The Function And Mechanism Of LAPTM4B In Gastric Carcinogenesis And Progression

ObjectiveThe present study was to demonstrate the expression of LAPTM4B that regulated EMT-related proteins and EGFR in Helicobacter pylori infection in cell line GES-1.The research investigated the function and mechanism of LAPTM4B in H.pylori-related gastric cancer,which resulted in proliferation disorder and enhancing invasion capability obviously,the prevention and therapy may be provided.MethodsThe study was consisted of two parts.In the first part of the experiment,the expression and significance of LAPTM4B-35,EMT-related proteins(ZO-1,ZEB1,E-cadherin,?-catenin,Vimentin,Snail,Slug)and EGFR during the process of H.pylori infection were inspected validly by means of Western blot.Firstly,the Human Gastric Epithelial Cell line(GES-1)and H.pylori 26695 were co-cultured in vitro,subsequently,the cells were collected respectively at corresponding time points(Oh,12h,24h and 48h),the expression of LAPTM4B-35,ZO-1,ZEB1,E-cadherin,?-catenin,Vimentin,Snail,Slug and EGFR were investigated by Western blot.Based on experimental method of plasmid transfection to overexpress the LAPTM4B,studying the expression of EMT-related proteins and EGFR were regulated by LAPTM4B.Further,to research the expression EMT-related proteins and EGFR caused by aberrant LAPTM4B gene in H.pylori infection,by employing the method of siRNA to silence the LAPTM4B before cocultivation in vitro,the proteins were inspected and observed with uniformly experimental approach.In the second part of the research,we study the relationship among the LAPTM4B,EMT-associated proteins as well as the protein of EGFR.In addition,the research of the anterior protein had effects on cell proliferation and cell invasion ability in gastric cancer cell line SGC-7901.First of all,the different cell lines expression level of LAPTM4B-35 were detected and the cell of high expression was selected to down-regulated the expression of LAPTM4B.After the successful transfection and identification,the expression level of the EMT-related proteins(ZO-1,ZEB1,E-cadherin,?-catenin,Vimentin,Snail,Slug)and EGFR were surveyed by means of Western blot.Subsequently,the proliferation of cell was dectected by the method of CCK-8.The changes of ability in invasion and capacity in proliferation were tested by Transwell invasion experiment testing.ResultsThe frist experimental study mainly revealed that the expression level of LAPTM4B-35 was low in GES-1,however,after co-culturing the GES-1 with H.pylori,LAPTM4B was increased significantly.Surprisingly,the expression of the EMT-related proteins and EGFR were markedly different,the proteins of ZEB1,?-catenin,Vimentin,Snail,Slug were up-regulated expression,ZO-1,E-cadherin were signally down-regulated expression.EGFR were up-regulated expression.Besides,based on experimental method of plasmid transfection to overexpress the LAPTM4B,the study presented the identical result.The proteins of ZEB1,?-catenin,Vimentin,Snail,Slug were up-regulated expression,while the ZO-1,E-cadherin were down-regulated expression in GES-1.EGFR was down-regulated expression.The cell proliferation was promoted by the CCK8 experiment.The mobility capacity of cells was increased with the help of transwell chamber.With an in-depth study,the LAPTM4B gene was down-regulated by gene knock down in GES-1 and co-cultured with H.pylori,observing the identical changes of EMT-associated proteins and EGFR.The second experimental survey uncovered the expression of LAPTM4B-35 in SGC-7901 that expressed higher than GES-1.The RNAi technology was used to effectually silenced the LAPTM4B in SGC-7901 and examined the EMT-related proteins and EGFR,discovering that the proteins of ZEB1,?-catenin,Vimentin,Snail,Slug were down-regulated expression evidently,nevertheless,E-cadherin,ZO-1 were overexpressed.EGFR was down-regulated expression.Beyond that,the proliferation ability of cell was deciline by the method of CCK-8.The invasion ability was decreased by transwell chamber.Conclusions1.The regulation of LAPTM4B affected the expressions and functions of EMT-related proteins and EGFR,resulting in the H.pylori-related gastric cancer.2.The LAPTM4B gene was involved in inducing the expression of EMT-related proteins and EGFR,resulting in enhanced capacity of proliferation and invasion in gastric carcinogenesis and progression.