目的 探讨肝细胞生长因子(HGF)/c-Met信号通路体外对多柔比星(DOX)作用肝细胞癌(HCC)细胞株的影响.方法 以不同生物学特性和遗传特征的HCC细胞株Huh7、HepG2、MHCC97-L和MHCC97-H作为研究对象.通过实时定量聚合酶链反应(RT-PCR)检测不同HCC细胞株中c-Met mRNA的表达,通过蛋白印迹法分析不同药物处理下这些细胞株中c-Met和磷酸化Met(p-Met)的表达水平.通过CCK-8实验分析不同细胞株对DOX的敏感性.组间统计学比较采用重复测量的方差分析和t检验.结果 相较于Huh7和HepG2细胞株,MHCC97-L和MHCC97-H细胞株高表达c-Met和p-Met mRNA,并且对DOX不敏感.HGF能够激活Huh7和HepG2细胞株的c-Met信号通路,并降低Huh7和HepG2细胞对DOX的敏感性[对Huh7细胞抑制率:(34.848±5.370)%比(66.409±5.792)%;对HepG2细胞抑制率:(34.351±3.305)%比(62.308±5.453)%;均P=0.002].而使用c-Met抑制剂则增强DOX对MHCC97-L和MHCC97-H细胞的抑制作用[对MHCC97-L细胞抑制率:(73.106±3.472)%比(13.636±4.097)%;对MHCC97-H细胞抑制率:(64.444±4.006)%比(6.296±2.796)%;均P〈0.001].结论 HGF/c-Met信号通路与体外DOX作用HCC细胞的效果密切相关.
Objective To explore the effect of hepatocyte growth factor (HGF)/c-Met signaling in doxorubicin (DOX) treatment of hepatocellular carcinoma (HCC). Methods Different biologic and genetic characteristics human HCC cell lines, Huh7, HepG2, MHCC97-L and MHCC97H were used in this experiment. Variation in c-Met mRNA expression level among different HCC cell lines was analyzed by RT-PCR. Western blot analysis was performed to detect c-Met and p-Met expression levels in these cell lines. CCK-8 experiment was carried to analyze the DOX sensitivity in various cell lines. t test and repeated measure analysis of variance were used for statistical analysis. Results Both c-Met and p-Met were overexpressed in MHCC97-L and MHCC97-H cell lines and these cell lines were resistant to DOX compared to Huh7 and HepG2. However, treatment of HGF in Huh7 and HepG2 cells activated c-Met signaling pathway and decreased the sensitivity of these two cell lines to DOX [inhibition rate: Huh7 (34.848 ±5.370) vs. (66.409±5.792)%, HepG2 (34.351±3.305) %vs. (62.308±5.453) %, both P=0.002]. Whereas administration of c-Met inhibitor in MHCC97-L and MHCC97-H cell lines significantly increased the sensitivity to DOX [inhibition rate: MHCC97-L (73.106 ±3.472) % vs. (13.636 ±4.097) %; MHCC97-H (64.444 ±4.006) % vs. (6.296 ±2.796) %, both P〈 0.001]. Conclusion HGF/c-Met signaling pathway is related the treatment efficacy of DOX in HCC.