[Inhibitory effect of combined transfection of p53 and AS genes on K562 cell proliferation].

ABSTRACT

This study was aimed to investigate the inhibitory effect of combined transfection of p53 and angiostatin (AS) genes on K562 cells and to explore its mechanism. pVTRIO2-hp53-hAS was transfected into K562 cells with lipofectamine 2000, RT-PCR was used to determine the expression of gene of interest in transfected cells, MTT growth curve and flow cytometry were used to analyze the cell cycle for observation biological changes of cells, the cellular immunochemistry assay was used to observe the expression differences between VEGF, Bcl-2 and Bax proteins. The results indicated that the genes of interest have been transfected and stably expressed, the increase of K562 with genes of interest was slower than that without genes of interest (p<0.05). And the increase of K562 in double gene group was slower than that in p53 and AS groups (0.264+/-0.011 at last day A290 nm; 0.652+/-0.039 at last day A290 nm; 0.604+/-0.017 at last day A290 nm respectively) (p<0.05). After transfection, the expressions of VEGF and Bcl-2 protein decreased, but the expressions of Bax increased. It is concluded that the combined transfection of p53 and AS genes into K562 cells shows more notable and powerful inhibition on proliferation than those transfected with single one gene. The synergistic mechanism of p53 and AS genes may be commonly influenced the pathway of Bcl-2 and Bax expression.