人端粒酶反义寡脱氧核苷酸对胃癌细胞系SGC-7901的生长抑制和诱导凋亡的研究  

Effect of Human Telomerase Antisense Oligodeoxyribonucleotides on the Inhibition of Growth and Induction of Apoptosis in Gastric Cancer Cell Line SGC-7901

作　　者：叶静[1] 吴云林[1] 乔敏敏[1] 郭立霞[2] 谢弘[2] 

机构地区：[1]上海第二医科大学附属瑞金医院消化内科,200025 [2]中科院上海生命科学院细胞与生物化学研究所

出　　处：《胃肠病学》2003年第4期195-199,共5页Chinese Journal of Gastroenterology

基　　金：卫生部内科消化重点实验室开放基金(WKL200010)

摘　　要：端粒酶活性在胃癌组织中表达率很高,在正常胃黏膜组织中则基本不表达。因此,以端粒酶为靶点的反义治疗有望成为胃癌治疗的有效途径之一。目的:检测特定的人端粒酶反义寡脱氧核苷酸(AS-ODN)对胃癌细胞系SGC-7901生长的抑制作用,并证明其主要机制为抑制端粒酶活性和诱导细胞凋亡。方法:用改良的聚合酶链反应(PCR)-端粒重复扩增协议(TRAP)法定量检测端粒酶AS-ODN作用前后SGC-7901细胞的端粒酶活性;台盼蓝染色法观察SGC-7901细胞活力;光镜和电镜观察凋亡细胞形态,流式细胞仪检测细胞凋亡。结果:SGC-7901细胞有端粒酶活性表达。不同浓度的端粒酶AS-ODN作用于SGC-7901细胞48～96h后,细胞端粒酶活性和生长受到明显抑制,光镜、电镜观察和流式细胞仪检测均证实有细胞凋亡存在。错义AS-ODN(N-ODN)处理组SGC-7901细胞则无显著变化(P<0.05)。结论:端粒酶AS-ODN通过抑制胃癌细胞的端粒酶活性和诱导细胞凋亡,最终抑制胃癌细胞生长,其抑制作用呈剂量、时间依赖性和序列特异性。端粒酶AS-ODN对端粒酶活性的抑制比对细胞生长的抑制更敏感,两种抑制作用并不完全平行。Telomerase activity is highly expressed in gastric cancerous tissue, while almost no telomerase activity can be found in normal gastric mucosa. Hence, antisense therapy on telomerase may be a promising and effective measure for gastric cancer. Aims: To investigate the inhibitory effect of specialized human telomerase antisense oligodeoxyribonucleotides (AS-ODN) on the growth of gastric cancer cell line SGC-7901 and to verify the main mechanisms of telomerase AS-ODN on SGC-7901 cells in inhibiting telomerase activity and inducing cell apoptosis. Methods: Modified polymerase chain reaction (PCR)-telomere repeat amplification protocol (TRAP) was used to quantify the telomerase activity of SGC-7901 cells before and after treatment with telomerase AS-ODN. The growth and activity of SGC-7901 cells were explored using trypan blue staining. The morphology of apoptotic cells was observed by light and electron microscopy, and flow cytometry was used to detect the cell apoptosis. Results: Telomerase activity could be detected in SGC-7901 cells. After treatment with telomerase AS-ODN of various concentrations for 48～96 hours, telomerase activity and growth inhibition were found in SGC-7901 cells. Light and electron microscopic examination and flow cytometry found the presence of apoptotic cells. However, there was no significant effect on SGC-7901 cells after treatmen twith telomerase non-AS-ODN (N-ODN) (P<0.05). Conclusions: Telomerase AS-ODN can inhibit the growth of gastric cancer cells by telomerase activity inhibition and induction of cell apoptosis, which depend on the dosage, time and sequence specificity. The inhibitory effect of telomerase AS-ODN on the telomerase activity is more sensitive than that on the cell growth, and the two effects are not paralleled.

关 键 词：人端粒酶反义寡脱氧核苷酸 胃癌 细胞系 SGC-7901 生长抑制 诱导凋亡 

分 类 号：R735.2[医药卫生—肿瘤]