Gene expression profiling for prognosis in adrenocortical carcinoma

Abstract

The invention discloses a group of genes for adrenocortical carcinoma prognosis assessment, belonging to the field of biological and medical testing. The genes involved include: ASPM, C3, CCNB2, CDCA3, CDCA5, CENPM, CENPW, CEP95 (CCDC45), DNA2, DNAJC9, ECT2, EME1, FANCI, G0S2, HAUS8, HELLS, H1F0, KIF11, LMO7, MCM10, MXD3, NUP188, NVL, PKMYT1, PSRC1, TACC3, TOMM5, TRIP13, UBE2T and other 29 genes. It involves detecting the expression level of the products (RNA, protein) of the above genes (single or any combination) to perform molecular typing and prognosis assessment on adrenocortical carcinoma; a reliable method is provided for rapidly predicting the molecular typing and prognosis of adrenocortical carcinoma.

Description

Technical Field

[0001] The present invention belongs to the field of biological and medical testing, and is a method and application for determining the malignancy and prognosis of adrenocortical carcinoma by detecting changes in the expression of a single or any combination of related gene products (protein, RNA) in biological samples such as adrenocortical carcinoma and body fluids. The genes involved include: ASPM, C3, CCNB2, CDCA3, CDCA5, CENPM, CENPW, CEP95 (CCDC45), DNA2, DNAJC9, ECT2, EME1, FANCI, G0S2, HAUS8, HELLS, H1F0, KIF11, LMO7, MCM10, MXD3, NUP188, NVL, PKMYT1, PSRC1, TACC3, TOMM5, TRIP13, UBE2T and other 29 genes. Background Art

[0002] Adrenocortical carcinoma (ACC) is a kind of morbidity is low, but has invasive tumor, and some patients have poor prognosis. Rapid identification of the malignancy of adrenocortical carcinoma, molecular typing of prognosis, is of great value to the selection of treatment plan, etc. In the present invention, we find a group of genes related to the prognosis of adrenocortical carcinoma by large-scale screening of the relationship between adrenocortical carcinoma gene expression profile and prognosis: ASPM, C3, CCNB2, CDCA3, CDCA5, CENPM, CENPW, CEP95 (CCDC45), DNA2, DNAJC9, ECT2, EME1, FANCI, G0S2, HAUS8, HELLS, H1F0, KIF11, LMO7, MCM10, MXD3, NUP188, NVL, PKMYT1, PSRC1, TACC3, TOMM5, TRIP13, 29 genes such as UBE2T. The combined application of the above genes is beneficial to the prognosis determination and molecular typing of adrenocortical carcinoma.

[0003] Based on the research foundation and current status of the prior art, the inventor of the present application intends to provide an indicator for the prognosis and molecular typing of adrenocortical carcinoma. The present invention relates to predicting the malignancy and prognosis of adrenocortical carcinoma by the expression level of the above-mentioned gene set (single or any combination of two or more) in adrenocortical carcinoma tissue. In particular, by combining multiple genes to form an array, and then combining it with the extraction and detection of proteins (such as immunoturbidimetry, colloidal gold, immunochemiluminescence, etc.) or RNA (such as RT-PCR, fluorescent quantitative PCR, RNA array, RNA Sequencing, etc.) in adrenocortical carcinoma tissue, a rapid, high-throughput, effective and objective method can be provided for the molecular typing and prognosis evaluation of adrenocortical carcinoma. Summary of the invention

[0004] The purpose of the present invention is to provide a detection index for "rapidly and efficiently evaluating the molecular typing and prognosis of adrenocortical carcinoma". A method for identifying the molecular typing and prognosis of adrenocortical carcinoma by detecting the expression spectrum of related genes.

[0005] The present invention screens out a group of genes highly correlated with the prognosis of adrenocortical carcinoma based on the analysis of large-scale gene expression profiles, including 29 genes such as ASPM, C3, CCNB2, CDCA3, CDCA5, CENPM, CENPW, CEP95, DNA2, DNAJC9, ECT2, EME1, FANCI, G0S2, HAUS8, HELLS, H1F0, KIF11, LMO7, MCM10, MXD3, NUP188, NVL, PKMYT1, PSRC1, TACC3, TOMM5, TRIP13, and UBE2T. The molecular typing and prognosis evaluation of adrenocortical carcinoma are related to the expression of the above genes (single or in any combination) in adrenocortical carcinoma, and can be used to predict the molecular typing, drug resistance, drug screening, and patient prognosis analysis of adrenocortical carcinoma;

[0006] S1 uses the above gene expression levels alone to determine the molecular typing of adrenocortical carcinoma and the prognosis of patients;

[0007] S2 combines the above genes (any combination of 2 or more) with expression changes to determine the molecular typing of adrenocortical carcinoma and the prognosis of patients;

[0008] S3 The above single gene or any combination is combined with other indicators to determine the molecular typing of adrenocortical carcinoma and the prognosis of patients;

[0009] The present invention relates to the detection of the above gene expression products including: protein and RNA.

[0010] The present invention relates to the above-mentioned gene products to be detected, including those in tumor cells, as well as mRNA, proteins and exosomes that enter body fluids for various reasons, including complete and fragmentary ones.

[0011] The detection objects of the present invention include: tumor tissue, body fluids (including blood, urine, puncture fluid, etc.), puncture tissue, etc., as molecular typing and patient evaluation indicators for predicting adrenocortical carcinoma;

[0012] The body fluids involved in the present invention include blood, urine, tumor puncture fluid, lymph fluid, etc.

[0013] The blood involved in the present invention includes whole blood, serum, plasma, separated nucleated cells, circulating tumor cells in the blood, exosomes, etc.

[0014] The above gene protein detection involved in the present invention includes directly extracting adrenocortical carcinoma tissue protein and performing detection and immunohistochemical analysis by chemical or immunological methods.

[0015] The above gene RNA detection involved in the present invention includes directly extracting RNA from adrenocortical carcinoma tissue and then combining it with RT-PCR technology, fluorescent quantitative PCR technology, RNA Sequencing, RNA array, etc. for detection.

[0016] The present invention relates to the detection of gene products including protein level and RNA level, and the detection methods involve: immunohistochemistry, immunofluorescence, Western blot, ELISA, flow cytometry, RT-PCR, colloidal gold, immunological detection, chemical detection, fluorescent quantitative PCR, RNA Sequencing, RNA array and other detection technologies (but not limited to these technologies).

[0017] The RT-PCR involved in the present invention detects the RNA product of the above gene. After reverse transcription, the PCR involved includes: ordinary PCR, fluorescent quantitative PCR, nested PCR, multiplex PCR, digital PCR, etc.

[0018] The beneficial effects of the present invention are:

[0019] Micro, rapid, and high-throughput are the general trend of technological development; although pathological morphology is the gold standard for diagnosing tumors, it has limited effect on tumor prognosis assessment. Therefore, molecular biological indicators are expected to become an important means of tumor prognosis assessment and molecular typing. In the present invention, we provide a molecular method for molecular typing, drug screening, prognosis assessment, etc. of adrenocortical carcinoma based on changes in tumor genome expression profiles. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 .The heat map of mRNA Sequencing results shows: the overall expression of mRNA of the genes ASPM, C3, CCNB2, CDCA3, CDCA5, CEP95 (CCDC45), CENPM, CENPW, DNA2, DNAJC9, ECT2, EME1, FANCI, G0S2, HAUS8, HELLS, etc. involved in the present invention in adrenal cortical carcinoma (ACC) tissue.

[0021] Figure 2.The heat map of mRNA Sequencing results shows: the overall expression of mRNA of the genes KRT5, KRT6A, KRT7, KRT16, KRT6B, KRT15, ​​S100A2, S100A14, S100A9, S100A8, SFN, SERPINB3, LAD1, FXYD3, TACSTD2, PI3, CXCL17, MAL2, CSTA, PDZK1IP1, LAMB3, RAB25, C19orf33, ELF3, etc. involved in the present invention in adrenal cortical carcinoma (ACC) tissue.

[0022] Figure 3 The survival curve results showed the relationship between the mRNA expression levels of ASPM, C3, CCNB2, CDCA3, CDCA5, CEP95 (CCDC45), CENPM, CENPW, DNA2, DNAJC9, ECT2, EME1, FANCI, G0S2, HAUS8, HELLS and the overall survival time of patients with adrenocortical carcinoma (ACC). Among them, patients with high expression levels of ASPM, CCNB2, CDCA3, CDCA5, CEP95, CENPM, CENPW, DNA2, DNAJC9, ECT2, EME1, FANCI, HAUS8, HELLS had a higher tumor malignancy and a worse prognosis; while patients with high expression levels of C3, G0S2 and other genes had a lower tumor malignancy and a better prognosis.

[0023] Figure 4 The survival curve results showed the relationship between the mRNA expression levels of genes such as KIF11, LMO7, MCM10, MXD3, NUP188, NVL, PKMYT1, PSRC1, TACC3, TOMM5, TRIP13, UBE2T, and H1F0 and the overall survival time of patients with adrenocortical carcinoma (ACC). Patients with high expression levels generally had a higher degree of tumor malignancy and a worse prognosis. DETAILED DESCRIPTION

[0024] Example 1

[0025] In the present invention, we take adrenocortical carcinoma (ACC) as the implementation object, and use RNA Sequencing technology to analyze the relationship between mRNA expression in adrenocortical carcinoma tissue and patient prognosis as an example. Based on the relationship between the mRNA expression of adrenocortical carcinoma tissue genes and prognosis in the study of The Cancer Genome Atlas (TCGA) database, we screened a group of genes closely related to patient prognosis, including: ASPM, C3, CCNB2, CDCA3, CDCA5, CENPM, CENPW, CEP95 (CCDC45), DNA2, DNAJC9, ECT2, EME1, FANCI, G0S2, HAUS8, HELLS, H1F0, KIF11, LMO7, MCM10, MXD3, NUP188, NVL, PKMYT1, PSRC1, TACC3, TOMM5, TRIP13, UBE2T and other 29 genes ( Figure 1 , 2).

[0026] Through the study of the relationship between the mRNA Sequencing results and the overall survival curve of patients in the TCGA database, we found that the expression levels of ASPM, CCNB2, CDCA3, CDCA5, CENPM, CENPW, CEP95 (CCDC45), DNA2, DNAJC9, ECT2, EME1, FANCI, HAUS8, HELLS, H1F0, KIF11, LMO7, MCM10, MXD3, NUP188, NVL, PKMYT1, PSRC1, TACC3, TOMM5, TRIP13, UBE2T and other genes indicated that adrenocortical carcinoma had a higher degree of malignancy and a poorer prognosis; while the higher expression levels of C3, G0S2 and other genes indicated that adrenocortical carcinoma had a lower degree of malignancy and a better prognosis ( Figure 3 , 4).

[0027] In the present invention, we show that the mRNA Sequencing results in the TCGA database show that there are also differences in the above gene expression profiles. Adrenocortical carcinoma can be molecularly classified based on the differences in the above gene expression profiles; for example: ASPM high / CCNB2 high / CDCA3 high / CDCA5 high The type is more malignant and has a poor prognosis; ANXA8 low / LGALS7B low / CALML3 low / PKP1 lowAdrenocortical carcinoma is divided into different molecular subtypes ( Figure 1 , 2).

[0028] summary

[0029] Our results show that 29 genes including ASPM, C3, CCNB2, CDCA3, CDCA5, CENPM, CENPW, CEP95 (CCDC45), DNA2, DNAJC9, ECT2, EME1, FANCI, G0S2, HAUS8, HELLS, H1F0, KIF11, LMO7, MCM10, MXD3, NUP188, NVL, PKMYT1, PSRC1, TACC3, TOMM5, TRIP13, and UBE2T are closely related to adrenocortical carcinoma, and the expression levels of the above genomic mRNA can be used for molecular typing and prognostic evaluation of adrenocortical carcinoma. Tip: The present invention provides reliable markers for molecular typing and prognostic evaluation of adrenocortical carcinoma.

Claims

1. Use of a gene detection reagent in the preparation of a prognosis assessment product for adrenocortical carcinoma, characterized in that: The gene is CDCA3; The application is carried out by detecting the expression level of the gene in adrenocortical carcinoma tissue or body fluid of adrenocortical carcinoma patients; Among them, high CDCA3 expression indicates high malignancy of adrenocortical carcinoma and poor prognosis of patients.

2. The use according to claim 1, characterized in that: The adrenocortical carcinoma tissue includes tumor tissue and puncture tissue; the body fluid of the adrenocortical carcinoma patient includes blood, urine and puncture fluid.

3. The use according to claim 2, characterized in that: The blood includes whole blood, serum and plasma.

4. The use according to claim 1, characterized in that: The gene expression products include protein expression products and RNA expression products.

5. The use according to claim 4, characterized in that: The method for detecting the protein expression level includes directly extracting adrenocortical carcinoma tissue protein and performing detection and immunohistochemical analysis by chemical or immunological methods. The method for detecting the RNA expression level includes directly extracting adrenocortical carcinoma tissue RNA for detection.

6. The use according to claim 5, characterized in that: The gene expression detection method includes RT-PCR technology, immunological detection technology, fluorescent quantitative PCR technology, RNA Sequencing technology and RNAarray technology.

7. The use according to claim 6, characterized in that: When the RT-PCR technique is used to detect the gene RNA expression level, after reverse transcription, the PCR techniques used include: conventional PCR, fluorescent quantitative PCR, nested PCR, multiplex PCR, digital PCR and chip microarray.

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