The Inhibition Of Combined Transfection Of P53 And AS Genes Into K562

Recent studies show gene therapy has remarkable advantage compared to thetraditional methods. Studies demonstrate most of malignant tumor cells includingleukemia ceil line K562 express VEGF and its receptor. VEGF can promoteangiogenesis and proliferation of tumor. So it will inhibit angiogenesis and growth oftumor by suppressing VEGF. Angiostatin (AS) can efficiently suppress the expressionof VEGF. P53 is a tumor suppressor gene. They have different mechanism in the fieldof inhibit tumors. OBJECTION Our study is about to observe whether p53 and AShave synergic depression in K562 or not and expire its mechanism by co-transfectionp53 and AS genes. METHOD Lipofectamine2000 is used to transfectp53,AS,pVTRIO2-p53-AS into K562. 16 days later, expression of target genes isdetected with RT-PCR. MTT growth curve, FCM are used to analyze the results. Theexpression of VEGF, bcl-2 and bax is tested with cell immunochemistry. RESULTRT-PCR shows the target genes have been tranfected successfully and expressedstably. MTT shows the rise rate of K562 with target genes is lower than that of thegruop without transfection(p＜0.05). And the co-transfected group (0.264±0.011, lastday's A_290nm)iS slower than p53 group(0.652±0.039, last day's A_290nm)and ASgroup(0.604±0.017, last day's A_290nm)(p＜0.05). Compared with the controlledgroup(37.68±2.10)%, data of FCM show cell's percentage in G₁ phase of p53group(43.14±2.00) % and co-transfected group(42.28±1.98)% show statisticdifference (p＜0.05), but the AS group(35.56±1.98)% doesn't (p＞0.05). Aftertransfection, the expression VEGF and bcl-2 deceases; nevertheless bax increases.CONCLUSION The study shows that the depression of combined transfection of p53and AS on K562 is more notable and powerful than either single one. P53 and ASmay co-inhibit VEGF, influent the rate of bcl-2/bax and induce the cell's apoptosisfinally.