Group of genes for prognosis evaluation of adrenocortical carcinoma

Abstract

The invention discloses a group of genes for prognosis evaluation of adrenocortical carcinoma, and belongs to the field of biological and medical examination. The related genes comprise CCNB2, CDCA5, MCM10 and TACC3 (Triglyceride 3). The molecular typing and prognosis evaluation are performed on the adrenal cortical carcinoma by detecting the expression quantity of products (RNA and protein) of the genes (single gene or any combination); and a reliable method is provided for rapidly predicting the molecular typing and prognosis of the adrenal cortex cancer.

Description

[0001] This application is a divisional application of the invention titled "Gene Expression Profile for Prognosis of Adrenocortical Carcinoma", with an application date of December 22, 2020 and an application number of 202011531814.X. Technical Field

[0002] 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 levels of single or any combination of related gene products (proteins, RNAs) 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

[0003] Adrenocortical carcinoma (ACC) is a tumor with a low incidence but high invasiveness, and the prognosis of some patients is poor. Rapidly differentiating the malignancy and prognostic molecular typing of adrenocortical carcinoma is of great value for the selection of treatment regimens. In the present invention, through large-scale screening of the relationship between the gene expression profile of adrenocortical carcinoma and prognosis, a group of genes related to the prognosis of adrenocortical carcinoma was found: 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. The combined application of the above genes is beneficial to the prognosis determination and molecular typing of adrenocortical carcinoma.

[0004] Based on the research foundation and current situation of the existing technology, the inventors of the present application intend to provide an index for the prognosis and molecular typing of adrenocortical carcinoma. The present invention relates to predicting the malignancy and prognosis of adrenocortical carcinoma by the high or low expression levels of the above-mentioned genomes (single or any combination of two or more) in adrenocortical carcinoma tissues. 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, fluorescence quantitative PCR, RNA array, RNA Sequencing, etc.) in adrenocortical carcinoma tissues, 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

[0005] 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 profiles of related genes.

[0006] Based on the analysis of large-scale gene expression profiles, the present invention screens out a group of genes highly related to the prognosis of adrenocortical carcinoma, including: 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, UBE2T and other 29 genes. The molecular typing and prognosis evaluation of the above-mentioned adrenocortical carcinoma are related to the expression levels of the above genes (single or any combination) in adrenocortical carcinoma, and can be used for predicting the molecular typing, drug resistance, drug screening, patient prognosis analysis, etc. of adrenocortical carcinoma;

[0007] S1 Use the above gene expression levels alone to judge the molecular typing and patient prognosis of adrenocortical carcinoma, etc.;

[0008] S2 Combine the changes in the expression levels of the above genes (any combination of 2 or more) to judge the molecular typing and patient prognosis of adrenocortical carcinoma, etc.;

[0009] S3 After combining the above single gene or any combination with other indicators, judge the molecular typing and patient prognosis of adrenocortical carcinoma, etc.;

[0010] The present invention relates to detecting the above gene expression products including: proteins and RNA.

[0011] The above-mentioned gene products involved in the present invention include those in tumor cells, as well as mRNA, proteins, and exosomes that enter body fluids for various reasons, including both complete and fragmented ones.

[0012] The detection objects of the present invention include: tumor tissues, body fluids (including blood, urine, puncture fluids, etc.), puncture tissues, etc., which are used as molecular typing of adrenocortical carcinoma and evaluation indicators for patients, etc.

[0013] The body fluids involved in the present invention include blood, urine, tumor puncture fluids, lymphatic fluids, etc.

[0014] The blood involved in the present invention includes whole blood, serum, plasma, isolated nucleated cells, circulating tumor cells in blood, exosomes, etc.

[0015] The above-mentioned gene protein detection involved in the present invention includes directly extracting the protein of adrenocortical carcinoma tissue and detecting and immunohistochemically analyzing it by chemical or immunological methods.

[0016] The above-mentioned gene RNA detection involved in the present invention includes detecting by directly extracting the RNA of adrenocortical carcinoma tissue and then combining RT-PCR technology, fluorescence quantitative PCR technology, RNA Sequencing, RNA array, etc.

[0017] The detection of gene products involved in the present invention includes protein level and RNA level, and the detection methods involve: immunohistochemistry, immunofluorescence, Western blot, ELISA, flow cytometry detection, RT-PCR, colloidal gold, immunological detection, chemical detection, fluorescence quantitative PCR, RNA Sequencing, RNA array and other detection technologies (but not limited to these technologies), etc.

[0018] The RT-PCR involved in the present invention detects the RNA products of the above-mentioned genes. After reverse transcription, the PCR involved includes: ordinary PCR, fluorescence quantitative PCR, nested PCR, multiplex PCR, digital PCR, etc.

[0019] The beneficial effects of the present invention are:

[0020] Microscale, rapidity, and high throughput are the general trends of technological development; although pathological morphology is the gold standard for diagnosing tumors, its effect in the prognostic evaluation of tumors is limited. Therefore, molecular biological indicators are expected to become important means for tumor prognostic evaluation and molecular typing. In the present invention, we provide a molecular method for molecular typing, drug screening, prognostic evaluation, etc. of adrenocortical carcinoma based on changes in the tumor genome expression profile. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1.The heatmap of the mRNA Sequencing results shows the overall expression of the mRNAs of 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 adrenocortical carcinoma (ACC) tissues.

[0022] Figure 2 .The heatmap of the mRNA Sequencing results shows the overall expression of the mRNAs of 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 adrenocortical carcinoma (ACC) tissues.

[0023] Figure 3 .The survival curve results show the relationship between the mRNA expression levels of genes ASPM, C3, CCNB2, CDCA3, CDCA5, CEP95 (CCDC45), CENPM, CENPW, DNA2, DNAJC9, ECT2, EME1, FANCI, G0S2, HAUS8, HELLS, etc. and the overall survival time of patients with adrenocortical carcinoma (ACC). Among them, those with high expression levels of genes ASPM, CCNB2, CDCA3, CDCA5, CEP95, CENPM, CENPW, DNA2, DNAJC9, ECT2, EME1, FANCI, HAUS8, HELLS, etc. generally have a higher degree of tumor malignancy and a poorer prognosis; while those with higher expression levels of genes C3, G0S2, etc. have a lower degree of tumor malignancy and a better prognosis.

[0024] Figure 4 .The survival curve results show the relationship between the mRNA expression levels of genes KIF11, LMO7, MCM10, MXD3, NUP188, NVL, PKMYT1, PSRC1, TACC3, TOMM5, TRIP13, UBE2T, H1F0, etc. and the overall survival time of patients with adrenocortical carcinoma (ACC). Those with high expression levels generally have a higher degree of tumor malignancy and a poorer prognosis.

[0025] Figure 5 The ROC index and AUC data of the CCNB2 gene are shown.

[0026] Figure 6 Displays the ROC index and AUC data of the CDCA5 gene.

[0027] Figure 7 Displays the ROC index and AUC data of the MCM10 gene.

[0028] Figure 8 Displays the ROC index and AUC data of the TACC3 gene. Detailed implementation mode

[0029] Example 1

[0030] In the present invention, we take adrenocortical carcinoma (ACC) as the implementation object, and take the analysis of the relationship between the mRNA expression level in adrenocortical carcinoma tissues and the prognosis of patients by RNA Sequencing technology as an example. Based on the study of the relationship between the mRNA expression level of genes in adrenocortical carcinoma tissues and prognosis in the The Cancer Genome Atlas (TCGA) database, we screened a group of genes closely related to the prognosis of patients, 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).

[0031] Through the study of the relationship between the mRNA Sequencing results in the TCGA database and the overall survival curve of patients, we found that: the expression levels of genes such as 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 indicate that the malignancy of adrenocortical carcinoma is generally higher and the prognosis of patients is poorer; while the higher expression levels of genes such as C3 and G0S2 indicate that the malignancy of adrenocortical carcinoma is lower and the prognosis of patients is better ( Figure 3 , 4).

[0032] In the present invention, the results of mRNA Sequencing in the TCGA database show that there are also differences in the above gene expression profiles. Molecular typing of adrenocortical carcinoma can be carried out based on the differences in the above gene expression profiles; for example: ASPM high / CCNB2 high / CDCA3 high / CDCA5 high type has a higher malignancy and a poorer prognosis; ANXA8 low / LGALS7B low / CALML3 low / PKP1 low type has a lower malignancy and a better prognosis, etc., and adrenocortical carcinoma is divided into different molecular subtypes ( Figure 1 , 2).

[0033] Summary

[0034] Our results show that 29 genes such as 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 are closely related to adrenocortical carcinoma, and the levels of mRNA expression of the above genomes can be used for molecular typing and prognosis evaluation of adrenocortical carcinoma. Hint: The present invention provides a reliable marker for molecular typing and prognosis evaluation of adrenocortical carcinoma.

Claims

1. A group of genes for prognostic evaluation of adrenocortical carcinoma (ACC), and the genes involved include: CCNB2, CDCA5, MCM10 and TACC3 are used to perform molecular typing, malignancy grading, survival analysis and prognosis assessment of adrenocortical carcinoma by detecting the expression levels of the above gene products in biological samples. S1: Use the above gene expression levels alone to determine the molecular typing, malignancy grading, survival analysis and patient prognosis of adrenocortical carcinoma; S2: Combine the changes in the above gene expression levels to determine the molecular typing, malignancy grading, survival analysis and patient prognosis of adrenocortical carcinoma; S3: After combining the above single genes or any combination and then combining with other indicators, determine the molecular typing, malignancy grading, survival analysis and patient prognosis of adrenocortical carcinoma.

2. The use according to claim 1, characterized in that, The detection objects include: tumor tissues, body fluids, and puncture tissues, which are used as indicators for molecular typing and survival prognosis assessment of adrenocortical carcinoma.

3. The use according to claim 1, characterized in that, The detected gene products include: proteins and RNAs, including those in tumor cells, as well as free RNAs, proteins and RNAs and proteins in exosomes that enter body fluids for various reasons, including complete and fragmented ones.

4. The use according to claim 1 or 2, characterized in that, The blood involved includes whole blood, serum, plasma, isolated nucleated cells, circulating tumor cells in blood, and exosomes in blood.

5. The use according to claim 1, characterized in that, Protein concentration detection includes directly extracting proteins from adrenocortical carcinoma tissues and detecting them by chemical or immunological methods and immunohistochemical analysis. The RNA concentration detection includes directly extracting RNAs from adrenocortical carcinoma tissues for detection.

6. The use according to claim 1, 3 or 5, characterized in that, The detected genes include: gene expression detection includes protein level and RNA level. The detection methods involve: immunohistochemistry, immunofluorescence, Western blot, ELISA, flow cytometry detection, RT-PCR, immunological detection, colloidal gold, chemical detection, fluorescence quantitative PCR, RNA Sequencing or RNAarray.

7. The use according to claim 1, characterized in that, Directly detect the absolute amount of the above genes; or the ratio to normal tissues, and use it as an evaluation of the molecular typing, malignancy grading, and survival malignancy indicators of adrenocortical carcinoma.

8. The use according to claim 1 or 6, characterized in that, The RT-PCR mentioned above detects the RNA products of the above genes. After reverse transcription, the PCR involved includes: ordinary PCR, fluorescence quantitative PCR, nested PCR, multiplex PCR, digital PCR or chip microarray.

9. The use according to claim 1, characterized in that, Form a tumor marker matrix or chip pattern through the combined application of the above genes to improve the efficiency of molecular typing and prognosis judgment of adrenocortical carcinoma.

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