腺病毒介导的P53基因抗人胃腺癌增殖的研究

Anti-proliferative effect of type P53 gene mediated by adenovirus on human gastric carcinoma

目的 研究腺病毒介导的野生型P5 3基因 (Ad -P5 3 )抗人胃腺癌细胞增殖的效果。方法 利用携带P5 3基因的复制缺陷型腺病毒 (Ad -P5 3 )感染人胃腺癌细胞株SGC - 790 1,以携带报告基因LacZ的同型载体 (Ad -LacZ)作对照 ,MTT法测定及克隆形成实验分析P5 3基因对胃腺癌细胞的体外抑瘤效应。用流式细胞技术分析P5 3基因转染后对细胞周期的影响以及引起凋亡情况 ,并用电镜观察细胞的超微结构变化。结果 野生型P5 3基因转移对SGC - 790 1具有明显的抗增殖作用。Ad -P5 3感染SGC - 790 1后细胞生长明显被抑制 ,抑制率达 88 73 % (MOI为 2 0 0 ,7d)。克隆形成能力也明显受到抑制 ,当MOI分别为 5 0 ,10 0 ,2 0 0时 ,克隆形成数为 2 2 ,0 ,0。流式细胞计数分析显示Ad -P5 3能引起SGC - 790 1细胞发生周期阻滞和细胞凋亡 ,凋亡百分率达 71 4% (MOI为 2 0 0 ,7d)。电镜观察结果也显示细胞出现凋亡的特征性形态改变。结论 腺病毒成功介导野生型P5 3基因转移入人胃腺癌SGC - 790 1细胞中 ,野生型P5 3基因有显著抑制胃腺癌细胞增殖的作用。

Objective To evaluate the effect of the anti-proliferative effect of wild type P53 gene transfection mediated by recombinant adenovirus (Ad-P53) on human gastric carcinoma.Methods To analyze the anti-tumor effects of wild type P53 gene in vitro using the proliferation-defective recombinant adenovirus as the vector.The SGC-7901 cells,derived from human gastric carcinoma,were infected with the proliferatively defective recombinant adenovirus bearing wild type P53 gene (Ad-P53) or LacZ gene (Ad-LacZ),the latter considered as the vector control.The effects of P53 gene on cell proliferation was evaluated by using MTT staining method or colony performing experiment.The mechanism of the anti-proliferation was analyzed under the electro-microscopy and flow cytometry (FCM).Results The transfection of wild type P53 gene into SGC-7901 cells showed anti-proliferation effects.The cell growth was inhibited by Ad-P53 infection with inhibition rate of 88.73% (MOI was 200,day 7).And the colony performing ability of the tumor cell was significantly inhibited as well,the numbers of the colonies were 22,0 and 0,while the cells were infected with the virus of 50,100 and 200 MOI,respectively.The SGC-7901 cells infected with Ad- P53 analyzed by FCM showed the impeding of cell growths and apoptosis.The percentage of apoptosis was 71.4% (MOI was 200,day 7).Conclusions Wild type P53 gene was transferred into SGC-7901 cells and showed significant anti-proliferative effects on the cell growth.The expression of the P53 gene can cause the impeding of cell growth and apoptosis.