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Molecular marker related to gastric adenocarcinoma metastasis

A technology of gastric adenocarcinoma and drugs, which is applied in the field of tumor diagnosis by means of detection of PPM1A abnormality, and can solve problems such as unclear molecular mechanisms

Active Publication Date: 2017-11-17
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, for advanced gastric cancer, the underlying molecular mechanism of invasion and metastasis is still unclear.

Method used

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  • Molecular marker related to gastric adenocarcinoma metastasis
  • Molecular marker related to gastric adenocarcinoma metastasis
  • Molecular marker related to gastric adenocarcinoma metastasis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Example 1 Differential expression of PPM1A gene

[0064] 1. Sample collection

[0065] 50 cases of metastatic gastric adenocarcinoma tissues and their adjacent normal tissues were collected, and 45 cases of non-metastatic gastric adenocarcinoma tissues and their adjacent normal tissues were collected. The tissue samples were collected with the help of pathologists, and the sample collection standards were as follows: (1) primary gastric adenocarcinoma, no other diseases, and patients who had not undergone cancer radiotherapy and chemotherapy before surgery; (2) samples diagnosed as gastric adenocarcinoma by the pathology department, and the cancer (3) In order to avoid unnecessary cross-contamination, two sets of equipment were used for sampling. First, samples of normal tissue adjacent to cancer that were at least 5 cm away from the cancer tissue without obvious lesions were taken. For full-thickness gastric adenocarcinoma tissue samples, immediately after collection,...

Embodiment 2

[0111] Example 2 Differential expression of PPM1A protein

[0112] 1. Extract the total protein of the tissue sample in Example 1

[0113] Follow the instructions of the EpiQuik Tissue / Cell Total Protein Extraction Kit for protein extraction.

[0114] 2. Western blot

[0115] β-actin was used as internal reference. 50 μg of total protein was separated by SDS-PAGE, electrotransferred to PVDF membrane, and blocked with 1×TBST containing 5% skimmed milk powder at room temperature for 1 h; added rabbit anti-human PPM1A monoclonal antibody (1:800 dilution) and mouse anti-human β-actin polyclonal antibody (diluted 1:3 000), overnight at 4°C; wash the membrane 4 times with 1×TBST, add goat anti-rabbit and goat anti-mouse IgG (diluted 1:2 000), incubate at room temperature for 1 h; 1×TBST After washing the membrane 4 times, put it in Super Signal chemiluminescent reagent to react for 2 minutes, expose the X-ray film in the dark room, develop and fix it by conventional method.

[0...

Embodiment 3

[0120] Example 3 Overexpression of PPM1A gene

[0121] 1. Plasmid construction

[0122]Amplification primers are designed according to the coding sequence of PPM1A gene, and the design of primers is well known to those skilled in the art. Amplify the coding sequence of the full-length PPM1A gene from the cDNA library of adult fetal brain (clontech company, article number: 638831), insert the above cDNA sequence into the eukaryotic cell expression vector pcDNA3.1, and connect the obtained recombinant vector pcDNA3.1 -PPM1A was used in subsequent experiments.

[0123] 2. Culture of human gastric adenocarcinoma cells

[0124] Human gastric adenocarcinoma SGC7901 cells were treated with RPMI1640 medium containing 10% fetal bovine serum plus penicillin 100units / ml and streptomycin 100μg / ml, placed at 37°C, 5% CO 2 Cultured in an incubator, the culture medium was changed every 24 hours, and the cells were passaged once every 48 hours. Cells in the logarithmic growth phase were u...

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Abstract

The invention discloses a molecular marker-PPM1A gene related to gastric adenocarcinoma metastasis. Through detection of the contents of PPM1A gene and expression products of the PPM1A gene in tissue of a subject, whether or not the subject suffers from gastric adenocarcinoma is judged or the subject has a risk of suffering from the gastric adenocarcinoma is diagnosed. Moreover, through study of invasion and migration indexes of gastric adenocarcinoma cells cultured in vitro, the PPM1A gene being as a drug target for curing the gastric adenocarcinoma is proved.

Description

technical field [0001] The present invention relates to the field of tumor diagnosis and treatment, more specifically, the present invention relates to a tumor diagnosis method by means of detecting PPM1A abnormality; and a tumor therapeutic agent promoting PPM1A gene or protein. Background technique [0002] Gastric cancer is a malignant tumor derived from gastric mucosal epithelial cells, mainly adenocarcinoma. In 2008, nearly 1 million cases of gastric cancer were newly diagnosed in the world, and the number of deaths was 740,000. It is the fourth most common cancer in the world and the second most lethal. Although the overall global incidence of gastric cancer has declined, nearly two-thirds of gastric cancer cases are concentrated in economically underdeveloped countries and regions, with high incidence in China and other East Asian countries. According to the latest version of "China Cancer Map" released by "Life Times" in 2014. There are about 3,120,000 new cancer c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N33/574A61K45/00A61P35/04
CPCA61K45/00C12Q1/6886C12Q2600/118C12Q2600/136C12Q2600/158G01N33/57446G01N33/57484G01N2800/52G01N2800/54
Inventor 宋宏涛马翠
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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