重组人p53腺病毒联合奥沙利铂对胃癌细胞SGC-7901的生长抑制作用

Effects of Recombinant Adenovirus-p53 Combined with Oxaliplatin on Growth Inhibition of Human Gastric Cancer Cell Line SGC-7901

目的观察外源p53基因在胃癌细胞SGC-7901内的表达及其对胃癌SGC-7901细胞生长的影响、对胃癌细胞化疗敏感度的作用及机制。方法用重组人p53腺病毒注射液(rAd-p53)及奥沙利铂(OXA)单独及联合作用于胃癌细胞株SGC-7901不同时间后,CCT-8法检测其对体外培养SGC-7901细胞的抑制率,免疫组织化学SP法检测p53蛋白的表达情况,流式细胞仪(FCM)分析其细胞凋亡蛋白Caspase-3的表达情况。结果 rAd-p53及OXA单独作用SGC-7901细胞时,随药物浓度及作用时间的增加,细胞的生长抑制率逐渐增高;两者联合作用48h,其在较低浓度时即可显著抑制细胞生长(P<0.05);OXA(6.25μg/ml)与rAd-p53(5×107、5×108、5×109vp/ml)联合作用48h后,胃癌细胞caspase-3蛋白的含量较对照组升高,但p53蛋白无明显升高。结论 OXA和rAd-p53单药可抑制胃癌细胞的生长,两者联合对胃癌细胞的抑制作用明显增强;rAd-p53有增强OXA化疗敏感度的作用,其机制与通过线粒体途径激活下游的caspase-3诱导细胞凋亡有关。

Objective To observe the expression of exogenous p53 gene in gastric cancer cells and its effects on the growth of tumor cells;and to investigate the effects of adenovirusmediated p53 gene on chemosensitivity of human gastric cell and the value of gene therapy combined with chemotherapy.Methods Toinvestigate the effects of recombinant adenovirus-p53（rAd-p53） and oxaliplatin（OXA） alone and combined on the gastric cancer cell line SGC-7901 for different times,CCK-8 assay was used to examine the suppressive rate of cell growth dealt;p53 protein expression was detected by immunohistochemistry assay;and protein caspase-3 expression in cell was induced by different drugs alone and in combination by flow cytometry.Result When rAd-p53 and OXA was alone used to treat the gastric cancer cell line SGC-7901,with an increase in drug concentration and treated time,the cell growth inhibition rate gradually increased;when rAd-p53 and OXA was combined to treat the gastric cancer cell line SGC-7901 for 48 h,the cell growth inhibition rate gradually increased at the lowest concentration.When rAd-p53（5×107,5×108,5×109vp/ml） and OXA（6.25μg/ml） was combined to treat the gastric cancer cell line SGC-7901 for 48h,compared with the control group,the content of caspase-3 protein in gastric cancer cell gradually increased,but p53 protein expression didn＇t increase.Conclusion The proliferation of SGC-7901 could be inhibited by rAd-p53 OXA alone,but when rAd-p53 was combined with OXA,the proliferation of cells was higher than that used alone.The combination of rAd-p53 and OXA induced cell apoptosis through mitochondrial pathway to activate downstream of caspase-3.