Lentivirus-mediated CD/TK gene selectively kills human gastric adneocarcinoma cells
AIM: To study the killing effect of lentivirus-mediated CD/TK double gene controlled by kinase insert domain-containing receptor (KDR) promoter on human gastric carcinoma cells SGC-7901 in vitro. METHODS: SGC-7901 cells (with KDR expression) were transfected with FGW-KDRP-CD/TK vector in v/tro. The infection efficiency in SGC-7901 cells was observed under fluorescence microscope. And the expression of CD/TK in genetically modified cells was detected by RT-PCR and Western Blot. The cell growth curve was drawn according to cell counts. Flow cytometer(FCM) was used for cell cycle analysis in the treatment group. Followed by treatment of the ganciclovir(GCV) and 5-fluorocytosine (5-FC), the killing and bystander effects of suicide gene therapy system on SGC-7901 cells were evaluated by MTT" method. RESULTS: The transfection efficiency in SGC-7901 cells increased with the increasing lentiviral titer. RT-PCR and Western Blot demonstrated that there existed the CD/TK gene and the protein gene product in genetically modified cells. No significant difference was found in the characteristics of growth between the transfected SGC-7901 cells and the non-transfected cells with lentiviral vector according to cell growth curve. Flow cytometry analysis showed the proportion of S phase cells decreased in the treatment group compared with the control group as increasing the prodrug concentration. Prodrug could inhibit proliferation of SGC-7901 cells and the effect was dose-dependent by MTY analysis. In addition, a considerable bystander effect was also observed in the suicide gene system. CONCLUSION: CD/TK fusion gene system driven by KDR promoter can selectively kill SGC-7901 cells, and there exists an obvious bystander effect during this process.
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