The Role Of Phospholipid Binding Protein Annexin A6 In Autophagy Of Gastric Cancer Cells

Background:The incidence and mortality of gastric cancer remain high worldwide especially in China,which seriously threatens human health.Gastric cancer cells are exposed to various stresses,including nutrition deprivation,endoplasmic reticulum stress and oxidative stress during the development and treatment.In this process,autophagy coordinates with stress response to maintain cell survival or promote cell death.Since the role of autophagy in tumors is double-sided,the molecular function of autophagy in different tumors needs further investigation and this may help us explain the role of autophagy in tumors more exactly.Objective:Our purpose is to investigate the regulatory effect and its mechanism of phospholipid binding protein Annexin A6 in autophagy of gastric cancer cells and to explore its effect on cell survival while exposing to stress.Methods:First of all,western blot was used to detected expression of Annexin A6 in different cell lines.The appropriate gastric cancer cell lines were selected to construct stable and inducible Annexin A6 overexpressed cell lines.The changes of autophagic flow were detected by western blot and immunofluorescence after Annexin A6 knockdown or overexpression and it was confirmed by treatment with or without autophagy inhibitor chloroquine followed by western blot.Western blot was then used to explore the mechanism of regulating autophagy.The effect of Annexin A6 on gastric cancer cells and its effect on cell viability while exposing to low glucose stress were verified by MTS cell viability assay and colony formation assay.Results:Annexin A6 inhibits autophagy through activating mTORC1 pathway and inhibits gastric cancer cell proliferation.Knockdown of Annexin A6-induced autophagy is beneficial for gastric cancer cells to survive under low glucose conditions caused by glycolysis inhibitor.