重组人p53腺病毒感染对携带不同p53状态胃癌细胞增殖和凋亡的影响

目的研究重组人p53腺病毒感染不同p53状态胃癌细胞对其p53蛋白表达、生长抑制率、细胞周期与凋亡率的影响。方法不同浓度重组人p53腺病毒感染3种不同p53状态胃癌细胞,即含野生型p53基因的细胞(wild-type)、含突变型p53基因的细胞(mutant-type)、含空载质粒即p53基因缺失的细胞(vector-cell)。48 h后,用Western blotting法检测p53蛋白在3种胃癌细胞中的表达;用MTT法测定重组人p53腺病毒感染3种胃癌细胞的生长抑制率,用流式细胞仪检测细胞周期分布和凋亡率。结果rAd-p53感染3种胃癌细胞48 h后p53蛋白表达阳性,对照组p53基因缺失的胃癌细胞无表达,对照组含野生型p53基因的细胞和含突变型p53基因的细胞弱表达。rAd-p53对3种胃癌细胞的生长抑制效应在一定的浓度范围内呈剂量依赖性,而与细胞内在的p53状态无关。含野生型p53基因的细胞、含突变型p53基因的细胞和p53基因缺失的细胞感染rAd-p53后诱导G2/M期阻滞与细胞凋亡率分别增加2.5、3.6、3.2倍。结论腺病毒介导p53基因感染3种不同p53状态胃癌细胞改变细胞内在的p53状态,p53蛋白表达、生长抑制率、细胞周期分布、凋亡率均与细胞内在的p53状态无关。

The effect of recombinant adenovirus p53 transfection on proliferation and apoptosis of gastric cancer cells with different types of p53

Objective To Study the effect of recombinant adenovirus p53 transfection on protein expression,growth inhibition rate,cell cycles and apotosis rate cells with different types of p53.Methods the recombinant adenovivus p53 in different and apotosis concentrations were infected.BGC-823 with three different p53 status: BGC-823 cells with wild-type p53 gene,BGC-823 cells with mutant type p53 gene and BGC-823 cells with p53 gene deletion were transfected with recombinant human p53 adenovirus.After 48 h,the expression of p53 protein in three gastric cancer cell lines was detected by Western blotting method;cell proliferation was determined by MTT,cell-cycle distributions and apoptosis rates were detected by Flow Cytometry.Results Three types of gastric cancer cells transfected with rAd-p53 expressed p53 protein.However,in the control groups,cells with wild type p53 and cells with mutant p53 express P53 weakly,cells with p53 gene deletion did not express p53 protein.Proliferation of three gastric cancer cell lines were rAd-p53 dose-dependent,regardless of the cell-endogenous p53 status.Induced G2/M phase arrests and apoptosis rates of cells with wild type p53,cells with mutant p53 and cells with p53 gene deletion transfected with rAd-p53 increased 2.5,3.6,3.2 times than controll groups,individually.Conclusion Transfected with adenovirus-mediated p53 gene,three different p53 types gastric cancer cells changed the p53 status.P53 protein expression,growth inhibitions rate,apoptosis rates and cell cycles were independent with cell-endogenous p53.