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目的:研究表柔比星(epirubicin,EPI)联合川芎嗪(tetramethylpyrazine,TMP)对人肝癌细胞TGF-β1/Smads通路的影响,以探讨联合用药的效果。方法:人肝癌细胞分为4个组,即对照组、EPI6.0μg/m L组、TMP0.2mg/m L组、EPI6.0μg/m L+TMP0.2mg/m L组,各组给予对应药物干预24h,CCK-8细胞计数法测定细胞增殖水平的变化,real-time PCR方法检测各组肝癌细胞TGF-β1、Smad4及Smad7 mRNA表达,western-blot方法检测各组细胞TGF-β1及Smad4蛋白表达。结果:在EPI、TMP及EPI+TMP干预24h后,肝癌细胞的增殖明显受到抑制,肝癌细胞TGF-β1及Smad7的mRNA及蛋白表达明显下调,而Smad4的mRNA及蛋白表达明显上调,联合用药的效果较单一药物效果强(均P<0.05)。结论:EPI及TMP的抗肝癌作用机理之一可能与它们对肝癌细胞TGF-β1/Smads通路的调控有关,联合用药的效果较单一药物效果强。
Objective: To investigate the effect of epirubicin (EPI) combined with tetramethylpyrazine (TMP) on TGF-β1 / Smads pathway in human hepatocellular carcinoma cells in order to explore the effect of combination therapy. Methods: Human hepatocellular carcinoma cells were divided into 4 groups: control group, EPI6.0μg / m L group, TMP0.2mg / m L group, EPI6.0μg / m L + TMP0.2mg / m L group, The changes of cell proliferation were detected by CCK-8 cell counting after 24 h of drug intervention. The mRNA expressions of TGF-β1, Smad4 and Smad7 in each group were detected by real-time PCR. The expressions of TGF-β1 and Smad4 Protein. RESULTS: After treated with EPI, TMP and EPI + TMP for 24 h, the proliferation of hepatoma cells was significantly inhibited. The mRNA and protein expressions of TGF-β1 and Smad7 in hepatocellular carcinoma cells were significantly down-regulated while the mRNA and protein expressions of Smad4 were significantly up-regulated The effect is better than single drug (all P <0.05). CONCLUSION: One of the mechanisms of anti-hepatocarcinogenesis of EPI and TMP may be related to the regulation of TGF-β1 / Smads pathway in hepatocellular carcinoma cells. The effect of combination therapy is better than that of single drug.