CAJ | 학술논문

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.
목적 관찰전이소실단백B(MIM-B)소간우RNA(LV-siMTSS1)련용색랍비니대차이표체린산화포외신호조절격매(pERK)간암(HCC)증식적영향.방법 구건LV-siMTSS1,검측MIM-B、pERK단백표체.용LV-siMTSS1(1.0×107～36.0×107U/nl)、색랍비니(0.01～30.0μmoL/L)간예세포,계산IC50.결과 고전이잠능HCC고표체MIM-B.고전이잠능MHCC97L、MHCC97H、HCCLM3、HCCLM6상대표체pERK(0.371、0.636、0.949、1.112)여저전이잠능Hep3B、HLE、SMMC7721(0.115、0.130、0.188)비교차이유통계학의의(P＜0.05).전자대색랍비니민감성(IC50위13.5、12.4、10.2、8.8 μmoL/L)강우후자(IC50위26.4、20.5、18.9 μmol/L,P＜0.05);LV-siMTSS1증민색랍비니약효재저표체pERK적Hep3B중최현저.결론 LV-siMTSS1현저증강저표체pERK적HCC대색랍비니약물민감성.
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.