核酶切割技术对胆囊癌细胞端粒酶活性抑制作用的研究  

Ribozyme mediated inhibition on GBC-SD gallbladder carcinoma cells

作　　者：靳斌[1] 王伟[2] 姜希宏[1] 胡三元[1] 张朝阳[3] 乔筱玲[1] 

机构地区：[1]山东大学齐鲁医院肝脏移植中心,山东济南250012 [2]山东大学分子生物学实验中心,山东济南250012 [3]山东大学齐鲁医院普通外科,山东济南250012

出　　处：《中国普通外科杂志》2006年第3期198-201,共4页China Journal of General Surgery

基　　金：山东省医药卫生科研资助项目(21300005210431)

摘　　要：目的探讨锤头状核酶切割技术对胆囊癌细胞端粒酶活性的影响及对胆囊癌细胞生长增殖的作用。方法根据拟要切割的胆囊癌端粒酶RNA亚基模板两翼区的序列,体外合成锤头状核酶的基因序列,并构建入pTriEx-4真核表达载体。酶切及测序鉴定正确后,采用脂质转染法导入胆囊癌细胞,观察其对胆囊癌细胞端粒酶活性的影响及对细胞生长、增殖的作用,并设空载体转染组和单纯脂质体转染组作为对照。结果与对照组比较,实验组胆囊癌细胞的端粒酶活性明显减低;核酶基因作用后,G0/G1期细胞比例明显增高,S期细胞比例明显降低;细胞的分裂、增殖受到明显抑制。核酶作用后1 0 d凋亡率为1 7.1 0%,1 3 d后凋亡率为3 1.0 1%。结论核酶切割技术对胆囊癌细胞端粒酶活性有明显的抑制作用,并能抑制胆囊癌细胞的生长,促进细胞凋亡。Objective To study the effect of ribozyme mediated inhibition on telomerase activity and cell proliferation of GBC-SD cells. Methods According to the hTR template two wings region sequence, the hammerhead ribozyme gene sequence was synthesized in vitro. Then, it was engineered into the eukaryon expression vector pTriEx-4. Lipofectamine was used to transfect the carcinoma of gallbladder. The inhibition of telomerase activity and cell proliferation in GBC-SD cell was observed, and compared with control group. Results Compared to control group, ribozyme can significantly inhibit telomerase activity and cell proliferation of GBC-SD cell. After ribozyme action, the cell ratio of G0/G1 markedly increased, and the cell ratio of S phase markedly decreased. Cellular cleavage and proliferation was markedly inhibited. The apoptosis rate was 17. 10 % and 31 . 01% on day 10 and day 1 3 after ribozyme action. Conclusions Ribozyme can inhibit telomerase activity of GBC-SD cells effectively and induce cell apoptosis.

关 键 词：胆囊癌细胞株 基因疗法 核酶切割技术 端粒酶 抑制作用 

分 类 号：R735.8[医药卫生—肿瘤] Q522[医药卫生—临床医学]