcctgatggatcccagcagagtccagatggcacacagcttccgtctggacaccccttgcctgccacaagccagggcactgc

[0761] aagcaaatgccctttcctggcagcacagatgaatcagagaggcagcagtgtcttctgcaaagccagtcttgagcttcaggaggatgtg

[0762] caggaaatgaatgccgtgaggaaagaggttgctgaaacctcagcaggccccagtgtggttagtgtgaaaaccgatggaggggatcc

[0763] cagtggactgctgaagaacttccaggacatcatgcaaaagcaaagaccagaaagagtgtctcatcttcttcaagataacttgccaaaat

[0764] ctgtttccacttttcagtatgatcgtttctttgagaaaaaaattgatgagaaaaagaatgaccacacctatcgagtttttaaaactgtgaacc

[0765] ggcgagcacacatcttccccatggcagatgactattcagactccctcatcaccaaaaagcaagtgtcagtctggtgcagtaatgactac

[0766] ctaggaatgagtcgccacccacgggtgtgtggggcagttatggacactttgaaacaacatggtgctggggcaggtggtactagaaat

[0767] atttctggaactagtaaattccatgtggacttagagcgggagctggcagacctccatgggaaagatgccgcactcttgttttcctcgtgct

[0768] ttgtggccaatgactcaaccctcttcaccctggctaagatgatgccaggctgtgagatttactctgattctgggaaccatgcctccatgat

[0769] ccaagggattcgaaacagccgagtgccaaagtacatcttccgccacaatgatgtcagccacctcagagaactgctgcaaagatctga

[0770] cccctcagtccccaagattgtggcatttgaaactgtccattcaatggatggggcggtgtgcccactggaagagctgtgtgatgtggccc

[0771] atgagtttggagcaatcaccttcgtggatgaggtccacgcagtggggctttatggggctcgaggcggagggattggggatcgggatg

[0772] gagtcatgccaaaaatggacatcatttctggaacacttggcaaagcctttggttgtgttggagggtacatcgccagcacgagttctctga

[0773] ttgacaccgtacggtcctatgctgctggcttcatcttcaccacctctctgccacccatgctgctggctggagccctggagtctgtgcgga

[0774] tcctgaagagcgctgagggacgggtgcttcgccgccagcaccagcgcaacgtcaaactcatgagacagatgctaatggatgccggc

[0775] ctccctgttgtccactgccccagccacatcatccctgtgcgggttgcagatgctgctaaaaacacagaagtctgtgatgaactaatgag

[0776] cagacataacatctacgtgcaagcaatcaattaccctacggtgccccggggagaagagctcctacggattgcccccacccctcacca

[0777] cacaccccagatgatgaactacttccttgagaatctgctagtcacatggaagcaagtggggctggaactgaagcctcattcctcagctg

[0778] agtgcaacttctgcaggaggccactgcattttgaagtgatgagtgaaagagagaagtcctatttctcaggcttgagcaagttggtatctg

[0779] ctcaggcctgagcatgacctcaattatttcacttaaccccaggccattatcatatccagatggtcttcagagttgtctttatatgtgaattaa

[0780] gttatattaaattttaatctatagtaaaaacatagtcctggaaataaattcttgcttaaatggtg

[0781] SEQ ID NO:13(MMADHC;NM_015702)

[0782] cctttgcctgctcaccgccagcgtaggtgctaccaccgctgccgtcgccgccgccattttgatggcaggaagagtccggttctgggac

[0783] agctggagacagtggtggtgactgaaataactttaccaaaggaaagctattttgcgaactatcttctccagcggagatggccaatgtgct

[0784] ttgtaacagagccagactggtttcctatctcccaggattttgctctttagttaaaagggttgtcaatcccaaagccttttcgactgcaggatc

[0785] atcaggttcggatgagtctcatgtggctgctgcacctccagatatatgctctcgaacagtgtggcctgatgaaactatgggaccctttgg

[0786] acctcaagatcagaggttccagcttcctgggaacataggttttgattgtcacctcaatgggactgcttcacagaagaaaagcctggttca

[0787] taaaactttgcctgatgttctagcagaacctttatcaagtgaaagacatgagtttgtgatggcacaatatgtgaatgaatttcagggtaatg

[0788] atgcacctgttgaacaagaaattaacagtgcagaaacttactttgaaagtgccagagtagagtgtgcaatacagacatgtccagaattg

[0789] ctgcgaaaagattttgaatcactgtttccagaagtagctaatggcaaactaatgattctgactgtaacacaaaaaactaagaatgatatga

[0790] ctgtttggagtgaagaagtagaaattgaaagagaagtgctcttagaaaagttcatcaatggtgctaaggaaatttgctatgctcttcgagc

[0791] tgagggttattgggctgactttattgacccatcatctggtttggcattttttggaccatatacaaacaacactctttttgaaactgatgaacgc

[0792] taccgacatttaggattctctgttgatgaccttggatgctgtaaagtgattcgtcatagtctctggggtacccatgtagttgtagggagtatc

[0793] ttcactaatgcaacaccagacagccatattatgaagaaattaagtggaaattagcagaaatatccattcatttgctgtactatttgtatgtaa

[0794] tatttgggttgatctataaacactgtcagactaaagtttttaaaatatatttatttctaagtatttatttcagcatttatgaatttacaacattggca

[0795] agtgatttgggattttaaaattgcaaatgttcatttattcatatcattgaatacacgttgaacacatccacattgtataggatgtggtaattagc

[0796] ttgtaaccagggtatgatctgctattgttatttctcctctttattggaaaaaggcctcagttttaattattttcttcccaaaataaatcacacattt

[0797] ggttacaa

[0798] SEQ ID NO:14(PPP2CA;NM_002715)

[0799] acagagagccgagctctggagcctcagcgagcggaggaggaggcgcagcggccgacggccgagtactgcggtgagagccagc

[0800] gggccagcgccagcctcaacagccgccagaagtacacgaggaaccggcggcggcgtgtgcgtgtaggcccgtgtgcgggcggc

[0801] ggcgcgggagcagcgcggagcggcagccggctggggcgggtggcatcatggacgagaaggtgttcaccaaggagctggaccag

[0802] tggatcgagcagctgaacgagtgcaagcagctgtccgagtcccaggtcaagagcctctgcgagaaggctaaagaaatcctgacaaa

[0803] agaatccaacgtgcaagaggttcgatgtccagttactgtctgtggagatgtgcatgggcaatttcatgatctcatggaactgtttagaatt

[0804] ggtggcaaatcaccagatacaaattacttgtttatgggagattatgttgacagaggatattattcagttgaaacagttacactgcttgtagc

[0805] tcttaaggttcgttaccgtgaacgcatcaccattcttcgagggaatcatgagagcagacagatcacacaagtttatggtttctatgatgaat

[0806] gtttaagaaaatatggaaatgcaaatgtttggaaatattttacagatctttttgactatcttcctctcactgccttggtggatgggcagatctt

[0807] ctgtctacatggtggtctctcgccatctatagatacactggatcatatcagagcacttgatcgcctacaagaagttccccatgagggtcca

[0808] atgtgtgacttgctgtggtcagatccagatgaccgtggtggttggggtatatctcctcgaggagctggttacacctttgggcaagatattt

[0809] ctgagacatttaatcatgccaatggcctcacgttggtgtctagagctcaccagctagtgatggaggggatataactggtgccatgaccgg

[0810] aatgtagtaacgattttcagtgctccaaactattgttatcgttgtggtaaccaagctgcaatcatggaacttgacgatactctaaaatactct

[0811] ttcttgcagtttgacccagcacctcgtagaggcgagccacatgttactcgtcgtaccccagactacttcctgtaatgaaattttaaacttgt

[0812] acagtattgccatgaaccatatatcgacctaatggaaatgggaagagcaacagtaactccaaagtgtcagaaaatagttaacattcaaa

[0813] aaacttgttttcacatggaccaaaagatgtgccatataaaaatacaaagcctcttgtcatcaacagccgtgaccactttagaatgaacca

[0814] gttcattgcatgctgaagcgacattgttggtcaagaaaccagtttctggcatagcgctatttgtagttacttttgctttctctgagagactgc

[0815] agataataagatgtaaacattaacacctcgtgaatacaatttaacttccatttagctatagctttactcagcatgactgtagataaggatagc

[0816] agcaaacaatcattggagcttaatgaacatttttaaaaataattaccaaggcctcccttctacttgtgagttttgaaattgttctttttattttca

[0817] gggataccgtttaatttaattatatgatttgtctgcactcagtttattccctactcaaatctcagccccatgttgttctttgttattgtcagaacct

[0818] ggtgagttgttttgaacagaactgttttttccccttcctgtaagacgatgtgactgcacaagagcactgcagtgtttttcataataaacttgtg

[0819] aactaagaactgagaaggtcaaattttaattgtatcaatgggcaagactggtgctgtttattaaaaaagttaaatcaattgagtaaattttag

[0820] aatttgtagacttgtaggtaaaataaaaatcaagggcactacataacctctctggtaactccttgacattcttcagattaacttcaggatttat

[0821] ttgtatttcacatattacaatttgtcacattgttggtgtgcactttgtgggttcttcctgcatattaacttgtttgtaagaaaggaaatctgtgct

[0822] gcttcagtaagacttaattgtaaaaccatataacttgagatttaagtctttgggttttgttttaataaaacagcatgttttcaggtagagcttaa

[0823] actaaatgatgtgtttacttagtgcagtttctggttatgaatattatattgctatgtgtatattatatggactctttaaaatgattgacagattggc

[0824] aaattcttaaatctttgtacattgttgagtcatatgttcttagagttaaatttgtctcagataagaaagtgttaaagcattagcctgtgtcaagtt

[0825] ctttgagtgatactagtgaaaccaaatagaaaactattgttggatcatgatttagtcttatgtacattcacccgaagacaaaaatggtactta

[0826] aagtggcagtgttcaacatttaatgagtttttccccttttatccttcgaataggattagatgtttaaaaaaaagttcttctgtgggaactaatat

[0827] ttgatattttaacctaccagagtaaacaggaacacttaatcatacttgtgagtgtagtaaataaaagttttcttgctttgtgctgtgttgaatct

[0828] ggaaccaacagggaagttatagcatatcccctttctaaaatgcttgaggaacacatacataccgaatgtcttttctgatctaattgatagta

[0829] tttttagtggcttgtggagttaattttccaaagcaaaaggccattagggtttctacatttcatttcatttcattcttttctttcacaagaaatacatt

[0830] ctctgtgtgtctttttgttgctctgtcactctatgccctttctctccgactgaacaaatagcttatccatgtgcagtggttttaatacccaaaca

[0831] atctagacaccaagcagctattttttccggtcctgatatcagaattgaccaaggaatacgtatattgtaattgacacgtggtggtatcttc

[0832] caggtacaaattctctaaattttgtggttagcagaatgggacttgtgataagaataagcttggttttagcataactagggtttaaaataattgtt

[0833] taattatagactgaccgggctttggccatctaaactggaagtgttagtaccctaccttctttgaaaatggctatggtaaggaaaagt

[0834] gttagtaaattatgtattttcttgaaaaatacataattatggttggatgggaatcactaagttgggtgttaactgatgtctcaattagtaacatt

[0835] aggattttcattaataaacctaaaaagctttccctaagaacaggcctggcacagtggctcatgcctgtaatcccagcactttgggagacc

[0836] agggtgggtggatcccttgaggtcaggagttgaagagcagcctggccaacatggcaaaaccccatctctacaaaaaatatgaaaatc

[0837] agccgggcttggtggcataccatagtcccagctacttgggaggctgaggtgagaggatcgcctgagcccaggagatggaggttgca

[0838] gtgggctgagattgtgccactgtactccagtctgggtgacagagccagaccctgtctcaaaaataaagaggattctgagtttgtatagtg

[0839] agggcttgcagaaattttgaaacttattttgtaagtttacaatgaatttgtacatgatgtgctcatgtcttgggttgagtatcctagacatgatt

[0840] ttttcatttgctgcatattaaacatttgttggttgtagtcggtatttcttaaatagaagtttgtcaatattagattagtttcaagaaggacttagct

[0841] caggaaaaggatagttatttctgtggttctcagctttgatgcctacagagattctgatttaaattgctggaggagggcccaggcatacattt

[0842] tttaagtttcccaagtgattctaatttacatctagggttaaaaacagccaggcgcggtggctcatgcctgtaatcccagcacttggggag

[0843] gccggggtgggtggatcacctgatgtcaggagatcgagaccagcctggccagcgtggtgaaaccccatctctattaaaaatacaaaa

[0844] ctgaccggtgtggtggcaggagcctgtaatcccagctacttgggaggctgaggcaggagaatcacttgaactcaggaggaggaggt

[0845] tacagtgagcccagatcgtgccattgcactccagcctgggcaaaaagagcaaaactccatctcaaaaaacaaaaacacctcactgctg

[0846] tttcctaagtacatacttaagaaaaattgggatacatggtggtggttcatggatgttgataaggaattaaaatgtaccgtgcgactctctgttt

[0847] cagtggtgacttttacctgtttagtataaatattcctttgcttccaaccataaatgtgttcttagaaatgggcctatagtttagtaacctatagtt

[0848] tggtaataggcttgtttgttttcagatggattttggttctgtgagctaaagctattttgcattaaagccttcgtcctcaca

[0849] SEQ ID NO:15(TFRC;NM_003234)

[0850] agagcgtcgggatatcgggtggcggctcgggacggaggacgcgctagtgtgagtgcgggcttctagaactacaccgaccctcgtgt

[0851] cctcccttcatcctgcggggctggctggagcggccgctccggtgctgtccagcagccatagggagccgcacggggagcgggaaag

[0852] cggtcgcggccccaggcggggcggccgggatggagcggggccgcgagcctgtggggaaggggctgtggcggcgcctcgagcg

[0853] gctgcaggttcttctgtgtggcagttcagaatgatggatcaagctagatcagcattctctaacttgtttggtggagaaccattgtcatatac

[0854] ccggttcagcctggctcggcaagtagatggcgataacagtcatgtggagatgaaacttgctgtagatgaagaagaaaatgctgacaat

[0855] aacacaaaggccaatgtcacaaaaccaaaaaggtgtagtggaagtatctgctatgggactattgctgtgatcgtctttttcttgattggatt

[0856] tatgattggctacttgggctattgtaaaggggtagaaccaaaaactgagtgtgagagactggcaggaaccgagtctccagtgaggga

[0857] ggagccaggagaggacttccctgcagcacgtcgcttatattgggatgacctgaagagaaagttgtcggagaaactggacagcacag

[0858] acttcaccggcaccatcaagctgctgaatgaaaattcatatgtccctcgtgaggctggatctcaaaaagatgaaaatcttgcgttgtatgt

[0859] tgaaaatcaatttcgtgaatttaaactcagcaaagtctggcgtgatcaacattttgttaagattcaggtcaaagacagcgctcaaaactcg

[0860] gtgatcatagttgataagaacggtagacttgtttacctggtggagaatcctgggggttatgtggcgtatagtaaggctgcaacagttact

[0861] ggtaaactggtccatgctaattttggtactaaaaaagattttgaggatttatacactcctgtgaatggatctatagtgattgtcagagcagg

[0862] gaaaatcacctttgcagaaaaggttgcaaatgctgaaagcttaaatgcaattggtgtgttgatatacatggaccagactaaatttcccatt

[0863] gttaacgcagaactttcattctttggacatgctcatctggggacaggtgacccttacacacctggattcccttccttcaatcacactcagttt

[0864] ccaccatctcggtcatcaggattgcctaatatacctgtccagacaatctccagagctgctgcagaaaagctgtttgggaatatggaagg

[0865] agactgtccctctgactggaaaacagactctacatgtaggatggtaacctcagaaagcaagaatgtgaagctcactgtgagcaatgtg

[0866] ctgaaagagataaaaattcttaacatctttggagttattaaaggctttgtagaaccagatcactatgttgtagttggggcccagagagatg

[0867] catggggccctggagctgcaaaatccggtgtaggcacagctctcctattgaaacttgcccagatgttctcagatatggtcttaaaagatg

[0868] ggtttcagcccagcagaagcattatctttgccagttggagtgctggagactttggatcggttggtgccactgaatggctagagggatac

[0869] ctttcgtccctgcatttaaaggctttcacttatattaatctggataaagcggttcttggtaccagcaacttcaaggtttctgccagcccactgt

[0870] tgtatacgcttattgagaaaacaatgcaaaatgtgaagcatccggttactgggcaatttctatatcaggacagcaactgggccagcaaa

[0871] gttgagaaactcactttagacaatgctgctttccctttccttgcatattctggaatcccagcagtttctttctgtttttgcgaggacacagatta

[0872] tccttatttgggtaccaccatggacacctataaggaactgattgagaggattcctgagttgaacaaagtggcacgagcagctgcagag

[0873] gtcgctggtcagttcgtgattaaactaacccatgatgttgaattgaacctggactatgagaggtacaacagccaactgctttcatttgtga

[0874] gggatctgaaccaatacagagcagacataaaggaaatgggcctgagtttacagtggctgtattctgctcgtggagacttcttccgtgcta

[0875] cttccagactaacaacagatttcgggaatgctgagaaaacagacagatttgtcatgaagaaactcaatgatcgtgtcatgagagtggag

[0876] tatcacttcctctctccctacgtatctccaaaagagtctcctttccgacatgtcttctggggctccggctctcacacgctgccagctttactg

[0877] gagaacttgaaactgcgtaaacaaaataacggtgcttttaatgaaacgctgttcagaaaccagttggctctagctacttggactattcag

[0878] ggagctgcaaatgccctctctggtgacgtttgggacattgacaatgagttttaaatgtgatacccatagcttccatgagaacagcagggt

[0879] agtctggtttctagacttgtgctgatcgtgctaaattttcagtagggctacaaaacctgatgttaaaattccatcccatcatcttggtactact

[0880] agatgtctttaggcagcagcttttaatacagggtagataacctgtacttcaagttaaagtgaataaccacttaaaaaatgtccatgatgga

[0881] atattcccctatctctagaattttaagtgctttgtaatgggaactgcctctttcctgttgttgttaatgaaaatgtcagaaaccagttatgtgaa

[0882] tgatctctctgaatcctaagggctggtctctgctgaaggttgtaagtggtcgcttactttgagtgatcctccaacttcatttgatgctaaata

[0883] ggagataccaggttgaaagaccttctccaaatgagatctaagcctttccataaggaatgtagctggtttcctcattcctgaaagaaacagt

[0884] taactttcagaagagatgggcttgttttcttgccaatgaggtctgaaatggaggtccttctgctggataaatgaggttcaactgttgattg

[0885] caggaataaggccttaatatgttaacctcagtgtcatttatgaaaagaggggaccagaagccaaagacttagtatattttcttttcctctgt

[0886] ccctcccccataagcctccatttagttctttgttatttttgtttcttccaaagcacattgaaagagaaccagtttcaggtgtttagttgcagac

[0887] tcagtttgtcagactttaaagaataatatgctgccaaattttggccaaagtgttaatcttaggggagagctttctgtccttttggcactgagat

[0888] atttattgtttatttatcagtgacagagttcactataaatggtgtttttttaatagaatataattatcggaagcagtgccttccataattatgaca

[0889] gttatactgtcggttttttttaaataaaagcagcatctgctaataaaacccaacagatactggaagttttgcatttatggtcaacacttaagg

[0890] gttttagaaaacagccgtcagccaaatgtaattgaataaagttgaagctaagatttagagatgaattaaatttaattaattaggggttgctaagaa

[0891] gcgagcactgaccagataagaatgctggttttcctaaatgcagtgaattgtgaccaagttataaatcaatgtcacttaaaggctgtggtag

[0892] tactcctgcaaaattttatagctcagtttatccaaggtgtaactctaattcccattttgcaaaatttccagtacctttgtcacaatcctaacaca

[0893] ttatcgggagcagtgtcttccataatgtataaagaacaaggtagtttttacctaccacagtgtctgtatcggagacagtgatctccatatgtt

[0894] acactaagggtgtaagtaattatcgggaacagtgtttcccataattttcttcatgcaatgacatcttcaaagcttgaagatcgttagtatcta

[0895] acatgtatcccaactcctataattccctatcttttagttttagttgcagaaacattttgtggtcattaagcattgggtgggtaaattcaaccact

[0896] gtaaaatgaaattactacaaaatttgaaatttagcttgggtttttgttacctttatggtttctccaggtcctctacttaatgagatagtagcatac

[0897] atttataatgtttgctattgacaagtcattttaactttatcacattatttgcatgttacctcctataaacttagtgcggacaagttttaatccagaa

[0898] ttgaccttttgacttaaagcagagggactttgtatagaaggtttgggggctgtggggaaggagagtcccctgaaggtctgacacgtctg

[0899] cctacccatcgtggtgatcaattaaatgtaggtatgaataagttcgaagctccgtgagtgaaccatcattataaacgtgatgatcagctgt

[0900] ttgtcatagggcagttggaaacggcctcctagggaaaagttcatagggtctcttcaggttcttagtgtcacttacctagatttcagcctca

[0901] cttgaatgtgtcactactcacagtctctttaatcttcagttttatctttaatctcctcttttatcttggactgacatttagcgtagctaagtgaaaa

[0902] ggtcatagctgagattcctggttcgggtgttacgcacacgtacttaaatgaaagcatgtggcatgttcatcgtataacacaatatgaatac

[0903] agggcatgcattttgcagcagtgagtctcttcagaaaacccttttctacagttagggttgagttacttcctatcaagccagtacgtgctaac

[0904] aggctcaatattcctgaatgaaatcagactagtgacaagctcctggtcttgagatgtcttctcgttaaggagatgggccttttggaggt

[0905] aaaggataaaatgaatgagttctgtcatgattcactattctagaacttgcatgacctttactgtgttagctctttgaatgttcttgaaattttag

[0906] actttctttgtaaaaatgatatgtccttatcattgtataaaagctgttatgtgcaacgtgtggagattccttgtctgatttaataaataactt

[0907] aaacactgaaaaaaaaaa

[0908] SEQ ID NO:16(CLTC:NM_004859)

[0909] agccgtttccggagtctgcgctgcccggttcgccattgcggctctctggccctggagcctcgccccgacccgagctctttc

[0910] gtctgcctgccagtttcctgcgtccccggagaggatctgctgagcccagccccccctctctctctctccttggagagc

[0911] gggcagccactgccccgcagccccagtgacaggagagaccataacccccgacagcgccatggcccagattgccaattcgtttt

[0912] caggagcatctccagctccagaacctgggtatcaacccagcaaacattggcttcagtaccctgactatggagtctgacaaattcatctg

[0913] cattagagaaaaagtaggagagcaggcccaggtggtaatcattgatatgaatgacccaagtaatccaattcgaagaccaatttcagca

[0914] gacagcgccatcatgaatccagctagcaaagtaattgcactgaaagctgggaaaactcttcagatttttaacattgaaatgaaaagtaaa

[0915] atgaaggctcataccatgactgatgatgtcaccttttggaaatggatctctttgaatacggttgctcttgttacggataatgcagtttatcact

[0916] ggagtatggaaggagagtctcagccagtgaaaatgtttgatcgccattctagccttgcagggtgccagattatcaattaccgtacagatg

[0917] caaaacaaaagtggttacttctgactggtatatctgcacagcaaaatcgtgtggtgggagctatgcagctatattctgtagataggaaag

[0918] tgtctcagcccattgaaggacatgcagctagctttgcacagtttaagatggaaggaaatgcagaagaatcaacgttattttgttttgcagtt

[0919] cggggccaagctggagggaagttacatattattgaagttggcacaccacctacagggaaccagccctttccaaagaaggcagtggat

[0920] gtcttctttcctccagaagcacaaaatgattttcctgttgcaatgcagatcagtgaaaagcatgatgtggtgttcttgataaccaagtatggt

[0921] tatatccacctctatgatcttgagactggtacctgcatctacatgaatagaatcagtggagaaacaatttttgttactgcacctcatgaagc

[0922] cacagctggaataattggagtaaacagaaagggacaagttctgtcagtgtgtgtggaagaagaaaacataattccttacatcaccaatg

[0923] ttctacaaaatcctgatttggctctgagaatggctgtacgtaataacttagccggtgctgaagaactctttgcccggaaatttaatgctcttt

[0924] ttgcccagggaaattactcggaggcagcaaaggtggctgctaatgcaccaaagggaattcttcgtactccagacactatccgtcggttc

[0925] cagagtgtcccagcccagccaggtcaaacttctcctctacttcagtactttggtatccttttggaccagggacagctcaacaaatacgaa

[0926] tccttagagctttgtaggcctgtacttcagcaagggcgaaaacagcttttggagaaatggttaaaagaagataagctggaatgttctgaa

[0927] gaactgggtgatcttgtgaaatctgtggaccctacattggcacttagtgtgtacctaagggctaacgtcccaaataaagtcattcagtgct

[0928] ttgcagaaacaggtcaagtccaaaagattgttttatatgctaaaaaagttggatacactccagattggatatttctgctgagaaatgtaatg

[0929] cgaatcagtccagatcagggacagcagtttgcccaaatgttagttcaagatgaagagcctcttgctgacatcacacagattgtagatgtc

[0930] tttatggaatacaatctaattcagcagtgtactgcattcttgcttgatgctctgaagaataatcgcccatctgaaggtcctttacagacgcg

[0931] gttacttgagatgaaccttatgcatgcgcctcaagttgcagatgctattctaggcaatcagatgttcacacattatgaccgggctcatattg

[0932] ctcaactgtgtgaaaaggctggcctactgcagcgtgcattagaacatttcactgatttatatgatataaaacgtgcagtggttcacaccca

[0933] tcttcttaaccctgagtggttagtcaactactttggttccttatcagtagaagactccctagaatgtctcagagccatgctgtctgccaacat

[0934] ccgtcagaatctgcagatttgtgttcaggtggcttctaaatatcatgaacaactgtcaactcagtctctgattgaactttttgaatctttcaag

[0935] agttttgaaggtctcttttattttctgggatccattgttaactttagccaggacccagatgtgcactttaaatatattcaggcagcttgcaaga

[0936] ctgggcaaatcaaagaagtagaaagaatctgtagagaaagcaactgctacgatcctgagcgagtcaagaattttcttaaggaagcaaa

[0937] actaacagatcagctaccacttatcattgtgtgtgatcgatttgactttgtccatgatttggtgctctatttatatagaaataatcttcaaaagt

[0938] atatagagatatatgtacagaaggtgaatccaagtcgacttcctgtagttattggaggattacttgatgttgactgttctgaagatgtcataa

[0939] aaaacttgattcttgttgtaagaggtcaattctctactgatgagcttgttgctgaggttgaaaaaagaaacagattgaaactgcttctgcctt

[0940] ggctagaggccagaattcatgagggctgtgaggagcctgctactcacaatgccttagccaaaatctacatagacagtaataacaaccc

[0941] ggagagatttcttcgtgaaaatccctactatgacagtcgcgttgttggaaagtattgtgagaagagagatccacatctggcctgtgttgct

[0942] tatgaacgtggccaatgtgatctggaacttattaatgtttgcaatgagaattccctcttcaaaagtctttctcgctacctggtacgtcgaaag

[0943] gatccagaattgtggggcagcgtgctgctggaaagcaatccttacaggagacccctaattgaccaggttgtacaaacagctttgtctga

[0944] gactcagccctgaagaagtgtcagtaactgtaaaggctttcatgactgcagaccttcctaatgaactcattgaactgctggagaaaat

[0945] tgtccttgataactctgtattcagtgaacacaggaatctgcaaaacctccttatcctcactgcaattaaggctgaccgtacacgtgttatgg

[0946] agtatattaaccgcctggataattatgatgccccagatattgccaatatcgccatcagcaatgagctgtttgaagaagcatttgccattttc

[0947] cggaaatttgatgtcaatacttcagcagttcaggtcttaattgagcatattggaaacttggatcgggcatatgagtttgctgaacgttgcaa

[0948] tgaacctgcggtctggagtcaacttgcaaaagcccagttgcagaaaggaatggtgaaagaagccattgattctttatcaaagcagatg

[0949] atccttctccctacatggaagttgttcaggctgccaatactagtggaaactgggaagaactggtgaagtacttgcagatggcccgtaag

[0950] aaggctcgagagtcctatgtggagacagaactgatattcgcactggctaaaacaaaccgccttgcagagttagaagaatttatcaatgg

[0951] accaaataatgctcatatccaacaagttggtgaccgttgttatgatgaaaaaatgtatgatgctgctaagttgttgtacaataatgtttccaa

[0952] ttttggacgtttggcatctaccctggttcacctgggtgaatatcaggcagctgttgatggggctaggaaagctaacagtactcgaacatg

[0953] gaaagaggtctgcttcgcctgtgtagatgggaaagaattccgtcttgctcagatgtgtggacttcatattgttgtacatgcagatgaattag

[0954] aagaacttatcaactactatcaggatcgtggctattttgaagagctgatcaccatgttggaagcagcactgggacttgagcgagctcaca

[0955] tgggaatgtttactgaattagctattctatactctaaatttaagcctcagaaaatgagggagcacctggagctgttctggtctagagtgaat

[0956] attcccaaggtgctaagagctgcagaacaagctcatctttgggcagaactggtgttttgtatgacaagtatgaagaatatgataatgcca

[0957] taattaccatgatgaatcatccaactgatgcctggaaagaagggcaattcaaagatatcattaccaaggttgccaatgtggaactatact

[0958] acagagcaatacagttctacttagaattcaagcctctgttgttaaatgatttgctgatggtgctgtctccacggttggatcacactcgtgca

[0959] gtcaattattcagcaaggttaaacagctaccactggtgaaaccgtatttgcgttcagttcagaaccataacaacaaatctgtgaatgaat

[0960] cattgaacaatctttttattacagaagaagattatcaggctctgcgaacatcaatagatgcttatgacaactttgacaatatctcgcttgctc

[0961] agcgtttggaaaaacatgaactcattgagttcaggagaattgctgcttatctcttcaaaggcaacaatcgctggaaacagagtgtagagc

[0962] tgtgcaagaaagacagcctttacaaggatgcaatgcagtatgcttctgaatctaaagatactgaattggctgaagaactcctgcagtggt

[0963] ttttgcaggaagaaaaaagagagtgctttggagcttgtctgtttacctgttacgatcttttaaggccagatgtcgtcctagaaactgcatgg

[0964] aggcacaatatcatggattttgccatgccctatttcatccaggtcatgaaggagtacttgacaaaggtggataaattagatgcttcagaat

[0965] cactgagaaaagaagaagaacaagctacagagacacaacccattgtttatggtcagccccagttgatgctgacagcaggacccagtg

[0966] ttgccgtccctccccaggcaccttttggttatggttataccgcaccaccgtatggacagccacagcctggctttgggtacagcatgtgag

[0967] atgaagcgctgatcctgtagtcacctattttcgtactgaaacatcgtctttacccacttctcagtttataatgggggaaaacaggcaacgtg

[0968] ttcttgtaacctttatttcatgaaggacttctttttgtttctaactataaacttggatcacctatgttaaaaccttatttcacattccacatcatttta

[0969] gaatttattttcgaaggggaatagtttcaatgttttattcacttgggctttttttcttccccctctttctttaaagaactgctcaatattcaatctgtt

[0970] gtgaagaacctgatttgcactctgtagtgtttaaagaaacaaagaaactctaatattgaatctcttaaatttagtgtatgtaaacagcttaca

[0971] aatacgtattgtctaaatgcatttaaatctgttttattcaaagaaaagctaaagcaaaaacactggcatatgaccatgcaagactgtcagtg

[0972] ccaacaaagacaacactaatcagcacatcgtacactggattgcagtgcttcccagattattgaaaaatgttacagacaacttgcctgattt

[0973] ttaaatgagcgtaaaaggccctctaacctatgcaggtttccccattatgcatatagaaaatgctagtatgttttgctcacttcatatgtaaca

[0974] ggtgccccttatgttgtgctgtatcctgtgctttttctgtgggaccattccattcaggagcaaagagcaccatgattccaatcttgtgtgtgttt

[0975] actaacccttccctgaggtttgtgtatgttggatattgtggtgttttagatcactgagtgtacagaagagagaaattcaaacaaaatattgct

[0976] gttcttcagttttgtttgtggaatttgaaattactcaaatttaaaataaattactggactgtggaaataacatagaattgaagttttaattaaata

[0977] ccactcaaacgaaaagaacagtagtttttgtagttttatattggatactgaggcattagggaggcatgaaaggaagaggaatgaggatt

[0978] gagacatgtgaagacattgtgcattatatcaatgtgcattcctgtagttcattaacaaggtacatgcaatagtctaaagaaccagagtcac

[0979] tactatagtggcttaacatttaatctgtctccaatattttaaccaagtgacaccgaggtttttatcgaagcatttcacttaaatgaacaaatcat

[0980] ggctgttatattaacttgaaataaaatatatttaaacatgtagttaacatgctctttctcatcatgaattataaaggtgcccaaattatgggtat

[0981] cttgatttttttcaaaaattggttgtgtgaatccagctcttgcttatggctgagaatgttcctaaagttcatttcactatcccttaatcctcaata

[0982] gctacacatacccccttgcctagttagtcttgcactgctttacaactctcttaggcaaattacctaacataagctccacagaagagtttttta

[0983] aaacttaaaatccctgtcccaccaggtgtggtggctcatgcctgtaatcccagcactttgggaggctgaggcaggcggattgcttgagg

[0984] ccaggagttcgagacaagcctgaccaacatggtgactccatctctactaaaaataaaaaatcagccaggtgtgatggtggatgcctata

[0985] agatcctagctgctactcaggaggcaggaggctgaggcatgagaatcacttgaacctgtgaggcaaaggttgtagtgagccaagatc

[0986] acgtcactgcactccatcctgggtgacagaatgagactcttgtctcaaaaaacaaaaaaacctatttagtttcatccaggacattaagtga

[0987] aagtgtttaatgtgagtgcaggtcagtgaagaatacaatggtgcccctgctttgatttgtgggaggtgccatcagttttccccaccactgc

[0988] ttttgcaccctcagtgcaaatatcaacacagagaaaaaaaaaataataatagtattatgaaaacattgactctgtggagaccctgtgggtt

[0989] ctgcagagtatactttgaaaactataagattatgagttctataaaataccaagtacagatgaactagaaaacaagcatatatccacaatta

[0990] ccagaattttcatagtattccttttagtagatatgccatgacagttacttagccaccccatgccatcctgttgggaatgccactacctttgaat

[0991] ttggagccatcagtctatctgaggcaaacctcagaaattacctgagccagaatcatgcacttagcactgagttcaactcagtcttaacgtt

[0992] gcattttgttgcatcacttttgcagccagtaaacattctgtttagaaatagttgctattgtggcatcattcacagttgcaatttttctcctgtgct

[0993] gtcatatgcctgtgcgaagtgtattaaatgaatcaaatattagaaatttcaaggctgtctttagtgttctgcccacaatactccaccaatgac

[0994] cttagaccttgagatggtgctataacgctgcttatattgcagttaaccatagagaggtggagccacttaaaatgggcctattaccaaaatt

[0995] ctccctggcatcattacagttcttggggatacactgattttagaaattactgtcagtatcctcctaaaagaaggcttaaatagtttctaacaa

[0996] gatgactatcaggaagctatgtggcttggggagttgggaccctctgcctcatattctgggtaactggtataacatagtaattattaggattg

[0997] tatcaagtgtatctaaagatcaaaagggaaaaaaggcacccttaattgtcaaagttagttctaggaaaaaaatactagcagggatgacaa

[0998] actacgttcactttacagttaccataaattcttacttgggcccacccatttccatctgttaaggtccataaggttactcaagtttcagtggctt

[0999] cattgttacaagaaaggggggaagtgagaaactttaataggctgtgatttagaaccaactgctgattaacctttggccagtatgggtcagg

[1000] gaaggtgaacatgaaacactgcacagtaggttgtatcaatgctataaaac

[1001] Statistical analysis

[1002] The Cancer Genome Atlas (TCGA) prostate cancer dataset (1) was used to identify a gene group with prognostic significance for aggressive prostate cancer. The TCGA dataset contains RNA sequencing data (approximately 21,000 genes) and clinical data from 399 samples. Eleven gene groups were identified using three methods:

[1003] 1) Statistical modeling to identify genes that increase statistically significant prognostic values ​​based on standard clinical information currently used by clinicians.

[1004] 2) ARACNe: Gene regulatory network analysis to identify genes regulating the prognostic genes identified in (1) above.

[1005] 3) Analysis of bioinformatics databases (originality approach analysis: databases used for annotation, visualization and integration of findings to identify genes from (1) and / or (2) above that have cancer-related biological functions).

[1006] The two clinical outcomes were further evaluated by 2-fold cross-validation of all 2,047 possible combinations of genes 1 to 11 in a sample cohort from Ireland (PCRC, N = 426 samples). Eleven gene groups (i.e., TTC21B, MCM3, MTUS1, FOXM1, CUL4A, MCM3, GPC1, PCNA, IRF7, BRIP1, ULK1, and SPAG5) through 100 iterations.

[1007] The primary clinical outcome was adverse pathology (AP), defined as any of the following six characteristics: pathological Gleason grade ≥4+3, pathological stage ≥T3, positive nodule, presence of grade III Gleason 5, presence of cribriform lesions, or biochemical relapse. AP outcomes were modeled using binary logistic regression, and genetics were assessed using the likelihood ratio statistic (LRS, higher values ​​are better) and the area under the receiver operating characteristic curve (AUC, values ​​can range from 0.5 to 1.0, higher values ​​are better).

[1008] The secondary clinical outcome of the study was the time to biochemical relapse (BCR). BCR outcomes were modeled using Cox proportional hazards regression, and genes were assessed using the likelihood ratio statistic (LRS, higher values ​​are better) and the C-index (similar to the AUC above, values ​​can range from 0.5 to 1.0, higher values ​​are better).

[1009] Immunohistochemistry

[1010] TMA slides were dewaxed in xylene and rehydrated in a gradient of decreasing alcohols. Heat-mediated antigen retrieval was performed for 15 minutes at 95°C using 10 mM sodium citrate buffer (pH 6.0) in a PT module (LabVision, UK). Staining was performed using a LabVision IHC kit (LabVision, UK). Endogenous peroxidase activity was blocked by incubation with 3% hydrogen peroxide for 10 minutes. Sections were blocked in UV blocking agent for 10 minutes and incubated with the associated primary antibody for 1 hour. Sections were washed in phosphate-buffered saline containing 0.1% Tween 20 (PBS-T), followed by application of the primary antibody enhancer for 20 minutes, and then washed in PBS-T. Sections were then incubated with HRP polymer for 15 minutes, washed in PBS-T, and then developed for 10 minutes using diaminobenzidine (DAB) solution (LabVision, UK). All incubation and washing phases were performed at room temperature. The sections were counterstained in hematoxylin, dehydrated in alcohol and xylene, and fixed with DPX mounting medium. As a negative control, PBS-T was used instead of the primary antibody.

[1011] The primary antibodies used are anti-FOXM1 (MilliporeSigma / MerckKGaA, catalog number SAB1412254), anti-PTTG1 (MilliporeSigma / MerckKGaA, catalog number WH0009232M1), anti-ZNF367 (MilliporeSigma / MerckKGaA, catalog number HPA015785), anti-AGER (MilliporeSigma / MerckKGaA, catalog number AV41642), anti-CUL4A (MilliporeSigma / MerckKGaA, catalog number SAB1411512), anti-TTC21B (MilliporeSigma / MerckKGaA, catalog number HPA035495), and anti-MCM3 (MilliporeSigma / MerckKGaA, catalog number HPA035495). Merck Group, catalog number SAB1412669), Anti-GPC1 (Milliborg Sigma / Merck Group, catalog number HPA030571), Anti-PCNA (Milliborg Sigma / Merck Group, catalog number HPA030521), Anti-MTUS1 (Milliborg Sigma / Merck Group, catalog number WH0057509M1), Anti-IRF7 (Milliborg Sigma / Merck Group, catalog number PRS3941), Anti-BRIP1 (Milliborg Sigma / Merck Group, catalog number HPA005474), Anti-ULK1 (Milliborg Sigma / Merck Group, catalog number WH0008408M1), and Anti-SPAG5 (Milliborg Sigma / Merck Group, catalog number HPA022479).

[1012] TMA analysis

[1013] The slides were scanned at 20x magnification using a ScanScope XT slide scanner (Aperio Technologies, CA). For manual scoring, the staining of tumor cells was assessed by a pathologist based on the following: intensity (grades negative (0), weak (1), moderate (2), and strong (3)); and percentage (range 0–6 (0 = 0–1%; 1 = 1–10%; 2 = 10–25%; 3 = 25–50%; 4 = 50–75%; 5 = 75–90%; 6 = 90–100%).

[1014] Test samples

[1015] Archived formalin-fixed, paraffin-embedded (FFPE) prostate cancer tissue samples were used for training and validation of the claimed invention. Prostate tissue was fixed in formalin and embedded in paraffin using standard protocols. Serial 5 μm sections were prepared and mounted on glass slides; for radical prostatectomy (RP), samples of ten to fourteen sections were prepared, and for biopsy samples, six sections were prepared. The first section was stained with hematoxylin and eosin (H&E), and the last slide in the series of RP samples was stained with H&E. Pathologists examined the stained sections to identify areas containing tumors. Tumor tissue was manually dissected from unstained slides using a scalpel and transferred to microcentrifuge tubes for RNA purification.

[1016] RNA purification

[1017] Total RNA was manually purified from dissected FFPE prostate tumor tissue using the RNeasy FFPE Spin-Column Kit (Qiagen) according to the manufacturer's instructions. Dewaxing was performed using xylene, followed by treatment with DNase I to produce DNA-free RNA. Nucleic acid concentration was determined spectrophotometrically at 260 nm.

[1018] Gene expression analysis of RNA samples from radical prostatectomy (RP) tissues

[1019] For RNA samples derived from RP tissue, expression of the gene groups shown in this paper was measured using one-step reverse transcription quantitative polymerase chain reaction (RT-qPCR). Hydrolysis probe gene expression assays (manufactured by Integrated DNA Technologies) at a primer:probe ratio of 3:1 and performed on MicroAmp... TM Samples were spotted in a rapid optical 96-well plate (Applied Biosystems). Following the manufacturer's instructions, TaqPath was used. TMPCR reaction mixtures were prepared using a 1-step RT-qPCR master mix (Applied Biosystems). PCR reactions were performed in 20 μL final volumes per well, containing 5 or 10 ng RNA per reaction. Thermal cycling was performed on a 7500 Rapid Dx Real-Time PCR instrument (Applied Biosystems) using SDS software v1.4.1 in rapid cycling mode with the following parameters: 25°C for 2 min, 50°C for 15 min, 95°C for 2 min, followed by 40 cycles of 95°C for 3 sec and 60°C for 30 sec. Data acquisition was performed during the annealing and extension steps at 60°C. The threshold was set to 0.1, and automated baseline was enabled for cycle-quantitative (Cq) assays. RNA samples / expression assays were tested in triplicate per run. For each hydrolysis probe assay, 10 ng / well was used to test the positive control human total reference RNA (Agilent Technologies), and RT-PCR grade water (Life Technologies) was used as the negative control in one reaction / hydrolysis probe assay / PCR plate.

[1020] Gene expression analysis of biopsy RNA samples

[1021] The expression of the gene group shown in this paper was measured using a three-step process of reverse transcription, pre-amplification, and quantitative polymerase chain reaction (RT-PA-qPCR) to enhance the sensitivity of the test for the typical low RNA yield in small prostate biopsy samples.

[1022] Reverse transcription (RT) was performed using a high-capacity cDNA reverse transcription kit with RNase inhibitors (Applied Biosystems) in a final reaction volume of 20 mL, with 25–75 ng RNA per reaction. An oligonucleotide pool containing specific reverse primers for each hydrolysis probe assay was used to initiate the RT reaction, with a final concentration of 100 nM for each primer. The RT reactions were incubated using a SimpliAmp thermal cycler (Applied Biosystems) at 25 °C for 10 min, at 37 °C for 120 min, and at 85 °C for 5 min.

[1023] use PreAmp master mix (Applied Biosystems) was used for pre-amplification (PA) PCR in a final reaction volume of 30 mL. Each PA-PCR reaction contained 6 mL of cDNA from the RT reaction and specific forward and reverse primers for each hydrolysis probe assay (shown below), at a final concentration of 37.5 nM / reaction. The PA-PCR reactions were incubated using a SimpliAmp thermal cycler (Applied Biosystems) at 95 °C for 10 min, followed by 10 cycles of 95 °C for 15 sec and 60 °C for 4 min, and finally an inactivation step at 99 °C for 10 min.

[1024] qPCR was performed using a 96-well plate via spotting of hydrolyzed probe gene expression assays as described above for the 1-step RT-qPCR method. Follow the manufacturer's instructions. The PCR reaction mixture was prepared using a rapid advanced master mix (Applied Biosystems). PCR reactions were performed in 20 μL final volumes per well, containing 0.19 mL of pre-amplified cDNA per reaction. Cycling parameters were: 40 cycles of 50°C for 2 min, 95°C for 2 min, followed by 95°C for 3 sec and 60°C for 30 sec. PCR instrumentation and threshold settings were as described above for the 1-step RT-qPCR method. In each run, the pre-amplified cDNA sample / expression assay was tested in triplicate. For each batch of test samples, the positive control human total reference RNA (Agilent Technologies) and the negative template-free control underwent the RT, PA-PCR, and qPCR steps in parallel.

[1025] result

[1026] Statistical modeling and ARACNe analysis using TCGA identified regulators of the following 11 gene groups that have statistically significant prognostic effects on invasive prostate cancer: TTC21B, MCM3, MTUS1, FOXM1, CUL4A, GPC1, PCNA, IRF7, BRIP3, ULK1, and SPAG5. Analysis of bioinformatics databases confirmed that these genes are involved in cancer-related biological processes.

[1027] In the second sample cohort (PCRC, N = 426 samples), two clinical outcomes for the 11 gene groups were further evaluated by performing 2-fold cross-validation for 100 iterations on all 2,047 possible combinations of genes 1 to 11.

[1028] Results of all 2,047 gene combinations

[1029] For the main clinical outcomes of adverse pathology:

[1030] ● The gene combination of 1,831 / 2,047 (89.4%) had a statistically significant (LRS p value < 0.05) prognosis.

[1031] ● The gene combination of 1,679 / 2,047 (82.0%) increased the prognostic value (LRS p value <0.05) of the clinical information consisting of the best weighted biopsy information Gleason score, clinical stage and PSA.

[1032] ● 1,779 / 2,047 (86.9%) of the gene combinations increased the statistically significant (LRS p-value <0.05) prognostic value of clinical information composed of the Prostate Cancer Risk Assessment (CAPRA) score from 0 to 10 (Cooperberg MR, Pasta DJ, Elkin EP et al., The University of California, San Francisco Cancer of the Prostate Risk Assessment score: a straightforward and reliable preoperative predictor of disease recurrence after radical prostatectomy. Journal of Urology, 2005; 173:1938-42).

[1033] ● 1,798 / 2,047 (87.8%) gene combinations increased the statistically significant (LRS p-value <0.05) prognostic value for clinical information consisting of low, intermediate, or high risk categories from the European Association of Urology (EAU) (Mottet N, Bellmunt J, Briers E et al.: members of the EAU-ESTRO-ESUR-SIOG Prostate Cancer Guidelines Panel. EAU-ESTRO-ESUR-SIOG Guidelines on Prostate Cancer. Retrieved from: https: / / uroweb.org / guideline / prostate-cancer / #4).

[1034] For biochemical relapse as a secondary clinical outcome, the gene combination of 1,660 / 2,047 (81.1%) had a statistically significant (LRS p-value <0.05) prognosis.

[1035] Results of all 512 gene combinations including MTUS1 and TTC21B

[1036] A combination of 512 genes has a specific prognostic effect on aggressive prostate cancer. For pathology indicating poor primary clinical outcomes:

[1037] ● All 512 / 512 (100.0%) gene combinations had a statistically significant (LRS p-value < 0.05) prognosis (see [link to relevant documentation]). Figure 1 (A)).

[1038] ● All 512 / 512 (100.0%) gene combinations increased the prognostic value by a statistically significant (LRS p-value <0.05) percentage for clinical information consisting of the best-weighted biopsy information Gleason score, clinical stage, and PSA (see [link to relevant documentation]). Figure 1 (B)).

[1039] ● All 512 / 512 (100.0%) gene combinations showed a statistically significant (LRS p-value <0.05) increase in prognostic value based on clinical information from the Prostate Cancer Risk Assessment (CAPRA) score of 0 to 10 (see [link to relevant documentation]). Figure 1 (C)).

[1040] ● All 512 / 512 (100.0%) gene combinations increased the prognostic value (LRS p-value <0.05) of clinical information consisting of the European Association of Urology's low, intermediate, or high risk categories (see [link]). Figure 1 (D)).

[1041] For the secondary clinical outcome of biochemical relapse, the gene combination in 496 / 512 (96.9%) cases had a statistically significant (LRS p-value <0.05) prognosis (see [link to relevant documentation]). Figure 2 ).

[1042] Results of the 2-gene combination MTUS1 and TTC21B

[1043] For pathology of poor primary clinical outcomes, the 2-gene combination MTUS1 and TTC21B:

[1044] ● A statistically significant prognosis (LRS p-value = 0.0013) (see [link to relevant documentation]). Figure 1 (Molecular only, small image, red dot)

[1045] ●Increased the statistically significant (LRS p = 0.0101) prognostic value of clinical information composed of the best-weighted biopsy information Gleason score, clinical stage, and PSA (see [link to relevant documentation]). Figure 1 (Thumbnail for "Clinical Added Value", red dot)

[1046] ● The clinical information, comprised of the Prostate Cancer Risk Assessment (CAPRA) score from 0 to 10 (Ref A), significantly increased the prognostic value (LRS p = 0.0082) (see [link to relevant documentation]). Figure 1 (Capra Added Value thumbnail, red dot)

[1047] ● This clinical information, comprised of the European Association of Urology's low, intermediate, or high-risk categories (Ref B), was statistically significantly improved (LRS p = 0.0028) in terms of prognostic value (see [link]). Figure 1 (Thumbnail for "EAU Added Value", red dot)

[1048] For biochemical relapse as a secondary clinical outcome, the 2-gene combination MTUS1 and TTC21B had a statistically significant (LRS p = 0.0042) prognosis (see [link to relevant documentation]). Figure 2 (red dot).

[1049] Results of the 4-gene combination FOXM1, MCM3, MTUS1+ and TTC21B

[1050] For pathology of poor primary clinical outcomes, the 4-gene combination FOXM1, MCM3, MTUS1, and TTC21B:

[1051] ● A statistically significant prognosis (LRS p-value < 0.0001) (see [link to relevant documentation]). Figure 1 (A), Green Dot)

[1052] ●Increases the statistically significant (LRS p-value = 0.0001) prognostic value of clinical information composed of the best weighted biopsy information Gleason score, clinical stage, and PSA (see [link to relevant documentation]). Figure 1 (B), Green Dot)

[1053] ● The clinical information, comprised of a Prostate Cancer Risk Assessment (CAPRA) score from 0 to 10, significantly increased the prognostic value (LRS p-value = 0.0003) (see [link to relevant documentation]). Figure 1 (C), Green Dot)

[1054] ● This adds statistically significant (LRS p-value < 0.0001) prognostic value to clinical information comprised of the European Association of Urology's low, intermediate, or high risk categories (see [link]). Figure 1 (D), Green Dot)

[1055] For biochemical relapse as a secondary clinical outcome, the 4-gene combination FOXM1, MCM3, MTUS1, and TTC21B had a statistically significant (LRS p = 0.0009) prognosis (see [link to relevant documentation]). Figure 2 (Green dot).

[1056] discuss

[1057] In this study, genes identified as having individual prognostic properties for aggressive or recurrent prostate cancer were compared, with combinations of genes providing even stronger prognostic performance. Specifically, the combination of TTC21B and MTUS1, as a 2-gene prognostic marker, demonstrated superior prognostic performance.

[1058] The prognostic performance of the gene combination of the claimed invention provides a significant improvement over currently available individual clinicopathological variables. This improved prognostic performance enables more accurate risk stratification of patients diagnosed with cancers such as prostate cancer. On the one hand, accurate risk stratification can reduce unnecessary prescriptions for invasive treatments and associated side effects for patients with a lower risk of aggressive or recurrent cancer. On the other hand, identified patients with a higher risk of aggressive or recurrent cancer are correctly considered treatment candidates to reduce the risk of cancer invasiveness or recurrence. Therefore, accurate stratification provided by the method of the claimed invention has the potential to help reduce healthcare costs associated with controlling unnecessary side effects and costs associated with controlling aggressive or recurrent cancer due to inadequate treatment. Most importantly, accurate stratification generated by the method of the claimed invention can help improve the quality of life of patients who receive optimal treatment due to accurate stratification. Additionally, it has been reported that barriers to obtaining risk stratification information are the high costs associated with testing, partly due to molecular methods and large groups of biomarker analyses. The simplicity of the method of the claimed invention offers advantages over existing methods that employ larger prognostic marker groups by reducing assay costs and lowering technical barriers to introducing point-of-care, patient-friendly, and rapid prognostic testing.

[1059] In this specification, the terms “comprise,” “comprises,” “comprised,” and “comprising,” or any variations thereof, and the terms “include,” “includes,” “included,” and “including,” or any variations thereof, are considered to be completely interchangeable and should all be interpreted as broadly as possible, and vice versa.

[1060] The present invention is not limited to the embodiments described above, but may vary in both structure and details.

Claims

1. A method for predicting an individual's risk of invasive or recurrent prostate cancer, the method comprising the step of determining the expression of two genes, TTC21B and MTUS1, or proteins encoded by the genes, in a biological sample from the individual, wherein when the combination of the normalized expression values ​​of the two genes or the proteins encoded by the genes changes relative to the combination of the normalized expression values ​​of the two genes or the proteins encoded by the genes in a biological sample from a subject with non-invasive or non-recurrent prostate cancer, the individual is predicted to have an increased risk of invasive or recurrent prostate cancer.

2. A method for predicting the risk of prostate cancer recurrence or progression in an individual suffering from prostate cancer after treatment, the method comprising: The expression of two genes, TTC21B and MTUS1, or proteins encoded by said genes, in a biological sample obtained from said individual is determined, wherein when the combination of the normalized expression values ​​of said two genes or proteins encoded by said genes is changed relative to the combination of the normalized expression values ​​of said two genes in a biological sample obtained from a subject who did not have prostate cancer recurrence or progression after treatment, it is predicted that said individual has an increased risk of prostate cancer recurrence or a risk of prostate cancer progression after said treatment.

3. A method of determining a 5-year survival rate or a 10-year survival rate for an individual diagnosed with prostate cancer, the method comprising: The expression of two genes, TTC21B and MTUS1, or proteins encoded by these genes, is determined in a biological sample from the individual, wherein when the combination of the normalized expression values ​​of the two genes or proteins encoded by the genes is at least two genes from a biological sample obtained from a subject who survived prostate cancer 5 years or 10 years after diagnosis of prostate cancer, the individual is predicted to have a reduced 5-year survival rate or 10-year survival rate.

4. A method for identifying a prostate cancer patient suitable for treatment with a therapy for preventing recurrence or progression of prostate cancer, the method comprising: The expression of two genes, TTC21B and MTUS1, or proteins encoded by these genes, is determined in a biological sample from the individual, wherein when the combination of normalized expression values ​​of the genes or proteins encoded by the genes is the same or increased relative to the combination of normalized expression values ​​of the two genes in the biological sample from the same individual before administration of the therapy, the individual is predicted to be unsuitable for treatment with the therapy.

5. A method for monitoring the effectiveness of a prostate cancer treatment in an individual suffering from prostate cancer, the method comprising: The expression of two genes, TTC21B and MTUS1, or proteins encoded by these genes, is determined in a biological sample from the individual, wherein when the combination of normalized expression values ​​of the two genes or proteins encoded by the genes is the same or increased after administration of the treatment relative to the combination of normalized expression values ​​of the two genes in a biological sample from the same individual before administration of the treatment, the treatment is ineffective and the individual is predicted to have a poor outcome.

6. The method of claim 4 or claim 5, wherein the therapeutic therapy or treatment is active surveillance, neoadjuvant therapy, adjuvant therapy, surgery, or a combination thereof.

7. The method of claim 6, wherein the neoadjuvant therapy and the adjuvant therapy are agents selected from the group consisting of cyclophosphamide, methotrexate, 5-fluorouracil, gemcitabine, doxorubicin ( C 27 H 29 NO 11 ), paclitaxel ( C 47 H 51 NO 14 )、capecitabine ( C 15 H 22 FN3O6), docetaxel ( C 43 H 53 NO 14 )、cabazitaxel( C 45 H 57 NO 14 )、mitoxantrone ( C 22 H 28 N4O6) and estramustine (3-(bis(2-chloroethyl)carbamate) estradiol ester; C 23 H 31 Cl2NO3) and a luteinizing hormone-releasing (LHRH) agonist selected from leuprorelin ( C 59 H 84 N 16 O 12 )、goserelin ( C 59 H 84 N 18 O 14 )、triptorelin (triptorelin) C 64 H 82 N 18 O 13 ), euprolide mesylate (C 60 H 88 N 16 O 15 S), degarelix ( C 82 H 103 C1N 18 O 16 ) and relugolix ( C 29 H 27 F2N7O5S).

8. according to the method described in any one of the preceding claims, it further comprises the step of measuring the expression of at least one gene or the protein encoded by the gene in the biological sample from the individuality, the gene being selected from MCM3, FOXM1, CUL4A, GPC1, PCNA, IRF7, BRIP1, ULK1 and SPAG5.

9. The method of claim 8, wherein the at least one gene is MCM3.

10. The method according to any one of claims 1 to 7, further comprising the step of determining the expression of at least two genes in the biological sample from the individual, wherein the at least two genes are selected from CUL4A, MCM3, GPC1, PCNA, IRF7, BRIP1, ULK1, SPAG5 and FOXM1. The method according to claim 10 , wherein the at least two genes are MCM3 and FOXM1.

12. A method for predicting an individual's risk of developing an aggressive or recurrent cancer, the method comprising the step of determining the expression of two genes, TTC21B and MTUS1, or proteins encoded by the genes, in a biological sample from the individual, wherein when the combination of normalized expression values ​​of the two genes or proteins encoded by the genes is changed relative to the combination of normalized expression values ​​of the two genes or proteins encoded by the genes in a biological sample from a subject with non-aggressive or non-recurrent cancer, the individual is predicted to have an increased risk of developing an aggressive or recurrent cancer.

13. A method for predicting the risk of cancer recurrence or progression in an individual suffering from cancer after treatment, the method comprising: The expression of two genes, TTC21B and MTUS1, or proteins encoded by the genes, is determined in a biological sample obtained from the individual, wherein when the combination of the normalized expression values ​​of the two genes or the proteins encoded by the genes is changed relative to the combination of the normalized expression values ​​of the two genes from a biological sample obtained from a subject who did not have cancer recurrence or progression after treatment, the individual is predicted to have an increased risk of cancer recurrence or cancer progression after the treatment.

14. A method of determining a 5-year survival rate or a 10-year survival rate for an individual diagnosed with cancer, the method comprising: The expression of two genes, TTC21B and MTUS1, or proteins encoded by these genes, is determined in a biological sample from the individual, wherein when the combination of normalized expression values ​​of the two genes or proteins encoded by the genes is changed relative to the combination of normalized expression values ​​of the two genes from a biological sample obtained from a subject who survived cancer 5 or 10 years after cancer diagnosis, the individual is predicted to have a decreased 5-year survival rate or 10-year survival rate.

15. A method of identifying a cancer patient suitable for treatment with a therapy for preventing cancer recurrence or progression, the method comprising: The expression of two genes, TTC21B and MTUS1, or proteins encoded by these genes, in a biological sample from the individual is determined, wherein when the combination of normalized expression values ​​of the two genes or proteins encoded by the genes is the same or increased relative to the combination of normalized expression values ​​of the two genes in a biological sample from the same individual before administration of the therapy, the individual is predicted to be unsuitable for treatment with the therapy.

16. A method for monitoring the effectiveness of a cancer treatment in an individual suffering from cancer, the method comprising: The expression of two genes, TTC21B and MTUS1, or proteins encoded by these genes, is determined in a biological sample from the individual, wherein when the combination of normalized expression values ​​of the two genes is the same or increased after administration of the treatment relative to the combination of normalized expression values ​​of the two genes, or proteins encoded by the genes, in a biological sample from the same individual before administration of the treatment, the treatment is ineffective and the individual is predicted to have a poor outcome.

17. The method of any one of claims 15 or 16, wherein the therapeutic therapy is active surveillance, neoadjuvant therapy, adjuvant therapy, surgery, or a combination thereof.

18. The method of claim 17, wherein the neoadjuvant therapy and the adjuvant therapy are agents selected from the group consisting of cyclophosphamide, methotrexate, 5-fluorouracil, gemcitabine, doxorubicin ( C 27 H 29 NO 11 ), paclitaxel ( C 47 H 51 NO 14 ), Capecitabine ( C 15 H 22 FN3O6), docetaxel ( C 43 H53 NO 14 )、Cabataxel( C 45 H 57 NO 14 )、mitoxantrone( C 22 H 28 N4O6) and estramustine (3-(bis(2-chloroethyl)carbamate) estradiol ester; C 23 H 31 Cl2NO3) and a luteinizing hormone-releasing (LHRH) agonist, wherein the LHRH agonist is selected from leuprolide ( C 59 H 84 N 16 O 12 )、Goserelin( C 59 H 84 N 18 O 14 )、triptorelin( C 64 H 82 N 18 O 13 ), leuprolide mesylate (C 60 H 88 N 16 O 15 S), Degarelix C 82 H 103 C1N 18 O 16 ) and Ruilugoli C 29 H 27 F2N7O5S).

19. The method according to any one of claims 12 to 18, further comprising the step of determining the expression of at least one gene or a protein encoded by the gene in the biological sample from the individual, the gene being selected from the group consisting of MCM3, FOXM1, CUL4A, GPC1, PCNA, IRF7, BRIP1, ULK1 and SPAG5.

20. The method of claim 19, wherein the at least one gene is MCM3.

21. The method according to any one of claims 16 to 18, further comprising the step of determining the expression of at least two genes in the biological sample from the individual, the at least two genes being selected from CUL4A, MCM3, GPC1, PCNA, IRF7, BRIP1, ULK1, SPAG5 and FOXM1.

22. The method of claim 21, wherein the at least two genes are MCM3 and FOXM1.

23. The method according to any one of claims 12 to 22, wherein the cancer is selected from the group comprising: multiple myeloma, prostate cancer, glioblastoma, lymphoma, fibrosarcoma; myxosarcoma; liposarcoma; chondrosarcoma; osteogenic sarcoma; chordoma; angiosarcoma; endotheliosarcoma; lymphangiosarcoma; lymphangioendotheliosarcoma, synovialoma; mesothelioma; Ewing's tumor (Ewing's tumour, leiomyosarcoma; rhabdomyosarcoma; colon cancer; pancreatic cancer; ovarian cancer; squamous cell carcinoma; basal cell carcinoma; adenocarcinoma; sweat gland carcinoma; sebaceous gland carcinoma; papillary carcinoma; papillary adenocarcinoma; cystadenocarcinoma; medullary carcinoma; bronchogenic carcinoma; renal cell carcinoma; hepatoma; bile duct carcinoma; choriocarcinoma; seminoma, embryonal carcinoma; Wilms' tumour; cervical cancer; uterine cancer; testicular tumors; lung cancer; small cell lung cancer; bladder cancer; epithelial carcinoma; glioma; astrocytoma; medulloblastoma; craniopharyngioma; ependymoma; pinealoma; hemangioblastoma; acoustic neuroma; oligodendroglioma; meningioma; melanoma; retinoblastoma; head and neck cancer types; bladder cancer types; colorectal cancer; and leukemia.

24. The method of claim 23, wherein the cancer is prostate cancer.

25. The method of any one of claims 12 to 22, wherein the cancer is a precancer selected from the group comprising Barrett's oesophagus, Bowen's disease, familial adenomatous polyposis, lobular carcinoma in situ, Lynch syndrome, MEN2, vaginal intraepithelial neoplasia, and vulvar intraepithelial neoplasia.

26. A method for predicting the risk of an invasive or recurrent cancer in an individual, the method comprising the step of determining the expression of at least two genes or proteins encoded by the genes in a biological sample from the individual, the genes being selected from the group consisting of TTC21B, MCM3, MTUS1, FOXM1, CUL4A, GPC1, PCNA, IRF7, BRIP1, ULK1 and SPAG5, wherein when the combination of the normalized expression values ​​of the at least two genes or proteins encoded by the genes is altered relative to the combination of the normalized expression values ​​of the at least two genes or proteins encoded by the genes in a biological sample from a subject with non-invasive or non-recurrent cancer, the individual is predicted to have an increased risk of having an invasive or recurrent cancer.

27. A method for predicting the risk of cancer recurrence or progression in an individual suffering from cancer after treatment, the method comprising: Determining the expression of at least two genes, or proteins encoded by the genes, in a biological sample obtained from the individual, wherein the genes are selected from TTC21B, MCM3, MTUS1, FOXM1, CUL4A, GPC1, PCNA, IRF7, BRIP1, ULK1 and SPAG5, wherein when the combination of the normalized expression values ​​of the at least two genes, or proteins encoded by the genes, is changed relative to the combination of the normalized expression values ​​of the at least two genes in a biological sample obtained from a subject who did not have cancer recurrence or progression after treatment, the individual is predicted to have an increased risk of cancer recurrence or cancer progression after the treatment.

28. A method of determining a 5-year survival rate or a 10-year survival rate for an individual diagnosed with cancer, the method comprising: The expression of at least two genes or proteins encoded by these genes in a biological sample from the individual is determined, and the genes are selected from TTC21B, MCM3, MTUS1, FOXM1, CUL4A, GPC1, PCNA, IRF7, BRIP1, ULK1 and SPAG5, wherein when the combination of normalized expression values ​​of the at least two genes or proteins encoded by the genes is changed relative to the combination of normalized expression values ​​of the at least two genes from a biological sample obtained from a subject who survived cancer 5 years or 10 years after cancer diagnosis, it is predicted that the individual has a decreased 5-year survival rate or 10-year survival rate.

29. A method of identifying a cancer patient suitable for treatment with a therapy for preventing cancer recurrence or progression, the method comprising: Determine the expression of at least two genes or proteins encoded by these genes in a biological sample from the individual, wherein the genes are selected from TTC21B, MCM3, MTUS1, FOXM1, CUL4A, GPC1, PCNA, IRF7, BRIP1, ULK1 and SPAG5, wherein when the combination of normalized expression values ​​of the at least two genes or proteins encoded by the genes is the same or increased after the administration of the therapy relative to the combination of normalized expression values ​​of the at least two genes in the biological sample from the same individual before the administration of the therapy, it is predicted that the individual is not suitable for treatment with the therapy.

30. A method for monitoring the effectiveness of a cancer treatment in an individual suffering from prostate cancer, the method comprising: The expression of at least two genes or proteins encoded by these genes in a biological sample from the individual is determined, and the genes are selected from TTC21B, MCM3, MTUS1, FOXM1, CUL4A, GPC1, PCNA, IRF7, BRIP1, ULK1 and SPAG5, wherein when the combination of normalized expression values ​​of the at least two genes or proteins encoded by the genes is the same or increased after administration of the treatment relative to the combination of normalized expression values ​​of the at least two genes in the biological sample from the same individual before administration of the treatment, the treatment is ineffective and the individual is predicted to have a poor outcome.

31. The method of claim 29 or 30, wherein the therapeutic therapy or treatment is active surveillance, neoadjuvant therapy, adjuvant therapy, surgery, or a combination thereof.

32. The method of claim 31, wherein the neoadjuvant therapy and the adjuvant therapy are agents selected from the group consisting of cyclophosphamide, methotrexate, 5-fluorouracil, gemcitabine, doxorubicin ( C 27 H 29 NO 11 ), paclitaxel ( C 47 H 51 NO 14 ), Capecitabine ( C 15 H 22 FN3O6), docetaxel ( C 43 H 53 NO 14 )、Cabataxel( C 45 H 57 NO 14 )、mitoxantrone( C 22 H 28 N4O6) and estramustine (3-(bis(2-chloroethyl)carbamate) estradiol ester; C 23 H 31 Cl2NO3) and a luteinizing hormone-releasing (LHRH) agonist, wherein the LHRH agonist is selected from leuprolide ( C 59 H 84 N 16 O 12 )、Goserelin( C 59 H 84 N 18 O 14)、triptorelin( C 64 H 82 N 18 O 13 ), leuprolide mesylate (C 60 H 88 N 16 O 15 S), Degarelix C 82 H 103 C1N 18 O 16 ) and Ruilugoli ( C 29 H 27 F2N7O5S).

33. according to the method described in any one of claims 26 to 32, wherein the first gene of the at least two genes is selected from TTC21B, PCNA, IRF7, ULK1, SPAG5 and FOXM1, and the second gene of the at least two genes is selected from BRIP1, CUL4A, GPC1, MCM3 and MTUS1; or wherein the at least two genes are optionally selected from TTC21B, PCNA, IRF7, ULK1, SPAG5 and FOXM1; or wherein the at least two genes are optionally selected from BRIP1, CUL4A, GPC1, MCM3 and MTUS1.

34. The method according to any one of claims 26 to 32, wherein the at least two genes selected are MTUS1 and MCM3.

35. The method according to any one of claims 26 to 32, wherein the at least two genes selected are TTC21B and MCM3.

36. The method according to any one of claims 26 to 32, wherein the at least two genes selected are TTC21B and MTUS1.

37. The method of any one of claims 26 to 32, wherein at least three genes are selected from CUL4A, TTC21B, MCM3, GPC1, PCNA, MTUS1, IRF7, BRIP1, ULK1, SPAG5, and FOXM1, and wherein the three genes are TTC21B, MCM3, and MTUS1.

38. The method of any one of claims 26 to 32, wherein at least four genes are selected from CUL4A, TTC21B, MCM3, GPC1, PCNA, MTUS1, IRF7, BRIP1, ULK1, SPAG5, and FOXM1, and wherein the four genes are TTC21B, MCM3, MTUS1, and FOXM1.

39. The method of any one of claims 26 to 38, wherein the cancer is selected from the group consisting of: multiple myeloma, prostate cancer, glioblastoma, lymphoma, fibrosarcoma; myxosarcoma; liposarcoma; chondrosarcoma; osteogenic sarcoma; chordoma; angiosarcoma; endotheliosarcoma; lymphangiosarcoma; lymphangioendotheliosarcoma, synovialoma; mesothelioma; Ewing's tumor tumour, leiomyosarcoma; rhabdomyosarcoma; colon cancer; pancreatic cancer; ovarian cancer; squamous cell carcinoma; basal cell carcinoma; adenocarcinoma; sweat gland carcinoma; sebaceous gland carcinoma; papillary carcinoma; papillary adenocarcinoma; cystadenocarcinoma; medullary carcinoma; bronchogenic carcinoma; renal cell carcinoma; hepatoma; bile duct carcinoma; choriocarcinoma; seminoma, embryonal carcinoma; Wilms' tumour; cervical cancer; uterine cancer; testicular tumors; lung cancer; small cell lung cancer; bladder cancer; epithelial carcinoma; glioma; astrocytoma; medulloblastoma; craniopharyngioma; ependymoma; pinealoma; hemangioblastoma; acoustic neuroma; oligodendroglioma; meningioma; melanoma; retinoblastoma; head and neck cancer types; bladder cancer types; colorectal cancer; and leukemia.

40. The method of claim 39, wherein the cancer is prostate cancer.

41. The method of any one of claims 26 to 38, wherein the cancer is a precancer selected from the group comprising Barrett's esophagus, Bowen's disease, familial adenomatous polyposis, lobular carcinoma in situ, Lynch syndrome, MEN2, vaginal intraepithelial neoplasia, and vulvar intraepithelial neoplasia.

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