STABLE CELL STRAINS EXPRESSING LOWER bcl-2 GENE WERE ESTABLISED VIA bcl-2 TARGETING SHORT HAIRPIN RNA PLASMID VECTOR

Objective To construct a short, bcl-2 gene-targeting hairpin RNA (shRNA) plasmid vector, which was transfected to high-bcl-2-gene-expressed gastric cancer line SGC-7901, a cell strain with stable and low bcl-2 gene expression was then screened.MethodsFour pairs of oligonucleotide specific for bcl-2 gene were designed, synthesized, and inserted intopGPH1/GFP/Neo plasmids. The vectors were transfected into SGC-7901 cells with lipofectamine. RT-PCR was used to detect bcl-2 gene expression and MTT technique employed to detect the proliferation of tumor cells after bcl-2 gene was silenced. Cells with the strongest inhibition were selected with G418 and the stable cell clones obtained.ResultsAs compared with the control, themRNA expression of bcl-2 in the transfected cells was significantly suppressed, the rate of cell proliferation was significantly reduced(t=2.41,P0.05). ConclusionSGC-7901 cell strains, which express stable and lower level of bcl-2 mRNA, were successfully obtained by using bcl-2-targeting shRNA plasmid vector. The present study provided basis for research on gene therapy of gastric cancer.