慢病毒介导MIM—B基因小干扰RNA增强索拉非尼对差异表达pERK人肝癌细胞的体外增殖抑制作用  

In vitro inhibitory effect of sorafenib on proliferation of human hepatocellular carcinoma cells with differential pERK expression reinforced by lentivirus-mediated small RNA interference targeting MIM-B gene

作　　者：黄修燕[1] 黄自丽[2] 许永华[2] 艾开兴[1] 汤钊猷[3] 郑起[1] 

机构地区：[1]上海交通大学附属第六人民医院普外科,200233 [2]上海市徐汇区中心医院影像科 [3]复旦大学肝癌研究所

出　　处：《中华实验外科杂志》2011年第3期396-398,共3页Chinese Journal of Experimental Surgery

基　　金：基金项目：国家重点基础研究（973）计划资助项目（2004CB518708）

摘　　要：目的 观察转移消失蛋白B（MIM-B）小干扰RNA（LV-siMTSS1）联用索拉非尼对差异表达磷酸化胞外信号调节激酶（pERK）肝癌（HCC）增殖的影响.方法 构建LV-siMTSS1,检测MIM-B、pERK蛋白表达.用LV-siMTSS1（1.0×107～36.0×107U/nl）、索拉非尼（0.01～30.0μmoL/L）干预细胞,计算IC50.结果 高转移潜能HCC高表达MIM-B.高转移潜能MHCC97L、MHCC97H、HCCLM3、HCCLM6相对表达pERK（0.371、0.636、0.949、1.112）与低转移潜能Hep3B、HLE、SMMC7721（0.115、0.130、0.188）比较差异有统计学意义（P＜0.05）.前者对索拉非尼敏感性（IC50为13.5、12.4、10.2、8.8 μmoL/L）强于后者（IC50为26.4、20.5、18.9 μmol/L,P＜0.05）;LV-siMTSS1增敏索拉非尼药效在低表达pERK的Hep3B中最显著.结论 LV-siMTSS1显著增强低表达pERK的HCC对索拉非尼药物敏感性.Objective To observe in vitro effect of sorafenib combined with lentivirus-mediated small RNA interference targeting the gene of missing in metastasis B （ MIM-B,LV-siMTSS1 ） on proliferation of human hepatocellular carcinoma （HCC） cells with differential phospgorylated extracellular signalregulated kinase （pERK） expression.Methods The expression of MIM-B and pERK was detected in HCC cell lines with differential metastatic potentials.The siRNA targeting MIM-B gene was cloned to one lentivirus work vector.Work vector and three package plasmids were co-transfected into 293T cells with the help of lipefeetamine 2000 （ named LV-siMTSS1 ）.The cultured cell lines were intervened by LV-siMTSS1 （ 1.0 × 107-36.0 × 107） U/ml and （or） sorafenib （0.01-30.0） μmol/L in vitro.Cellular activity was determined by Cell Counting Kit-8 to calculate IC50.Results The basal pERK and MIM-B levels were increased stepwise in cell lines in accordance with their metastatic potential.There was statistically significant difference in the pERK expression was made between the cell lines of MHCC97L,MHCC97H,HCCLM3 and HCCLM6 （0.371,0.636,0.949 and 1.112） with higher metastatic potential and those of Hep3B,HLE and SMMC7721 （0.115,0.130 and 0.188） with lower one （P〈0.05）.Cells with higher pERK expression were more sensitivity （ IC50,13.5,12.4,10.2 and 8.8 μmol/L） than those with lower pERK expression （ IC50,26.4,20.5 and 18.9 μmol/L） to Sorafenib （ P〈0.05 ）.Cells transfected with LV-siMTSS1 were sensitivity to Sorafenib,which was most significant in Hep3B cell line with lower pERK expression.Conclusion Transfection with LV-siMTSS1 reinforced the inhibitory effect of Sorafenib on HCC cell proliferation,especially for cells with lower pERK expression.

关 键 词：癌 肝细胞 索拉非尼 胞外信号调节激酶 转移 

分 类 号：R735[医药卫生—肿瘤]