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Methylation gene for liver cancer screening

A methylation and gene technology, applied in the fields of biochemistry and molecular biology, can solve problems such as poor detection of early liver cancer, and achieve the effects of strong practicability, reliable results and high sensitivity

Active Publication Date: 2017-06-09
BEIJING YOUAN HOSPITAL CAPITAL MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The method is also ineffective in detecting early-stage liver cancer

Method used

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  • Methylation gene for liver cancer screening
  • Methylation gene for liver cancer screening
  • Methylation gene for liver cancer screening

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1 Construction of a clinical data sample resource bank for hepatitis B-related liver cancer

[0025] Inclusion criteria: age 18-70 years old; gender is not limited; the diagnostic criteria of hepatitis B-related liver cancer are based on the 2012 EASL guidelines EASL-EORTC Clinical Practice Guidelines: Management of hepatitis B virus chronic infection, CT or MRI four-phase enhanced scan showed Typical arterial perfusion rapid outflow, pathological examination is the gold standard); liver cancer staging standard, primary liver cancer staging is based on the 2012 EASL guidelines EASL-EORTC Clinical Practice Guidelines: Management of hepatocellular carcinoma (Barcelona criteria), where stage 0 is defined as Early stage and stage A are defined as early liver cancer, stage B is defined as advanced liver cancer, and stages C and D are defined as advanced liver cancer.

[0026] Exclusion criteria: non-hepatitis B-related liver disease or liver cancer; pregnant women or...

Embodiment 2

[0036] Example 2 Study on the difference of DNA methylation profiles of T cells corresponding to different clinical stages of liver cancer

[0037] 1) Determine the DNA methylation profile related to the immune function of candidate T cells:

[0038] a) Establish T cell DNA methylation profile: Based on the current research progress of DNA methylation, from the disease-related methylation profile, determine the pathways that affect T cell toxicity, antigen presentation, and T cell immune activation More than 30,000 sites in DNA methylation profiles;

[0039] b) Establish a high-throughput liquid phase chip detection platform for DNA methylation in T cells: use real-time PCR technology to amplify candidate genes, and use pyrosequencing to determine DNA methylation changes in candidate genes;

[0040] c) Establish a T cell DNA methylation analysis technology platform: use biological-computing analysis technology to perform cluster analysis on data.

[0041] 2) Study on the dif...

Embodiment 3

[0048] Example 3 Methylation site detection - nest PCR (STAP1)

[0049] out Nest PCR system preparation:

[0050]

[0051] PCR program: 95°C 5min; 95°C 30s, 56°C 30s, 72°C 30s, 5cycle; 95°C 30s, 54°C 30s, 72°C 30s, 5cycle; 95°C 30s, 52°C 30s, 72°C 30s, 10cycle; 95°C ℃30s, 50℃30s, 72℃30s, 15cycle; 72℃5min; 10℃forever.

[0052] Inner Nest PCR system preparation

[0053]

[0054] PCR program: 95°C 5min; 95°C 30s, 56°C 30s, 72°C 30s, 5cycle; 95°C 30s, 54°C 30s, 72°C 30s, 5cycle; 95°C 30s, 52°C 30s, 72°C 30s, 10cycle; 95°C ℃30s, 50℃30s, 72℃30s, 15cycle; 72℃5min; 10℃forever.

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Abstract

The invention discloses a methylation gene for liver cancer screening. The gene is STAP1, and the nucleotide sequence of the gene is as shown in SEQ ID No: 1 and at least contains a methylation locus. According to the methylation gene for liver cancer screening, methylation sequencing can be performed by the aid of methods such as a methylation specific polymerase chain reaction method, a bisulfite sequencing method, a micro-array method, a mass-spectrographic analysis method, a denaturing high-performance liquid chromatography method and a pyrosequencing method, and the methylation gene is high in sensitivity, reliable in result and high in practicability and can sensitively detect early-phase liver cancer and earlier-phase liver cancer.

Description

technical field [0001] The invention belongs to the technical field of biochemistry and molecular biology, and specifically relates to a methylation gene used for liver cancer screening. Background technique [0002] Liver cancer is one of the common malignant tumors in my country. According to statistics, the annual death rate of liver cancer in my country is 20.37 / 100,000, ranking second in the death sequence of malignant tumors, second only to lung cancer in cities and gastric cancer in rural areas. Liver cancer can be divided into liver cirrhosis type, liver abscess type, liver tumor type, and cancer metastasis type according to clinical manifestations. Among the above types, liver tumor type is the most common. About half of the patients complained of upper abdominal mass, followed by liver abscess type. About 1 / 3 of the cases had upper abdominal pain and hepatomegaly. Both epidemiological and experimental studies have shown that viral hepatitis has a specific relatio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6886C12Q2600/154
Inventor 李宁张永宏李康刘桂海
Owner BEIJING YOUAN HOSPITAL CAPITAL MEDICAL UNIV
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