Effect Of Overexpression Of CHD1 On Tumor Biological Activity And Chemosensitivity Of AGS Gastric Cell Line

ObjectivesThe purpose of this study was to investigate the effect of CDH1 on tumor biological behaviors such as proliferation,migration,invasion,apoptosis and cell cycle of gastric cancer AGS cell line were observed and the overexpression of CDH1 on chemosensitivity of the cell line was detected.MethodsThe CDH1 overexpressed lentivirus vector was established to infect gastric cancer AGS cells cultured in vitro.The transfection rate was calculated by fluorescence count,and a stable CDH1 overexpressed gastric cancer cell line was established,and positive lentivirus transfection group,negative lentivirus infection group and negative control group were set up.The expression of cdh1 gene mRNA was detected by real-time PCR and the expression of E-cadherin was detected by Western blot.The effect of CDH1 overexpression on cell cycle and apoptosis of gastric cancer cells was detected by flow cytometry.Transwell model was constructed and cell scratch test was used to observe the effect of CDH1 overexpression on gastric cancer cell migration.The effects of overexpression of CDH1 on the proliferation of gastric cancer cells and the chemosensitivity of various chemotherapeutic drugs for gastric cancer were analyzed by CCK8 assay.Results1.Electrophoretic analysis of PCR products showed that cDNA clone of CDH1 gene was successfully obtained.Sequencing results showed that lentivirus transfection plasmid was constructed correctly.The fluorescence detection showed that the 293 T cells produced the lentivirus particles,and the Western blot showed that the CDH1 was expressed steadily in the cells,and the CDH1 gene lentivirus expression vector was successfully constructed.2.After CDH1 was transfected into AGS cells by lentivirus,the transfection rate was more than 80% by fluorescence microscope.Real-time PCR was used to identify the overexpression of CDH1 gene and Western blot to identify the high expression of CDH1 gene.3.The cell cycle,proliferation,migration,invasion and other biological activities of lentivirus transfected experimental group were significantly inhibited.The chemosensitivity of experimental group cells to 5-FU and PTX was increased.Conclusions1.Lentivirus can infect AGS cell line,and CDH1 gene can be stably overexpressed in AGS cells.2.After lentivirus-mediated CDH1 transfection into AGS cells,the biological activity of tumor cells is inhibited.It is further confirmed that CDH1 is a tumor suppressor gene.3.Overexpression of CDH1 in gastric cancer AGS cell line can enhance the chemosensitivity of gastric cancer cells to 5-FU and PTX,which makes it a new index to judge prognosis and guide drug use.