摘要
目的探讨腺病毒载体介导的早幼粒细胞白血病基因(PML)生长抑制因子诱导胃癌细胞凋亡作用的机制。方法将人PML全长cDNA插入穿梭质粒pSGCMV,再与腺病毒骨架质粒pPE3共转染293细胞后获得重组病毒。用重组病毒感染胃癌细胞系SGC-7901,采用噻唑蓝比色法、流式细胞术以及免疫细胞化学法对p53和bcl-2在肿瘤细胞内的表达以及细胞凋亡的定性与定量指标进行检测。结果经腺病毒介导的PML处理的SGC-7901细胞生长受到明显抑制,凋亡细胞发生率升高,且与MOI值呈正相关,MOI值由5增加到20,细胞的生长抑制率从22．4％增加到38．5％,而细胞凋亡率从37．2％增加到49．8％。经腺病毒介导的PML处理的SGC-7901细胞内p53蛋白表达较对照组明显增加,bcl-2蛋白的表达则无明显变化。结论腺病毒介导的PML对胃癌细胞的杀伤主要是通过诱导细胞凋亡,p53蛋白高表达为其诱发胃癌细胞凋亡的机制之一。
Objective To investigate the apoptosis induced by adenovirus mediated PML growth suppressor in SGC-7901 gastric cancer ceils. Methods Human full-length PML cDNA was cloned into shuttle plasmid pSGCMV, the constructed plasmid containing PML gene was co-transfected into 293 cells together with backbone plasmid pPE3 to obtain recombinant adenovirus Ad-pml. Ad-pml was used to infect SGC-7901 cells. The expression of p53 and bcl-2 in SGC-7901 cells and qualitative, quantitative index of cell apoptosis were detected with MTT, flow cytometry and immunohistochemical method. Results The growth of SGC-7901 cells was inhibited and the apoptosis rate was increased by adenovirus mediated PML in a MOI- dependent manner. The expression of p53 protein was increased and that of bcl-2 protein was unchanged in SGC-7901 cells after treated with adenovirus mediated PML. Conclusions Adenovirus mediated PML exerts its cytotoxic effect on SGC-7901 ceils by inducing apoptosis. The increase of p53 protein expression is the mechanism inducing gastric cancer cell apoptosis.
出处
《中华普通外科杂志》
CSCD
北大核心
2006年第8期591-593,共3页
Chinese Journal of General Surgery
基金
黑龙江省科技攻关基金资助项目(编号G00C 1902)
关键词
胃肿瘤
基因表达
细胞凋亡
Stomach neoplasms
Gene expression
Apoptosis
