在肝癌细胞SK-Hep1中沉默STAT3基因增强sorafenib疗效的初步研究

发布时间：2018-03-21 05:55

本文选题：肝癌　切入点：Sorafenib　出处：《中国生物工程杂志》2017年12期 　论文类型：期刊论文

【摘要】：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SKHep1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-sh STAT3。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Cyclin D1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。
[Abstract]:Objective: to study the role of STAT3 in the resistance of hepatocellular carcinoma cell line SK-Hep1 to sorafenib. To explore the role of STAT3 gene silencing in enhancing the efficacy of sorafenib hepatocarcinoma methods: using shRNA based gene silencing technique to knock down STAT3 in SKHep1 cells and to detect cell growth and sensitivity to sorafenib by CCK8 assay. Western blotting was used to detect the expression of STAT3pSTAT3Y705 and its downstream protein. Results: STAT3 knockout cell line SK-Hep1-sh STAT3 was successfully constructed. The expression of STAT3 protein in the cell was decreased. Cell proliferation was significantly inhibited by the treatment of. Sorafenib down-regulated the phosphorylation level of STAT3 and the expression of downstream proteins Mcl-1 and Cyclin D1. STAT3 gene knockout of SK-Hep1 cells. Conclusion: STAT3 gene silencing based on shRNA can significantly inhibit the proliferation of SK-Hep1 cells and increase the sensitivity of SK-Hep1 cells to sorafenib, which is expected to be a new method to improve the anti-hepatoma effect of sorafenib.
【作者单位】：福建生物工程职业技术学院食品与生物工程系;福建医科大学基础医学院;
【基金】：福建省教育厅中青年教师教育科研项目(JA14410) 福建省科技厅自然科学基金面上项目(2015J01293) 福建省自然科学基金杰青项目(2015J06017) 福建省青年拔尖创新人才福建省科技计划(2014Y2008)资助项目
【分类号】：R735.7

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