The Regulatory Effect And Mechanism Of LINC00152 In Helicobacter Pylori-infected Gastric Epithelial Cells

PURPOSES:The purpose of this study is to explore the regulatory role and mechanism of LINC00152 in H.pylori-infected gastric epithelial cells,and provide a molecular basis for the diagnosis of H.pylori infection-related gastritis and gastric cancer.METHODS:(1)qRT-PCR was performed to detect the expression of LINC00152 in H.pylori-infected gastric mucosal epithelial cells,and determine the relationship between LINC00152 gene expression and H.pylori.qRT-PCR was performed to detect LINC00152 in different gastric cancer cells and normal gastric mucosal epithelium.(2)The si-LINC00152 and the overexpressed plasmid pc DNA-LINC00152 were transfected into different gastric cancer cells respectively by gene interference technology and gene overexpression technology.The knockdown and overexpression cell models of LINC00152 were established in vitro,and the results were verified by qRT-PCR.(3)The effect of LINC00152 on the proliferation of gastric cancer cells was detected by CCK8 cell proliferation assay.The effect of LINC00152 on gastric cancer cells was detected by transwell cell migration assay.(4)Western blot was performed to detect the effect of LINC00152 on NF-ΚB and Wnt-β-catenin signaling pathway.(5)EGFP-LC3 plasmid was co-transfected into both low-expression and high-expression LINC00152 cells by cell transfection technique,and the aggregation of LC3 particles in cells was observed to analyze the difference of autophagy levels in each group.(6)Western blot was used to detect the effect of LINC00152 expression on the expression levels of autophagy related proteins in each group.RESULTS:(1)LINC00152 was expressed in normal gastric epithelial cells GES-1 and different gastric cancer cell lines,with the highest expression in BGC-823 cells and the lowest expression in MGC-803 cells.(2)The expression of LINC00152 was inhibited in GES-1 infected with H.pylori,and the expression level of LINC00152 showed a downward trend with the extension of H.pylori infection time.(3)Transwell cell migration assay and CCK8 cell proliferation assay indicated that LINC00152 expression could inhibit the proliferation and migration of gastric cancer cells induced by H.pylori infection.Meanwhile,the migration ability of gastric cancer cells was affected by the regulation of TIMP1,MMP9,MMP2 and other proteins.The proliferation ability of gastric cancer cells was affected by regulating PCNA,Bcl-2 and Cyclin D1 cell proliferation-related proteins.(4)LINC00152 affected the EMT process of gastric cancer cells by regulating mesenchymal cell marker proteins N-cadherin,Snail,Vimentin and epithelial cell marker protein E-cadherin.(5)LINC00152 regulated the changes of key proteins in NF-κB and Wnt-β-catenin signaling pathways.(6)The expression of LINC00152 could affect the autophagy of gastric epithelial cells,and H.pylori significantly promoted autophagy;Overexpression of LINC00152 could inhibit H.pylori induced autophagy in gastric cancer cells,while knockdown of LINC00152 can enhance autophagy.(7)The expression of LINC00152 can affect the expression of autophagy-related proteins LC3I/II,Beclin1 and P62.CONCLUSION:The expression of LINC00152 decreased in GES-1 infected with H.pylori,and affected the proliferation and metastasis of gastric cancer cells by regulating Wnt-β-catenin signaling pathway and EMT process,and also affect the autophagy and inflammation induced by H.pylori.