New mutation of hMLH1 gene of colorectal cancer patients in north China and use thereof

A colorectal cancer and genetic technology, applied in the direction of genetic engineering, plant gene improvement, application, etc., can solve the problem of unreported mutations

Inactive Publication Date: 2012-08-22
HARBIN MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the mutation of hMLH1 gene in patients with sporadic colorectal cancer in northern China has not been reported, especially the newly discovered mutation (c.704GAT>GTT(p.Asp235Val)) in a patient with sporadic colorectal cancer has not been reported at home and abroad

Method used

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  • New mutation of hMLH1 gene of colorectal cancer patients in north China and use thereof
  • New mutation of hMLH1 gene of colorectal cancer patients in north China and use thereof
  • New mutation of hMLH1 gene of colorectal cancer patients in north China and use thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0035] (1) DNA extraction from tissue samples by saturated phenol chloroform method:

[0036] ① Grind the resected rectal cancer tissue of a sporadic colorectal cancer patient with liquid nitrogen, add 4ml TES buffer solution, 250μl 10% SDS solution, 50μl 20mg / ml proteinase K solution to the ground tissue powder, mix well, Disperse the aggregated leukocytes; keep the temperature in a water bath at 56°C for 2 hours.

[0037] ② Add an equal volume of saturated phenol, repeatedly and slowly invert the centrifuge tube, and mix the two phases to form an emulsion.

[0038] ③ Centrifuge at 5000 rpm, 4°C, for 15 minutes.

[0039] ④ Use a large-bore straw to absorb the viscous aqueous phase (upper layer) and transfer it to another clean centrifuge tube.

[0040] ⑤ Repeat steps ②, ③, ④.

[0041] ⑥ Add an equal volume of chloroform-isoamyl alcohol (volume ratio: 24:1) solution, slowly invert the centrifuge tube repeatedly, and mix the two phases to form an emulsion. Repeat steps ③, ④...

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Abstract

The invention discloses new mutation of hMLH1 gene of colorectal cancer patients in north China and use of the hMLH1 gene, and relates to a new mutation type of the hMLH1 gene of the sporadic colorectal cancer patients in north China. According to the invention, the new mutation is discovered in the 1 / 324 sporadic colorectal cancer patients, i.e. c.704GAT is greater than GTT (glucose tolerance test) (p.Asp235Val). The invention comprises the following operation steps of: 1. extracting DNA (deoxyribonucleic acid); 2, designing a primer; 3, carrying out PCR-SSCP (polymerase chain reaction-single strand conformation polymorphism); 4, sequencing; and 5, checking with the GeneBank original sequence. The mutation spectrum of the hMLH1 gene in the colorectal cancer patients is more enriched on the basis of the reported hMLH1 gene mutation. The theoretical basis is provided for the molecular diagnosis and the treatment of the colorectal cancer patients.

Description

technical field [0001] The invention relates to a new mutation type of hMLH1 gene in patients with sporadic colorectal cancer in northern China. It belongs to the field of molecular detection. Background technique [0002] The hMLH1 gene is one of the pathogenic genes in patients with colorectal cancer. Since Papadopoulos N et al. discovered the hMLH1 gene mutation in Lynch syndrome colorectal cancer patients in 1993, most of the literature at home and abroad have reported the mutation site of hMLH1 gene in Lynch syndrome colorectal cancer, and only a few literatures have reported hMLH1 gene mutation sites in Lynch syndrome colorectal cancer patients. Mutation sites in sporadic colorectal cancer. However, the mutation of hMLH1 gene in patients with sporadic colorectal cancer in northern China has not been reported, especially the newly discovered mutation (c.704GAT>GTT(p.Asp235Val)) in a patient with sporadic colorectal cancer has not been reported at home and abroad. ...

Claims

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Application Information

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IPC IPC(8): C12N15/12C12N15/11C12Q1/68
Inventor 胡付兰王帆李丹丹王彝白纳姚晓萍
Owner HARBIN MEDICAL UNIVERSITY
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