Method for inducing cell malignant transformation culture model

Abstract

The invention relates to a method for inducing a cell malignant transformation culture model. The method comprises the following steps: (1) culturing immortalized human gastric mucosal epithelial GES-1 in a DMEM culture solution containing 10% fetal bovine serum to obtain cultured cells; (2) culturing Cag A-positive helicobacter pylori J99 in a brain heart infusion medium containing 10% of newborncalf serum and centrifuging to remove a supernatant to obtain cultured bacteria; (3) diluting the cultured bacteria with DMEM, adding the bacteria into the cultured cells for induction by inoculationevery other day, adding an MNNG solution daily, and obtaining an induced helicobacter pylori infection model 48 weeks later. The method is simple and convenient, not only various indexes of malignanttransformation of the cells can be observed, but also the cell model in which the helicobacter pylori and MNNG work together for long-term for malignant transformation of gastric mucosa can be established, and a good experiment foundation is laid for study of the subsequent gastric cancer formation mechanism and the development of cancer treatment drugs.

Description

technical field [0001] The invention relates to the technical field of basic medical experiments, in particular to a method for inducing a cell malignant transformation culture model. Background technique [0002] In 1994, the World Health Organization (WHO) defined Helicobacter pylori as a class of carcinogens. The global infection rate is close to 50%. When Helicobacter pylori continues to infect the gastric mucosa for a long time, it will cause gastric diseases, including chronic active gastritis, dyspepsia, gastric ulcer, and even gastric cancer and gastric mucosa-associated lymphoma (MALT lymphoma). Another carcinogen, nitroso compounds (N-nitroso compounds, NOCs) widely exists in the environment of human life, such as food, cigarettes or occupational factors. This long-term exposure to nitroso compounds can cause gastric cancer. my country is a country with a high incidence of gastric cancer. New gastric cancers in China account for 42% of new gastric cancers in the ...

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Problems solved by technology

The model of MNNG-induced human gastric mucosal cells is usually also used to detect the indicators of the cells after 24 hours (Yan, Z.P., et al., [Injury of human gastric epithelial GES-1 cells by MNNG and its effects on Wnt / beta-catenin signaling pathway]. Sheng Li Xue Bao, 2018. 70(3): p. 262-268. Cai, J., et al., N-methyl-N-nitro- N'-nitrosoguanidine induces the expression of CCR2 in human gastric epithelial cells promoting CCL2-mediated migration. Mol Med Rep, 2016. 13(2):p. 1083-90.), also unable to observe its long-term induction results

However, there is no report on the long-term induction of cell models by Helicobacter pylori and MNNG at home and abroad.

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