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中国生物工程杂志

CHINA BIOTECHNOLOGY
中国生物工程杂志
中国生物工程杂志  2017, Vol. 37 Issue (12): 8-13    DOI: 10.13523/j.cb.20171202
研究报告     
在肝癌细胞SK-Hep1中沉默STAT3基因增强sorafenib疗效的初步研究*
谢琳娜1,2,曾燕华2,柯伙钊1,何文胜1,郑敏2,林德馨2*()
1 福建生物工程职业技术学院食品与生物工程系 福州 350003
2 福建医科大学基础医学院 福州 350108
A Preliminary Study on Enhancing the Efficacy of Sorafenib by Silencing STAT3 Gene in SK-Hep1 Cells
Lin-na XIE1,2,Yan-hua ZENG2,Huo-zhao KE1,Wen-sheng HE1,Min ZHENG2,De-xin LIN2*()
1 Department of Food and Bioengineering, Fujian Vocational College of Bioengineering,Fuzhou 350003, China
2 College of Basic Medical Sciences,Fujian Medical University, Fuzhou　350108, China
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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。
关键词： 肝癌;  Sorafenib;  STAT3;  基因沉默    
Abstract: Objective:

To determine the effect of STAT3 on sorafenib resistance in hepatocarcinoma cell SK-Hep1, and the role of STAT3 gene silencing on enhancing the efficacy of sorafenib in liver cancer.

Methods:

STAT3 gene was knocked down by short hairpin RNA based gene-silencing technology in SK-Hep1 cells. The cell proliferation and the sensitivity to sorafenib were detected by CCK8 assay. The expression levels of STAT3, p-STAT3(Y705),p-STAT3(S727)and its downstream proteins were detected by Western blot.

Results:

STAT3 knockdown cell line SK-Hep1-shSTAT3 was successfully constructed. In these cells, the level of STAT3 expression was decreased; the cell proliferation was significantly suppressed.Sorafenib treatment reduced the level of STAT3 phosphorylation and its downstream protein CyclinD1 and Mcl-1.Sorafenib resistance was reduced in STAT3 knockdown cells.

Conclusion:

Silencing STAT3 by RNA interference can significantly suppress SK-Hep1 cell proliferation, and enhance the sensitivity of liver cancer cells to sorafenib.The data indicates that STAT3 knockdown might be a new way to improve the efficacy of sorafenibin liver cancer.
Key words: Hepatocarcinoma    Sorafenib    STAT3    Gene silence
收稿日期: 2017-08-18 出版日期: 2017-12-16
ZTFLH:  R963  
基金资助: * 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)
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谢琳娜
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林德馨

引用本文:

谢琳娜,曾燕华,柯伙钊,何文胜,郑敏,林德馨. 在肝癌细胞SK-Hep1中沉默STAT3基因增强sorafenib疗效的初步研究*[J]. 中国生物工程杂志, 2017, 37(12): 8-13.

Lin-na XIE,Yan-hua ZENG,Huo-zhao KE,Wen-sheng HE,Min ZHENG,De-xin LIN. A Preliminary Study on Enhancing the Efficacy of Sorafenib by Silencing STAT3 Gene in SK-Hep1 Cells. China Biotechnology, 2017, 37(12): 8-13.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20171202    或    https://manu60.magtech.com.cn/biotech/CN/Y2017/V37/I12/8

图1  Western blot检测STAT3基因沉默的SK-Hep1细胞中STAT3蛋白表达及其磷酸化修饰
图2  不同浓度sorafenib对SK-Hep1细胞STAT3蛋白表达及其磷酸化修饰的影响
图3  Sorafenib对敲减STAT3基因的SK-Hep1细胞活力的影响
图4  SK-Hep1细胞中,sorafenib对STAT3及其下游蛋白Mcl-1、CyclinD1表达的影响
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