Analysis Of The Alternative Splicing Regulation In Helicobacter Pylori-induced Gastric Epithelial Carcinogensis

Analysis of the alternative splicing regulation in Helicobacter pylori-induced gastric epithelial carcinogensisObjectiveWarran and Mashall since 1983 has been found in Helicobacter pylori, people and animal experiments were Table H. pylori infection and gastric cancer is closely related.1994, IARC (IRAC/WHO) to H. pylori in human gastric cancer specific for the class I carcinogen. The formation of gastric cancer is a multi-factor, multi-step complex process. H. pylori infection, host genetic factors and environmental factors determine this process. But so far, H. pylori carcinogenic mechanism is still not exactly clear. Most in vitro studies showed that chronic gastritis-atrophic gastritis-intestinal metaplasia dysplasia-gastric such a long process, H. pylori may act on the early stages of gastric cancer. H. pylori can occur in many ways led to a series of damage in epithelial cells, gastric cancer and genetic damage, mutation, changes in intracellular signal transduction and cell proliferation, apoptosis, the loss of balance and so on. This article is a regulation of H. pylori infection in gastric cancer related gene transcription and splicing mechanisms for exploratory research, a system transcriptomic study from concept to actual important attempt. The perspective of the alternative splicing H. pylori infection causes gastric cancer, the molecular mechanism of pathogenesis of gastric cancer will have a better understanding of, and for drug development to develop new ideas.Methods 18 patients randomly selected patient diagnosed asH. pylori-positive gastric cancer which, taken at 5 cm adjacent biopsy specimens were isolated and cultured H. pylori.Use the liquid medium for H. pylori bacteria in shaking to obtain H. pylori culture filtrate. cagA gene positive H. pylori culture filtrate into gastric epithelial cell line GES-1 cells. On cell morphology, growth, colony formation ability of clone detection identification of transformed cells model.Extraction of RNA extraction kit with GES-1 cells and H. pylori culture filtrate into the GES-1 cell total RNA. Using gene specific primers by nested reverse transcription PCR, combined with identification of CD44 splicing variants.ResultsThe experiment successfully from the Armed Police General Hospital endoscopy room collected gastric biopsy specimens, isolated and cultured cagA gene positive H. pylori. Use of liquid medium on H. pylori bacteria in shaking to obtain H. pylori culture filtrate. CagA gene positive H. pylori established culture filtrate into gastric epithelial cell line GES-1 cell model. Using gene specific primers by nested reverse transcription PCR, combined with identification of the GES-1 cells and H. pylori culture filtrate into the GES-1 cells in CD44 splicing variants.Of the cagA gene positive H. pylori culture filtrate into gastric epithelial cell line GES-1 cell model checking:(1) morphological changes:the model cell morphology appeared pleomorphic, irregular nuclear morphology, cells were not seen this change.(2) cell growth curve:With H. pylori culture filtrate concentration and treatment time, the growth curve was increased gradually, and a dose-time is relevant.(3) Soft agar colony growth:a different titer H. pylori culture filtrate GES-1 cells, cloning efficiency of the control group 30%,5%(v/v) H. pylori filtrate treated cloning efficiency 45%,10%(v/v) H. pylori filtrate treated cloning efficiency 55%,5%(v/v) H. pylori filtrate treated cloning efficiency compared with the control group were significantly different (P<0.05),10%(v/v) H. pylori filtrate cloning efficiency and the treatment group compared with control group was significantly higher (P<0.01). Clones to form colonies in the size and number with H. pylori culture filtrate concentration increases.ConclusionPatients with gastric cancer from the patient isolated and cultured tissues cagA gene positive H. pylori. The method uses the light Leunk liquid medium for H. pylori bacteria in shaking to get the H. pylori culture filtrate. CagA gene positive H. established pylori culture filtrate into gastric epithelial cell line GES-1 cell model. Using gene specific primers by nested reverse transcription PCR, combined with identification of the CD44 splicing variants. Band of the experimental group and control group differences. This shows that H. pylori in gastric epithelial cell line transformed GES-1 cells may have occurred during the differential expression of CD44 gene, we hypothesized that cagA gene positive H. pylori in gastric cancer caused by aberrant splicing event is H. pylori induced an important molecular mechanism of gastric cancer. Our view from the alternative splicing of H. pylori induced gastric cancer mechanism for future research provides a new way of thinking.