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目的 观察AFP增强型四元复合体介导的N ras反义RNA转移系统对HBV转基因肝癌细胞系HepG2 .2 .15致瘤性的体内外抑制作用。方法 构建含有N ras反义RNA的AFP增强型四元复合体 ,体外瞬时转染HBV转基因HepG2 .2 .15细胞 ,流式细胞术检测转染前后N ras蛋白表达水平、细胞凋亡率及细胞周期的变化 ,同时建立稳定表达N ras反义RNA的肝癌细胞系HepG2 .2 .15 /as ras,绘制转染前后生长曲线。体内抑瘤试验分别以HepG2 .2 .15或HepG2 .2 .15 /as ras细胞制备荷瘤裸鼠模型 ,比较二者成瘤率。HepG2 .2 .15成瘤组局部瘤内注射N ras反义RNA四元复合体 ,研究其对肿瘤的抑制作用。结果 AFP增强型四元复合体介导N ras反义RNA体外瞬时转染HepG2 .2 .15细胞可显著降低胞内N ras蛋白的表达水平 (P <0 .0 5 ) ,使细胞生长停滞于G0 /G1期 ,且可诱导细胞凋亡。体内抑瘤实验显示 ,HepG2 .2 .15 /as ras细胞注射组成瘤率 ( 4 0 % )显著低于HepG2 .2 .15细胞注射组( 10 0 % )。瘤内注射N ras反义RNA四元复合体可使肿瘤体积明显缩小。结论 AFP增强型四元复合体介导的N ras反义RNA转移系统可降低HBV转基因肝癌细胞HepG2 .2 .15N ras蛋白的表达水平 ,逆转其恶性行为 ,体内外均可有效抑制肿瘤的生长增殖
Objective To investigate the in vivo and in vitro inhibition of HepG2 .2.15 tumorigenicity induced by HBV AFP-enhanced quaternary complex-mediated N ras antisense RNA transfer system. Methods AFP-enhanced quaternary complex containing N ras antisense RNA was constructed and transiently transfected into HBV transgenic HepG2.2.15 cells in vitro. Flow cytometry was used to detect the expression of N ras protein, the apoptosis rate and the percentage of cells Cycle, and establish a HepG2.2.15 / as ras stably expressing Nsr antisense RNA. The growth curve before and after transfection was drawn. In vivo antitumor experiment HepG2.2.15 or HepG2.2.15 / as ras cells were used to prepare tumor-bearing nude mice models, respectively. HepG2 .2 .15 into the tumor group local intratumoral injection of Ns antisense RNA quaternary complex to study its inhibitory effect on the tumor. Results Transfection of AFP-enhanced quaternary complex with NsRNA antisense RNA in HepG2.2.15 cells could significantly decrease the expression of intracellular N ras protein (P <0.05), and cell growth was arrested at G0 / G1 phase, and can induce apoptosis. In vivo antitumor experiments showed that the injection rate of HepG2.2.15 / as ras cells (40%) was significantly lower than that of HepG2.2.15 cells (10%). Intratumor injection of NSRA antisense RNA quaternary complex can significantly reduce the tumor volume. Conclusion AFP-enhanced quaternary complex-mediated N ras antisense RNA transfer system can reduce the expression level of HepG2.215N ras in HBV transgenic hepatoma cells and reverse its malignant behavior, both in vitro and in vivo, and can effectively inhibit tumor growth and proliferation