Transcriptome-sequencing-based gene group for early diagnosis of liver cancer and prognostic evaluation and application thereof

A technology for early diagnosis and early liver cancer, applied in the biological field, can solve the problem of low specificity and achieve the effects of reducing misdiagnosis rate, improving diagnosis, and improving detection ability and efficiency

Active Publication Date: 2017-08-18
上海东方肝胆外科医院
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Problems solved by technology

[0005] The present invention aims at the problem of low sensitivity and specificity of the current second-generation sequencing kits to carry out effective gene screening and combination, and uses the second-generation gene sequencing technology to screen out gene groups that are strongly associated with the diagnosis of primary liver cancer, which can significantly improve the Diagnosis of early primary liver cancer, reducing the rate of misdiagnosis, and at the same time, it is used to evaluate the prognosis of patients

Method used

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  • Transcriptome-sequencing-based gene group for early diagnosis of liver cancer and prognostic evaluation and application thereof
  • Transcriptome-sequencing-based gene group for early diagnosis of liver cancer and prognostic evaluation and application thereof

Examples

Experimental program
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Embodiment 1

[0056] Example 1 Screening of Gene Groups for Early Diagnosis and Prognostic Assessment of Primary Liver Cancer Based on Next Generation Sequencing Technology

[0057] From January 2008 to December 2008 in a tertiary hospital, a total of 200 patients with suspected liver cancer (single diameter ≤ 5 cm or the number of tumors less than 3, and all diameters ≤ 3 cm) were collected, and the postoperative diagnosis indicated focal tumors. There were 3 cases of nodular hyperplasia, 8 cases of atypical hyperplasia, and 2 cases of inflammatory nodules. There were 187 patients with early liver cancer and 13 cases of non-cancer patients. The isolated fresh tissues removed during the operation of the above 200 patients were made into paraffin tissue samples as required. Check as follows:

[0058] (1) Extraction of mRNA from paraffin tissue specimens (by QIAGEN RNeasy FFPE Kit)

[0059] 1) First trim the wax block with a blade;

[0060] 2) Cut into 5 μm wax slices on a paraffin microto...

Embodiment 2

[0122] Example 2 Diagnosis of early liver cancer and prognosis assessment based on gene group and RS value algorithm

[0123] Example 2 A total of 329 patients with early liver cancer (single diameter ≤ 5cm or number of tumors less than 3, and diameter ≤ 3cm) underwent surgery in a tertiary hospital from January 2010 to January 2011 were collected, and the postoperative diagnosis indicated localized liver cancer. There were 3 cases of focal nodular hyperplasia, 7 cases of atypical hyperplasia, and 4 cases of inflammatory nodules. According to the requirements, the isolated fresh tissue taken out of the patient's operation was made into a paraffin tissue sample, and the total RNA of the sample was extracted, reverse-transcribed into cDNA, and six GPC3, HSP70, GS, PIVKA-Ⅱ, GAPDH, and ACTB were detected in the sample by qPCR The expression level of the gene was calculated according to the RS value calculation formula in Example 1 to obtain the RS value of each patient. Comparing...

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Abstract

The invention discloses a transcriptome-sequencing-based gene group for early diagnosis of liver cancer and prognostic evaluation and application thereof; the gene group comprises four genes, GPC3, HSP70, GS and PIVKA-II. Recurrence risk value is calculated according to expression amounts of the four genes, whether a patient under test carries early liver cancer or not and prognostic recurrence rate of a patient with liver cancer are judged according to the recurrence risk value. An algorithm to calculate the recurrence risk value via the expression amounts is RS=0.080*GP3+0.098*HSP70+0.047*GS+0.13*PIVKA-II. Preclinical verification results are precise, diagnostic sensitivity for early primary liver cancer reaches 100%, and diagnostic sensitivity for non-liver cancers reaches 100%, prediction sensitivity to a low-recurring risk patient reaches 100%, prediction sensitivity to a medium-recurring risk patient reaches 96.774%, prediction sensitivity to a high-recurring risk patient reaches 99.375%, and detection capacity and efficiency are improved with detection cost greatly reduced; clear tips can be provided upon a patient's prognosis.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a gene group for early diagnosis and prognosis evaluation of liver cancer based on transcriptome sequencing technology and its application. Background technique [0002] Primary hepatocellular carcinoma (Hepatocellular Carcinoma, HCC, referred to as liver cancer) is the most malignant tumor, its incidence rate ranks sixth in the world, and its case fatality rate ranks third. The comprehensive treatment model based on surgery has achieved remarkable results in the treatment of liver cancer. However, since patients with liver cancer are often found in the middle and advanced stages and accompanied by different degrees of cirrhosis, the surgical resection rate is low, the recurrence rate is high, and the prognosis is poor. At present, protein serum marker screening and imaging diagnosis are the main means to detect residual tumor, recurrence and metastatic lesions in patient...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6869C12Q1/6886C12Q2600/118C12Q2600/158C12Q2535/122
Inventor 杨田李超梁磊邢昊韩骏张汉
Owner 上海东方肝胆外科医院
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