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Molecular marker GRIK3 in rectal adenocarcinoma

A technology for rectal adenocarcinoma and expression products, which can be applied in the biological field and can solve the problem of low five-year survival rate

Active Publication Date: 2019-11-05
成都望路医药技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Patients with rectal adenocarcinoma have significantly poor five-year survival rates even with surgical resection

Method used

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  • Molecular marker GRIK3 in rectal adenocarcinoma
  • Molecular marker GRIK3 in rectal adenocarcinoma
  • Molecular marker GRIK3 in rectal adenocarcinoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Example 1 Screening for gene markers associated with rectal adenocarcinoma

[0056] 1. Collection of samples

[0057] Eight cases of rectal adenocarcinoma tissue and adjacent tissue samples of rectal adenocarcinoma were collected, and all the above specimens were obtained with the consent of the organizational ethics committee. All surgical resection specimens were cut into small pieces with sterilized ophthalmic scissors, put into 1.5ml sterile EP tubes soaked in 0.1% DEPC water overnight and sterilized, and the remaining tissues were stored in cryopreservation tubes and quickly put into Store in a liquid nitrogen tank at -80°C.

[0058] 2. Preparation of RNA samples

[0059] 1) Sample pretreatment: Take the sample from the -80°C liquid nitrogen tank and put it into a homogenizer soaked in 0.1% DEPC water overnight and sterilized, and grind it into powder; during the grinding process, the sample must always be in the In a liquid nitrogen environment, the grinding cu...

Embodiment 2

[0079] Example 2 QPCR sequencing to verify the differential expression of GRIK3 gene

[0080] 1. According to the detection results of high-throughput sequencing, the GRIK3 gene was selected for large-sample QPCR verification. According to the sample collection method in Example 1, 70 cases of rectal adenocarcinoma paracancerous tissues and 70 cases of rectal adenocarcinoma tissues were selected.

[0081] 2. The RNA extraction steps are the same as in Example 1.

[0082] 3. Reverse transcription:

[0083] 1) Reaction system:

[0084] Reagent volume MgCl 2

2μl 10×RT Buffer 1μl RNase-free water 3.75μl dNTP mix 1μl RNase inhibitor 0.25μl AMV reverse transcriptase 0.5μl Random 9mers 0.5μl experimental sample RNA 1μl

[0085] 2) Reverse transcription reaction conditions

[0086] According to the reverse transcription reaction conditions in RNA PCR Kit (AMV) Ver.3.0.

[0087] 30°C for 10min, 50°C for 25...

Embodiment 3

[0108] Embodiment 3 inhibits GRIK3 gene expression

[0109] 1. Cell culture (human rectal adenocarcinoma cell line HRC-99):

[0110] 1) Cell recovery

[0111] a. Take out the cell cryopreservation tube from the liquid nitrogen tank, immediately put the cell cryopreservation tube into warm water at 37°C, and shake it gently to melt it as soon as possible.

[0112] b. Take out the cryopreservation tube from warm water at 37°C, suck out the cell suspension with a straw, add 5ml of RPMI1640 culture medium containing 10% fetal bovine serum to the centrifuge tube, mix and centrifuge at 800rpm for 5min, remove the supernatant, and use Culture medium was washed once.

[0113] c. Add 5ml of RPMI1640 culture solution containing 10% fetal bovine serum, inoculate in a 75ml culture bottle, place at 37°C, 5% CO 2 cultured in a constant temperature incubator.

[0114] 2) Cell replacement

[0115] a. When the culture solution starts to turn yellow, take out the culture bottle from the in...

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Abstract

The invention discloses a molecular marker GRIK3 of rectal gland cancer. Experiments prove that GRIK3 has high expression in rectal gland cancer tissue, and rectal gland cancer can be diagnosed by detecting the expression level of GRIK3. The invention also discloses an application of GRIK3 genes in preparation of a medicine for treating rectal gland cancer.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the molecular marker GRIK3 of rectal adenocarcinoma. Background technique [0002] Rectal adenocarcinoma is one of the common malignant tumors of the digestive tract. In the United States and Western countries, its incidence rate ranks third among malignant tumors, and it is also the third most common cause of death caused by tumors. In China, its morbidity and mortality are at the forefront of various malignant tumors, and it is increasing year by year, which may be related to the following reasons: (1) The adjustment of dietary structure and the improvement of people's quality of life The animal fat intake The increase of protein increases the amount of methylcholanthrene and other substances that can induce rectal cancer in the feces, and the intake of dietary fiber that can increase the volume of the stool, speed up the passage of the stool and reduce the absorption of harmful s...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K45/00A61P35/00C12Q1/6886G01N33/68
CPCA61K45/00C12Q1/6886C12Q2600/158G01N33/57446G01N33/6893
Inventor 肖枫宋宏涛杨承刚
Owner 成都望路医药技术有限公司
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