Construction Of Stable Over-expressing CD14 Gastric Adenocarcinoma SGC-7901 Cell System

Objective:A lentiviral vector capable of stably overexpressing CD14 was constructed.A gastric adenocarcinoma SGC-7901 cell line that stably overexpressed CD14 was established to investigate the expression changes of CD14 mRNA and protein after transfection of lentiviral vectors into SGC-7901 cells,laying the foundation for the study of the pathogenesis of gastric cancer.Methods:Construction of a CD14 over-expression lentiviral vector.Grouping: blank control group(SGC-7901 group),negative control group(NC-LV group),CD14 overexpression group(LV-CD14 group),NC-LV group transfected with CD14-free lentiviral vector SGC-7901 cells and LV-CD14 groups were transfected with SGC-7901 cells with CD14 overexpression vector.The blank control group received no treatment.Subsequently,mRNA and protein expression was detected by Real-time PCR and Western blot to investigate the effect of CD14 overexpression on total protein expression.Results: A lentiviral vector overexpressing CD14 was successfully constructed.Gastric adenocarcinoma SGC-7901 was successfully transfected with lentiviral vector over-expressing CD14,and a gastric adenocarcinoma SGC-7901 system with stable over-expression of CD14 was selected.The relative expression of CD14 mRNA in the LV-CD14 group was 4.32 times that of the NC-LV group and 4.82 times that of the blank control group.The relative expression level of the LV-CD14 group was statistically different from that of the NC-LV group and the blank control group.Significance(P<0.05).There was no significant difference between NC-LV group and SGC-7901 group(P>0.05).The protein expression level of the LV-CD14 group was 1.700 times that of the NC-LV group and 1.7419 times that of the blank control group.The cells that stably over-expressed CD14 had increased mRNA and protein expression levels,which was statistically significant compared with NC-LV group and blank control group(P<0.05).Conclusion: The gastric cancer SGC-7901 cells were successfully transfected with the CD14 overexpression lentiviral vector.In this study,CD14 mRNA and protein were over-expressed in gastric cancer cells after transfection with CD14 over-expression lentiviral vector.