摘要
背景:生存素(survivin)是凋亡抑制蛋白(IAP)基因家族成员之一,在多数肿瘤组织中高表达。目的:观察生存素反义寡脱氧核苷酸(ASODN)对人胃癌细胞株凋亡的影响。方法:将体外培养的人胃癌细胞株SGC-7901分为不同浓度生存素ASODN组、无关寡脱氧核苷酸(N-ODN)组和对照组。以四甲基偶氮唑蓝(MTT)试验检测生存素ASODN对SGC-7901细胞生长的影响;通过形态学观察、DNA电泳和流式细胞仪分析反映细胞凋亡情况;以端粒重复序列扩增-酶联免疫吸附测定(TRAP-ELISA)方法检测端粒酶活性。结果:生存素ASODN能抑制SGC-7901细胞生长,诱导细胞凋亡,抑制端粒酶活性。凋亡细胞形态学上表现为细胞膜起泡、染色质固缩、核碎裂、凋亡小体形成;DNA电泳呈现凋亡特征性阶梯状条带;流式细胞仪分析显示G1期前出现亚二倍体凋亡峰。结论:生存素ASODN能诱导人胃癌细胞株SGC-7901凋亡,抑制细胞生长及其端粒酶活性。
Background： Survivin, a member of the inhibitors of apoptosis protein （IAP） gene family, is highly expressed in a variety of cancers. Aims： To investigate the effect of antisense oligodeoxynucleotide （ASODN） targeting survivin on apoptosis of human gastric cancer cell line. Methods： Cultured human gastric cancer cell line SGC-7901 were allocated into control group, non-sense oligodeoxynucleotide （N-ODN） group, and survivin ASODN groups at different concentrations. Effect of survivin ASODN on the proliferation of SGC-7901 cells was examined by methyl thiazolyl tetrazolium （MTT） assay. The apoptotic cells were determined by morphological studies, DNA electrophoresis and flow cytometry analysis. The activity of telomerase was detected by telomeric repeat amplification protocol-enzyme-linked immunosorbent assay （TRAP-ELISA）. Results： After incubation with survivin ASODN in vitro, the proliferation of SGC-7901 cells was inhibited, the apoptotic rate increased significantly, and the telomerase activity decreased. Typical characteristics of apoptotic cells as membrane blebbing, chromatin pyknosis, karyorrhexis, and apoptotic body were seen under electromicroscope. Apoptosis-specific DNA ladder bands from SGC-7901 cells were visualized with electrophoresis. Hypodiploid apoptotic peak was observed before GI phase by flow cytometry analysis. Conclusions： Survivin ASODN may play a crucial role in the apoptosis of gastric cancer cell line SGC-7901, and inhibits the proliferation and telomerase activity of SGC-7901 cells.
出处
《胃肠病学》
2007年第8期477-480,共4页
Chinese Journal of Gastroenterology
