Effect Of Helicobacter Pylori On The Expression Of GATA3,Cx32、Cx43in GES-1Cells And The Correlation Analysis

objective:Our previous studies have found that the expression of Cx32, Cx43gradually reduced while GATA3increased in the H.pylori-infected gastric carcinogenesis at different stages which were called "inflammation-carcinoma chain"(chronic non-atrophic gastritis, chronic atrophic gastritis, intestinal metaplasia, dysplasia and gastric cancer). Based on the bioinformatics analysis that there were GATA3binding sites in the promoter regions of Cx32and Cx43genes by, the potential regulation of Cx32, Cx43by GATA3and its mechanism were of great interest. We established GES-1cell model to observe the effect of H.pylori on GATA3and the relationship between GATA3and Cx32, Cx43, and to explore whether H.pylori infection resulted in the decrease Cx32, Cx43through alternative expression of GATA3, which correlated with the gastric carcinogenesis as well.Methods:The H.pylori strains, isolated from gastric cancer patient, were co-cultured with GES-1cells for12hours and24hours. GES-1cells without H.pylori were corresponding control group. Real-time PCR and Western-blot were used to detect the expression of GATA3, Cx32, and Cx43in GES-1cells. Scrape-loading dye transfer technique was used to test gap junctional intercellular communication function in GES-1cells. At the same time, we transferred GATA3siRNA into BGC803by cationic liposome (the expression of GATA3was highest in BGC803among the gastric cancer cell lines in our pre-experiments)., then the expression of Cx32, Cx43in BGC803cells were detected on condition of successful GAT A3knock down.Results:(1) The expression of GATA3in GES-1cells in the experiment group was time-coursly increased both in transcription and protein expression (P<0.05).(2) The expression of Cx32, Cx43in H.pylori infected GES-1cells was time-coursly decreased in transcription and protein expression (P<0.05).(3)Change of the GJIC function:The control group showed better GJIC function in the GES-1cells, and the fluorescent dye migrated to3-4rows in the adjacent cells. Compared with the control group, the GJIC function of GES-1cells in the H.pylori infected GES-1cells showed weak GJIC function or no GJIC function, and most of the fluorescent dye was confined to the area of scratched single row cells and only a few migrated to1-2rows.(4) The expression of GATA3and Cx32, Cx43in transcription and protein (P<0.05) levels were negatively correlated after H.pylori infection. The expression of Cx32, Cx43in BGC803cells could be negatively regulated by GATA3both in transcription (P<0.05) and protein levels (P<0.05) due to GATA3siRNA experiments.Conclusions:(1).H.pylori triggers the expression of GATA3and disruption of Cx32, Cx43, and breaks the GJIC function in GES-1cells.(.2).The expression of GATA3is negatively correlated with Cx32and Cx43. GATA3negatively regulates expression of Cx32, Cx43and potentially play an important role in H.pylori-infected gastric carcinogenesis.