反义端粒酶RNA抑制肝癌细胞生长的机制

Effect of antisense telomerase RNA on SMMC7721

目的 观察反义端粒酶 RNA对 SMMC- 772 1肝癌细胞系的作用 ,探讨抗端粒酶治疗在原发性肝癌治疗中的意义 .方法 采用脂质体介导的方式将反义端粒酶 RNA真核表达载体 p BBS2 12 / h TR转入 SMMC772 1细胞中 ,经潮霉素筛选 ,克隆细胞株扩增鉴定后 ,采用 TRAP- PCR- EL ISA,FCM及电镜检测反义端粒酶 RNA对转染细胞端粒酶活性和细胞凋亡的影响 ;同时进行裸鼠荷瘤生长实验观察 .结果 TRAP- PCR- EL ISA法检测细胞端粒酶活性的结果为 :实验组吸光度值为 0 .980 ,对照组吸光度值为 2 .86 1;FCM检测细胞凋亡结果为 :实验组 13.8% ,对照组未见有凋亡峰形成 ,电镜观察发现转染细胞表现出典型的凋亡细胞形态 .裸鼠移植瘤生长实验发现 ,转染细胞生长明显受到抑制作用 ,说明反义端粒酶 RNA显著抑制 SMMC772 1细胞的端粒酶活性 ,并且对其有显著的促凋亡作用 .结论 反义端粒酶 RNA能有效地抑制肝癌细胞端粒酶活性 ,同时显著促进癌细胞凋亡 .

AIM To observe the effect of antisense telomerase R NA on SMMC7721 hepatocarcinoma cell line and to investigate the significance of anti-telomerase therapy for primary hepatoc arcinoma. METHODS pBBS212/hTR, antisense telomerase RNA gene ex pression vector, was intro-duced into SMMC7721 cells by using lipofectin mediation. After hygromycin screening and cloning, TRAP-PCR-ELISA a nd FCM were used to observe the effects of pBBS212/hTR on telomerase activity an d cell apoptosis. Then the animal model was established by transplanting cells (SMMC7721 cell strain transfected by pBBS212-hTR gene) into the abdomens of nude mice. RESULTS TRAP-PCR-ELISA showed tha t the experimental group registered 0.980 and the control group 2.861 in telomerase activity; FCM showed that the experimental group registered 13.8% in apoptosis, suggesting that antisense telomerase RNA had a significant inhibition of SMMC7721 telomerase activity and a marked promotion of cell apoptosis, and that in the meantime it could inhibit cell proliferation. CONCLUSION Inhibition of hepatocarcinoma telomerase activity and promotion of carcinoma cells' apoptosis through antisense telomerase RNA provide an approach to the treatment of hepatocarcinoma.