Effects Of Luteolin On Gastric Cancer AGS Cell Line Overexpressing EGFR Or EGFRvⅢ

Objectives:The purpose of this study was to Establish gastric cancer AGS cell lines overexpressing EGFR and EGFRvⅢ and observe effects of luteolin on gastric cancer AGS cell line overexpressing EGFR or EGFRvⅢ.Methods:In this study,The EGFRvⅢ overexpression lentiviral vector was established.Human gastric cancer AGS cells cultured in vitro were respectively infected with overexpressing EGFR or EGFRvⅢ lentivirus,and the infection rate was calculated.Stable EGFR or EGFRvⅢ expressing gastric cancer cell lines were established.EGFR overexpressing group,EGFRvⅢ overexpressing group,negative lentivirus infection group and blank control group were setup.The expression of EGFR or EGFRvⅢ were dected by Western Blot.Results:1.PCR electrophoresis result showed that c DNA clone of EGFRvⅢ gene was successfully obtained.Sequencing results showed that the lentiviral vector plasmid was constructed correctly.The fluorescence result pointed out that the fluorescent gene was expressed normally.Real time quantitative PCR showed that EGFRvⅢgene was stably overexpressed in 293 T cells,and the EGFRvⅢ gene overexpression lentivirus was successfully constructed.2.After infection,the infection rate of lentivirus was more than 85% under fluorescence microscope.Western blot result shows that EGFR or EGFRvⅢ was overexpressed in two group.Human gastric cancer AGS cell lines overexpressing EGFR or EGFRvⅢ was constructed successfully.3.Compared with the blank control group and the negative control group,the overexpression EGFR group and the overexpression EGFRv Ⅲ group have significantly enhanced cell proliferation,migration and invasion.4.Luteolin inhibited cell proliferation in uninfected virus group,EGFR overexpression group and EGFRvⅢ overexpression group in a dose-and time-dependent manner.5.Luteolin can inhibit the migration and invasion of cells in each group,the difference was statistically significant.Conclusions:1.The constructed lentivirus can infect human gastric cancer AGS cell lines,and the EGFR gene and EGFRvⅢ gene can be stably overexpressed in human gastric cancer AGS cells.2.Overexpression of EGFR and EGFRvⅢ genes can significantly change the tumor biological characteristics of human gastric cancer AGS cells,making them have stronger proliferation,migration and invasion capabilities.3.Luteolin inhibits the biological activity of AGS cells,AGS cells overexpressing EGFR,and AGS cells overexpressing EGFRvⅢ to varying degrees.