肝细胞生长因子/c-Met信号通路降低体外肝细胞癌细胞对多柔比星的敏感性

Hepatocyte growth factor/c-Met signaling pathway decreases doxorubicin sensitivity in hepatocellular carcinoma in vitro

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.