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目的:研究STAT3在肝癌细胞SK-Hep1对sorafenib抗性中的作用,并探讨STAT3基因沉默在增强sorafenib肝癌疗效中的作用。方法:应用基于shRNA的基因沉默技术在肝癌细胞SK-Hep1中敲减STAT3;用CCK8法检测细胞的生长情况与对sorafenib的敏感性;蛋白印迹法(Western blot)检测STAT3、p-STAT3(Y705)、p-STAT3(S727)以及其下游蛋白的表达变化。结果:成功构建了STAT3敲减的细胞株SK-Hep1-shSTAT3。该细胞中STAT3蛋白表达降低,细胞增殖明显受到抑制。Sorafenib的处理下调了STAT3的磷酸化水平及其下游蛋白Mcl-1和CyclinD1的表达。STAT3基因敲减的SK-Hep1细胞,对sorafenib的敏感性增强。结论:基于shRNA的STAT3基因沉默能明显抑制SK-Hep1细胞增殖,提高细胞对sorafenib的敏感性,有望成为提高sorafenib抗肝癌疗效的一种新手段。
AIM: To investigate the role of STAT3 in sorafenib resistance in SK-Hep1 hepatocellular carcinoma cells and to explore the role of STAT3 gene silencing in enhancing sorafenib liver cancer. METHODS: STAT3 was knocked down in SK-Hep1 cells by shRNA-based gene silencing technique. Cell growth and sensitivities to sorafenib were determined by CCK8 assay. STAT3 and p-STAT3 were detected by Western blot ), P-STAT3 (S727) and its downstream proteins. Results: STAT3 knockdown cell line SK-Hep1-shSTAT3 was successfully constructed. STAT3 protein expression decreased in this cell, cell proliferation was significantly inhibited. Sorafenib treatment down-regulated the phosphorylation of STAT3 and the expression of its downstream proteins, Mcl-1 and CyclinD1. STAT3 knockdown of SK-Hep1 cells, sorafenib sensitivity increased. Conclusion: The shRNA-based STAT3 gene silencing can significantly inhibit the proliferation of SK-Hep1 cells and increase the sensitivity of sorafenib to cells, which is expected to be a new method to improve the therapeutic effect of sorafenib on hepatocellular carcinoma.