Application of FKBP9 detection agent in preparation of products for liver cancer diagnosis or prognosis evaluation

Abstract

The invention discloses application of an FKBP9 detection agent in preparation of products for liver cancer diagnosis, auxiliary diagnosis, screening, auxiliary screening, prognosis evaluation, risk assessment and / or risk stratification. The content of the FKBP9 in blood of a healthy control person and a patient with advanced liver cancer is remarkably different, and the FKBP9 has high sensitivity and specificity and can be used as a biomarker for diagnosing liver cancer. The content of the serum FKBP9 in a healthy control person and a patient with the advanced liver cancer is detected by adopting an enzyme-linked immunosorbent assay, and after a large number of clinical samples are subjected to immunoassay, it is determined that the content of the serum FKBP9 in the healthy control person and the content of the serum FKBP9 in the patient with the advanced liver cancer have significant difference; and ROC curve analysis is adopted to further verify that the serum FKBP9 can be used as a serum biomarker for diagnosing liver cancer. Meanwhile, it is also verified that FKBP9 in blood can be used as a biomarker for prognosis evaluation of liver cancer.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and particularly relates to the use of an FKBP9 detection agent in the preparation of products for liver cancer diagnosis or prognosis evaluation. Background Art

[0002] Liver cancer is one of the top three cancers in terms of mortality, and its incidence ranks fifth in the global cancer incidence. Liver cancer is divided into primary liver cancer and secondary liver cancer, and primary liver cancer is the most common liver cancer. Among them, the vast majority of primary liver cancers are hepatocellular carcinoma (HCC). The onset of HCC is affected by both environmental and dietary factors. Viruses, mycin, and alcohol can all induce hepatocellular carcinoma. In addition, due to the easy metastasis tendency of liver cancer and adverse reactions to drug treatment, the prognosis after treatment is also poor.

[0003] With the development of molecular biology research, more and more studies have found that molecular biomarkers are closely related to the occurrence, development and prognosis of liver cancer. Liver cancer cells have different biological activities from normal cells and can secrete different marker antigens into the patient's blood. The level of tumor marker antigens in normal human blood is very low. Therefore, by detecting the level of tumor markers in the blood, it can provide clues for the diagnosis or screening of primary liver cancer.

[0004] However, the sensitivity and specificity of existing biomarkers cannot meet the needs of clinical diagnosis of liver cancer. Further screening of efficient and sensitive biomarkers is one of the most effective methods to achieve liver cancer screening, diagnosis and prognosis. Summary of the invention

[0005] The purpose of the present invention is to provide an application of a FKBP9 detection agent in the preparation of a product for liver cancer diagnosis or prognosis evaluation, so as to solve the problem that the sensitivity and specificity of existing biomarkers cannot meet the needs of clinical diagnosis of liver cancer.

[0006] To achieve the above object, the present invention adopts the following technical solution: In a first aspect, the present invention provides a biomarker for diagnosis or prognosis assessment of liver cancer, wherein the biomarker is FKBP9.

[0007] In a second aspect, the present invention provides a kit for liver cancer diagnosis or prognosis assessment, the kit comprising a FKBP9 detection agent for detecting FKBP9 expression.

[0008] In a third aspect, the present invention provides the use of a FKBP9 detection agent in the preparation of products for liver cancer diagnosis, auxiliary diagnosis, screening, auxiliary screening, prognosis evaluation, risk assessment and / or risk stratification.

[0009] In some embodiments, the liver cancer diagnosis, auxiliary diagnosis, screening and / or auxiliary screening comprises the following steps: (A1) determining the expression level of FKBP9 in a blood sample from a subject; (A2) Comparison of FKBP9 expression levels with baseline values, Wherein an increased FKBP9 expression level compared to a baseline value indicates that the subject has liver cancer or is at risk of developing liver cancer.

[0010] In some embodiments, the blood sample is from a subject suspected of having liver cancer. In some embodiments, the blood sample is from a subject suspected of being at risk of developing liver cancer. Risk factors associated with liver cancer include, but are not limited to, hepatitis B virus and / or hepatitis C virus infection, excessive drinking, liver disease associated with hepatic steatosis or metabolic dysfunction, exposure to aflatoxin B1 in the diet, cirrhosis caused by various other causes, and a family history of liver cancer.

[0011] In some embodiments, the prognostic assessment comprises the following steps: (B1) Determining the expression level of FKBP9 in blood samples from liver cancer patients; (B2) Comparison of FKBP9 expression levels with baseline values, The increased FKBP9 expression level compared with the baseline value indicates that the liver cancer patient has a poor prognosis.

[0012] In some embodiments, the blood sample is from a patient with liver cancer who is currently receiving treatment. In some embodiments, the blood sample is from a patient with liver cancer who has not yet received treatment. Liver cancer treatments include, but are not limited to, liver resection, liver transplantation, ablation therapy, intravascular interventional therapy, radiotherapy, systemic anti-tumor therapy, traditional Chinese medicine treatment, etc. Systemic anti-tumor therapy includes but is not limited to molecular targeted drug therapy, immune checkpoint inhibitor therapy, chemotherapy and traditional Chinese medicine treatment, such as atezolizumab, bevacizumab, durvalumab, carrelizumab, ramucirumab, nivolumab, pembrolizumab, tilelizumab, tesimumab, sintilimab, apatinib, ipilimumab, donafenib, apatinib mesylate, cabozantinib, lenvatinib, regorafenib, sorafenib or their combinations, such as FOLFOX4 regimen and STRIDE regimen; in addition, it also includes treatment for the underlying diseases of liver cancer, such as antiviral therapy, liver protection and choleretic treatment, and supportive symptomatic treatment.

[0013] In some embodiments, the prognostic assessment indicator is selected from one or more of overall survival, disease-specific survival, progression-free survival, and risk of liver cancer cell metastasis.

[0014] In other aspects, the present invention provides a method (eg, an in vitro method) for diagnosing, assisting in diagnosis, screening and / or assisting in screening of liver cancer, which is based on the expression level of FKBP9 in a blood sample from a subject.

[0015] In some embodiments, the method for diagnosing, assisting diagnosis, screening and / or assisting screening of liver cancer comprises: a) obtaining a blood sample from the subject; b) determining the expression level of FKBP9 in the blood sample, wherein an elevated FKBP9 expression level compared to a baseline value indicates that the subject suffers from liver cancer or is at risk of suffering from liver cancer. In some embodiments, the blood sample is from a subject suspected of suffering from liver cancer. In some embodiments, the blood sample is from a subject suspected of being at risk of suffering from liver cancer. Risk factors associated with liver cancer include, but are not limited to, hepatitis B virus and / or hepatitis C virus infection, excessive drinking, liver disease associated with hepatic steatosis or metabolic dysfunction, exposure to aflatoxin B1 in the diet, cirrhosis caused by various other causes, and a family history of liver cancer.

[0016] In other aspects, the present invention provides a method (eg, an in vitro method) for liver cancer prognosis assessment based on the FKBP9 expression level in a blood sample from a liver cancer patient.

[0017] In some embodiments, the method for evaluating the prognosis of liver cancer comprises: a) obtaining a blood sample from the liver cancer patient; b) determining the expression level of FKBP9 in the blood sample, wherein an increased expression level of FKBP9 compared to a baseline value indicates that the liver cancer patient has a poor prognosis. In some embodiments, the blood sample is from a liver cancer patient who is undergoing treatment. In some embodiments, the blood sample is from a liver cancer patient who has not yet been treated. Liver cancer treatment includes but is not limited to liver resection, liver transplantation, ablation therapy, intravascular interventional therapy, radiotherapy, systemic anti-tumor therapy, traditional Chinese medicine treatment, etc. Systemic anti-tumor therapy includes but is not limited to molecular targeted drug therapy, immune checkpoint inhibitor therapy, chemotherapy and traditional Chinese medicine treatment, such as atezolizumab, bevacizumab, durvalumab, carrelizumab, ramucirumab, nivolumab, pembrolizumab, tilelizumab, tesimumab, sintilimab, apatinib, ipilimumab, donafenib, apatinib mesylate, cabozantinib, lenvatinib, regorafenib, sorafenib or their combinations, such as FOLFOX4 regimen and STRIDE regimen; in addition, it also includes treatment for the underlying diseases of liver cancer, such as antiviral therapy, liver protection and choleretic treatment, and supportive symptomatic treatment.

[0018] In other aspects, the present invention provides a product for diagnosis, auxiliary diagnosis, screening and / or auxiliary screening and / or prognostic evaluation of liver cancer, which comprises one or more FKBP9 detection agents.

[0019] In some embodiments, the product for liver cancer diagnosis, auxiliary diagnosis, screening and / or auxiliary screening and / or prognostic evaluation comprises one or more FKBP9 detection agents, which are used to determine the FKBP9 expression level in a blood sample from a subject or a liver cancer patient. In some embodiments, the product further comprises an operation guide for using a FKBP9 detection agent to determine the FKBP9 expression level according to the above aspects of the present invention or the method described in any one of the embodiments. In some embodiments, one form in which the operation guide may exist is printed information printed on a suitable medium or substrate (e.g., a piece of paper or several pieces of paper with information printed thereon), a kit package, etc. Another form in which the operation guide exists is a computer-readable medium on which information has been recorded, for example, a floppy disk, a compact disk (CD), a DVD, a portable flash drive, etc. Another form in which the operation guide may exist is a website, which can be used to access information on a remote website through the Internet.

[0020] In other aspects, the present invention provides a FKBP9 detection agent for liver cancer diagnosis, auxiliary diagnosis, screening and / or auxiliary screening and / or prognosis evaluation. In some embodiments, the liver cancer diagnosis, auxiliary diagnosis, screening and / or auxiliary screening and / or prognosis evaluation is implemented according to any one of the above aspects or embodiments of the present invention.

[0021] In other aspects, the present invention provides a composition comprising a FKBP9 detection agent, which is used for liver cancer diagnosis, auxiliary diagnosis, screening and / or auxiliary screening and / or prognosis evaluation. In some embodiments, the liver cancer diagnosis, auxiliary diagnosis, screening and / or auxiliary screening and / or prognosis evaluation is implemented according to any one of the above aspects or embodiments of the present invention.

[0022] In some embodiments, the determination in the above aspects of the present invention refers to a determination including quantitative and / or qualitative analysis, and includes detection of presence or absence and measurement of expression level. In some embodiments, the determination is qualitative or semi-quantitative, i.e., determining whether FKBP9 is present or measuring whether the concentration of FKBP9 is higher or lower than a reference value.

[0023] In some embodiments, the determination of FKBP9 expression level described in the above aspects of the present invention is to determine the level of FKBP9 gene expression products, wherein the KFBP9 gene expression products are polynucleotides, including DNA molecules and cDNA molecules, RNA molecules such as mRNA, and unspliced ​​or partially spliced ​​transcription products and spliced ​​products. In some embodiments, the gene expression level is determined by one or more methods of RNA-seq, RT-qPCR, qPCR, multiplex qPCR or RT-qPCR, microarray analysis, SAGE, MassARRAY technology, ISH. In some embodiments, the product for determining mRNA levels by PCR comprises mRNA-specific primers for FKBP9, and the product also includes a test tube or other suitable container, a reaction buffer, a deoxynucleotide (dNTP), an enzyme such as Taq polymerase and reverse transcriptase, a DNA enzyme, an RNA enzyme inhibitor, DEPC-water, sterile water, etc.

[0024] In some embodiments, the determination of FKBP9 expression level described in the above aspects of the present invention is to determine the level of FKBP9 protein expression products, wherein the FKBP9 protein expression products include polypeptides obtained by (complete or incomplete) transcription and translation of human FKBP9 genes. In some embodiments, the expression level of FKBP9 protein in a blood sample is determined by immunoassay, wherein the immunoassay utilizes the antigen-antibody reaction between FKBP9 antigen and anti-FKBP9 antibody. In some embodiments, the immunoassay is determined by one or more combinations of immunohistochemistry (IHC), Western blotting, enzyme-linked immunosorbent assay (ELISA), immunoprecipitation, immunofluorescence, radioimmunoassay, turbidimetric immunoassay, chemiluminescence or mass spectrometry.

[0025] In some embodiments, the FKBP9 detection agent described in the above aspects of the present invention is a molecule (e.g., an oligonucleotide) that can specifically detect the expression level of the FKBP9 gene or a molecule (e.g., an antibody or an antigen-binding fragment thereof) that can specifically detect the expression level of the FKBP9 protein. In some embodiments, the FKBP9 detection agent includes an anti-FKBP9 antibody, the antibody being selected from polyclonal antibodies and fragments thereof, monoclonal antibodies and fragments thereof, or a combination thereof, such as scFv, Fab, and F(ab)2 fragments, which can bind to FKBP9 antigens or haptens. In some embodiments, the FKBP9 detection agent is covalently or non-covalently coupled to a detectable label. Wherein labeling can be accomplished by direct or indirect methods. Direct labeling involves coupling the label directly (covalently or non-covalently) to the FKBP9 detection agent, and indirect labeling involves labeling the FKBP9 detection agent through an intermediate linking system (covalently or non-covalently), such as a secondary antibody with a detectable label to detect a primary antibody and a streptavidin protein-biotin system with a detectable label. In some embodiments, the detectable label is any substance that can be detected by fluorescence, spectroscopy, photochemistry, biochemistry, immunology, electricity, optics or chemical means, including but not limited to gold particles, latex beads, chemiluminescent substances, enzymatic activity labels, radioactive labels, magnetic labels and fluorescent labels. In some embodiments, the chemiluminescent substances are such as acridinium esters, luminol, ruthenium and its derivatives. In some embodiments, the enzymatic activity labels are such as horseradish peroxidase, alkaline phosphatase, β-galactosidase, luciferase and its derivatives, and suitable substrates for detection include diaminobenzidine (DAB), 3,3'-5,5'-tetramethylbenzidine, NBT-BCIP (4-nitro blue tetrazolium chloride and 5-bromo-4-chloro-3-indolyl-phosphate). In some embodiments, the fluorescent labels include fluorescent proteins (such as GFP and its derivatives), Cy3, Cy5, Texas Red, fluorescein and Alexa dyes (such as Alexa 568). In some embodiments, the magnetic bead labels are such as paramagnetic and superparamagnetic labels.

[0026] In some embodiments, the product described in the above aspects of the present invention is a biological detection kit, in particular an immunological detection kit, preferably an ELISA kit, wherein the ELISA kit is selected from a double antibody sandwich ELISA kit or a direct ELISA kit. In some embodiments, the ELISA kit comprises one or more of a solid phase carrier, a coating solution, an enzyme marker, a blocking solution, a sample diluent, an enzyme label diluent, a washing solution, a substrate, a reaction stop solution or a standard.

[0027] In some embodiments, the liver cancer described in the above aspects of the present invention is hepatocellular carcinoma. In some embodiments, the liver cancer is advanced liver cancer. In some embodiments, the liver cancer is advanced hepatocellular carcinoma. In some embodiments, the advanced liver cancer is unresectable liver cancer, including locally advanced or metastatic HCC. In some embodiments, the "unresectable" refers to a cancer that is impossible to be surgically resected or cannot be safely surgically resected, which is represented by the cancer being unable to be safely resected by partial hepatectomy, for example, because the tumor is too large to be safely resected, the part of the tumor located in the liver causes difficulty in removal (e.g., such as near large blood vessels), there are multiple tumors or cancer has spread to the entire liver and / or outside the liver, and / or the subject has a potential health problem (e.g., cirrhosis) that prevents resection.

[0028] In some embodiments, the blood sample described in the above aspects of the present invention is one or more of whole blood, serum or plasma, preferably one or more of serum and plasma.

[0029] In some embodiments, the individual, subject or patient is a mammal, such as a rodent, a feline, a canine, and a primate. Preferably, the individual, subject or patient is a human.

[0030] In some embodiments, the reference value is set by performing ROC analysis (Receiver Operating Characteristic analysis) on the FKBP9 expression level in the blood samples of liver cancer patients and / or the FKBP9 expression level in the blood samples of healthy control individuals. ROC analysis is an analysis method that can evaluate the detection and diagnostic capabilities of disease inspection methods, such as the analysis method described in: "The Japan Society for Clinical Laboratory Automation Journal "Evaluation Manual for Diagnostic Utility of Clinical Examination", Ver.1.3 (2004.9.1)), that is, the cutoff value is the reference value. In some embodiments, the reference value is obtained by performing ROC analysis on the FKBP9 protein expression level in the blood samples of liver cancer patients and / or the FKBP9 protein expression level in the blood samples of healthy control individuals.

[0031] In one embodiment, the cutoff value is determined according to sensitivity. For example, the sensitivity is at least or at most 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%. In one embodiment, the cutoff value is determined according to specificity. For example, the specificity is at least or at most 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%. In one embodiment, the cutoff value is determined according to sensitivity and specificity. In one embodiment, the cutoff value is determined according to AUC. For example, the AUC is at least 0.3, 0.35, 0.4, 0.45, 0.5, 0.55, 0.6, 0.65, 0.7, 0.75, 0.8, 0.85, 0.9, 0.95, or 1.

[0032] In some embodiments, the reference value may also be set to, for example, the sum of at least 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0 or more times the mean and standard deviation values ​​of the FKBP9 expression levels of healthy individuals and / or liver cancer patients, and may be further appropriately set to a value that satisfies sensitivity (detection rate) and specificity (the degree of low false positive rate) in a balanced manner. For example, the above scheme is applied by comparing the measured values ​​of FKBP9 protein in the serum of healthy individuals and the measured values ​​of FKBP9 protein in the serum of patients with advanced liver cancer, and further adjusting it to a value that satisfies sensitivity and specificity in a balanced manner. In some embodiments, the Youden index is used to balance sensitivity and specificity.

[0033] In some embodiments, the reference value may also be set to, for example, at least 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0 or more times the average value of the FKBP9 expression level of healthy individuals and / or liver cancer patients, and may be further appropriately set to a value that satisfies sensitivity (detection rate) and specificity (the degree of low false positive rate) in a balanced manner. For example, the above scheme is applied by comparing the measured value of FKBP9 protein in the serum of healthy individuals and the measured value of FKBP9 protein in the serum of patients with advanced liver cancer, and further adjusting it to a value that satisfies sensitivity and specificity in a balanced manner. In some embodiments, the Youden index is used to balance sensitivity and specificity.

[0034] In some embodiments, the median of FKBP9 gene expression or FKBP9 protein expression in blood samples of liver cancer patients can be used as a baseline value of its expression level, and the samples can be grouped into "high" and "low" groups based on this baseline value.

[0035] In some embodiments, the benchmark value can also be set as a cutoff value of the prognostic ROC. In some embodiments, the FKBP9 concentration when the Youden index is maximum is used as the cutoff value.

[0036] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0037] Unless otherwise indicated, the term "FKBP9" is short for FKBP prolyl isomerase 9 and refers to the human FKBP9 gene having the nucleotide sequence defined in NCBI Gene ID 11328 or the human FKBP9 protein having the amino acid sequence defined in NCBI GenPept O95302.2.

[0038] The term "biomarker" refers to a biological molecule whose presence, form or amount shows a statistically significant difference between two states. Biomarkers can be naturally occurring or non-naturally occurring, and can include but are not limited to nucleic acids, proteins, and variants and fragments thereof.

[0039] Beneficial effects: The present invention provides a biomarker for liver cancer diagnosis or prognosis assessment, and the use of FKBP9 detection agent in the preparation of liver cancer diagnosis, auxiliary diagnosis, screening, auxiliary screening, prognosis assessment, risk assessment and / or risk stratification products. The biomarker FKBP9 has a significant difference in blood content between healthy controls and patients with advanced liver cancer, and has high sensitivity and specificity, and can be used as a biomarker for diagnosing liver cancer. BRIEF DESCRIPTION OF THE DRAWINGS

[0040] Figure 1 To detect the expression level of serum FKBP9 using ELISA in advanced hepatocellular carcinoma group (aHCC) and healthy control group (HC); Figure 2 ROC curve diagram for distinguishing liver cancer patients from normal subjects based on FKBP9 expression level; Figure 3 Kaplan-Meier survival curves of HCC patients differentiated by FKBP9 expression levels. DETAILED DESCRIPTION

[0041] The technical solution of the present disclosure is further described below in conjunction with specific embodiments. However, the following embodiments are merely examples of the present disclosure and do not represent or limit the scope of protection of the present disclosure. In the following embodiments, unless otherwise specified, the reagents and consumables used are purchased from ordinary suppliers in the field, and the experimental methods and technical means used are conventional methods and means in the field.

[0042] Example 1: ELISA to detect the expression of FKBP9 in the serum of patients with advanced liver cancer and healthy controls

[0043] Collect 3-5mL whole blood samples from clinically confirmed hepatocellular carcinoma patients and healthy volunteers as controls. Centrifuge at 2-8℃, 1000×g for 15 minutes within 30 minutes after sample collection and take the supernatant. It can be tested immediately or after freezing and thawing at -20°C or -80℃.

[0044] The kit used to detect the FKBP9 content is the Human FKBP9 (Peptidyl-prolylcis-trans isomerase FKBP9) ELISA Kit (Cat. No.: EH5191) from FineTest®.

[0045] The results of the serum FKBP9 concentration test in liver cancer patients and healthy volunteers are shown in Table 1.

[0046] Table 1: FKBP9 detection results in serum of liver cancer patients and healthy volunteers

[0047] The results are as follows Figure 1 As shown in the results, the expression of FKBP9 was detected in the serum of both patients with advanced liver cancer and healthy controls. The expression level of FKBP9 in the serum of patients with advanced liver cancer (2.27±1.45 ng / mL) was significantly (P=0.0072) higher than that in healthy controls (1.31±0.62 ng / mL).

[0048] Draw the ROC curve, see Figure 2 The area under the ROC curve (AUC) of FKBP9 was 0.716. If the FKBP9 concentration (1.8773) at which the Youden index was the largest was selected as the critical value, the sensitivity was 57.1%, the specificity was 85.7%, and the Youden index was 0.428.

[0049] Table 2: Youden index calculation Positive if greater than or equal to this value Sensitivity 1 - Specificity Youden Index -0.6222 1 1 0 0.399 0.964 1 -0.036 0.4523 0.964 0.952 0.012 0.5313 0.929 0.952 -0.023 0.5901 0.929 0.905 0.024 0.6329 0.929 0.857 0.072 0.7025 0.929 0.81 0.119 0.7963 0.893 0.81 0.083 0.852 0.857 0.81 0.047 0.8573 0.857 0.762 0.095 0.8863 0.821 0.762 0.059 0.9225 0.786 0.762 0.024 0.9456 0.786 0.714 0.072 0.9749 0.786 0.667 0.119 1.0268 0.786 0.619 0.167 1.069 0.786 0.571 0.215 1.1309 0.75 0.571 0.179 1.2253 0.75 0.524 0.226 1.2705 0.75 0.476 0.274 1.3287 0.75 0.429 0.321 1.3859 0.75 0.381 0.369 1.3963 0.714 0.381 0.333 1.4249 0.714 0.333 0.381 1.4852 0.679 0.333 0.346 1.5373 0.643 0.333 0.31 1.5909 0.643 0.286 0.357 1.6887 0.607 0.286 0.321 1.7503 0.607 0.238 0.369 1.7867 0.607 0.19 0.417 1.823 0.571 0.19 0.381 1.8773 0.571 0.143 0.428 1.9504 0.536 0.143 0.393 1.9868 0.5 0.143 0.357 2.0098 0.464 0.143 0.321 2.0531 0.464 0.095 0.369 2.1135 0.429 0.095 0.334 2.1466 0.429 0.048 0.381 2.2292 0.393 0.048 0.345 2.3498 0.357 0.048 0.309 2.3998 0.321 0.048 0.273 2.6144 0.286 0.048 0.238 2.9101 0.25 0.048 0.202 3.0543 0.25 0 0.25 3.2252 0.214 0 0.214 3.3451 0.179 0 0.179 3.6806 0.143 0 0.143 4.612 0.107 0 0.107 5.35 0.071 0 0.071 5.6367 0.036 0 0.036 6.7878 0 0 0

[0050] The 28 HCC patients were divided into groups according to the cutoff value of serum FKBP9 (FKBP9 concentration when Youden index was the highest) of 1.8773 ng / mL, and the survival analysis curves were drawn. Figure 3The Kaplan-Meier method was used to estimate the survival function, and the Log-rank test was used to compare the differences in survival distribution between the high-expression and low-expression groups. The median survival time of patients with low serum FKBP9 expression (<1.8773 ng / mL) was greater than 36 months (not reached within the 3-year follow-up period), which was significantly longer than the median survival time of 10 months in patients with high expression (>1.8773 ng / mL) (P=0.037). Therefore, high serum FKBP9 expression in patients with liver cancer indicates a poor prognosis.

[0051] The above embodiment uses enzyme-linked immunosorbent assay to detect the serum FKBP9 content in healthy controls and patients with advanced liver cancer. After immunoanalysis of a large number of clinical samples, it is determined that there is a significant difference in the serum FKBP9 content between healthy controls and patients with advanced liver cancer, and ROC curve analysis is used to further verify that serum FKBP9 can be used as a serum biomarker for diagnosing liver cancer. It is also verified that FKBP9 in the blood can be used as a biomarker for the prognosis assessment of liver cancer. The present invention can achieve a better pre-diagnosis effect by blood sampling, which is of great significance for alleviating the sampling pain of the population to be tested and realizing a more popular screening.

[0052] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. Application of FKBP9 detection agents in the preparation of liver cancer diagnosis, auxiliary diagnosis, screening, auxiliary screening, prognosis evaluation, risk assessment and / or risk stratification products.

2. The use according to claim 1, characterized in that: The liver cancer diagnosis, auxiliary diagnosis, screening, and auxiliary screening include the following steps: (A1) determining the expression level of FKBP9 in a blood sample from a subject; (A2) FKBP9 expression levels were compared with baseline values; wherein an increased FKBP9 expression level compared to a baseline value indicates that the subject has liver cancer or is at risk of developing liver cancer; The prognostic assessment includes the following steps: (B1) Determining the expression level of FKBP9 in blood samples from liver cancer patients; (B2) FKBP9 expression levels were compared with baseline values; The increased FKBP9 expression level compared with the baseline value indicates that the liver cancer patient has a poor prognosis.

3. The use according to claim 1, characterized in that: The FKBP9 detection agent is a reagent for detecting the expression of the FKBP9 gene in a blood sample or a reagent for detecting the expression of the FKBP9 protein in a blood sample.

4. The use according to claim 3, characterized in that: The reagent for detecting the expression of FKBP9 gene in the blood sample is FKBP9 gene specific oligonucleotide.

5. The use according to claim 3, characterized in that: The reagent for detecting the expression of FKBP9 protein in the blood sample is an FKBP9 protein-specific antibody or an antigen-binding fragment thereof.

6. The use according to claim 2, characterized in that: The blood sample is one or more of whole blood, serum or plasma.

7. The use according to claim 2, characterized in that: The liver cancer is hepatocellular carcinoma, advanced liver cancer or one or more of advanced hepatocellular carcinoma.

8. The use according to claim 2, characterized in that: The method for determining the FKBP9 expression level is one or more of RNA-seq, RT-qPCR, qPCR, multiplex qPCR or RT-qPCR, microarray analysis, SAGE, MassARRAY technology, immunohistochemistry, Western blotting, enzyme-linked immunosorbent assay, immunoprecipitation, immunofluorescence, radioimmunoassay, turbidimetric immunoassay, chemiluminescence or mass spectrometry.

9. The use according to claim 2, characterized in that: The benchmark value is determined according to one or more of the following: sensitivity, specificity and / or AUC of the ROC curve; the mean value and / or median value of the FKBP9 expression level measured in blood samples from healthy people, and the optional standard deviation; and / or, the mean and / or median value of the FKBP9 expression level measured in blood samples from a group of liver cancer patients, and optionally the standard deviation.

10. The use according to claim 9, characterized in that: The reference value is: 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9 or 2.0 times the average value of the FKBP9 expression level measured in blood samples from healthy people; the average value of the FKBP9 expression level measured in blood samples from healthy people + 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9 or 2.0 times the standard value. Deviation; 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9 or 2.0 times the mean value of the FKBP9 expression level measured in blood samples from a group of liver cancer patients; the mean value of the FKBP9 expression level measured in blood samples from a group of liver cancer patients + 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9 or 2.0 times the standard deviation.

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