摘要
目的 用合成的c-Met反义寡核苷酸链,构建的c—Met反义质粒及U1SnRNA/核酶/反义c-Met复合体来封闭c-Met信号传导,探讨c-Met信号传导阻滞对肝癌生长和转移的影响。方法 以脂质体法将c-Met反义质粒、U 1SnRNA/核酶/反义c—Met复合体转染肝癌SF7721细胞,分别用四甲基偶氮唑蓝、生长曲线、穿膜试验观察转染前后细胞的变化。将转染前后细胞分别接种裸鼠,观察移植瘤生长及转移情况。结果c-Met反义寡核苷酸链能抑制肝癌SF7721细胞的增殖,t=3.58,P< 0.05,c-Met反义核酸质粒及U1SnRNA/核酶/反义c-Met复合体质粒转染后细胞表达受体c—Met的量均减少,且转染后细胞增殖明显减慢,为对照组的50％,F=4.87,P<0.05,侵袭运动能力减弱。初步动物实验显示转染后细胞生长较慢,潜伏期延长,35 d后反义(6.37 g)与核酶(6.14 g)实验组肿瘤生长明显低于对照组(11.01g),F=5.17,P<0.05。结论 c-Met受体基因表达在肝癌生长转移中起重要作用,阻断c-Met—SF的信号传导降低肿瘤生长甚至转移的能力。
Objective To explore the influence of c-Met inhibitor by synthetic c-Met antisense oligonucleotide, constructive c-Met antisense plasmid and the complex plasmid of UlSnRNA/ ribozyme/anti-Met on the growth and metastasis of hepatocellular carcinoma cells. Methods Gene transfection was operated by Lipofectin on SF7721 cells. The difference of the cells before and after transfection was compared by MTT, growth curves and transwell test in vitro. In vivo, the cells before and after transfection were implanted subcutaneously into nude mice respectively to observe tumor growth and metastasis. Results C-Met antisense oligonucleotide could inhibit the growth of hepatocellular carcinoma SF7721 cells (t = 3.58, P < 0.05). After transfection, the expression of c-Met protein decreased. Growth curves showed that the cells after transfection proliferated more slowly, about 50% of control cells (F = 4.87, P < 0.05), and their motility and invasiveness decreased, compared with those before transfected. In vivo experiment, tumors originated from c-Met antisense oligonucleotide treated cells and the antisense/ribozyme/Ul SnRNA treated cells grew more slowly (about 54.5% of those from the control cells), and the latent prolonged. After 35 days, the average weight of tumors in the two group nude mice were lighter than that in the control group nude mice (F = 5.17, P < 0.05). Conclusion Inhibition of c-Met expression by c-Met antisense oligonucleotide and the complex of antisense/ribozyme/UlSnRNA can inhibit the growth and metastasis of SF7721 hepatocarcinoma cells in vitro and in vivo.
出处
《中华肝脏病杂志》
CAS
CSCD
2003年第8期487-489,共3页
Chinese Journal of Hepatology
基金
国家自然科学基金(39970290)
