SiRNA联合介导Livin和Survivin基因沉默诱导人肾癌细胞786-O化疗敏感性的实验研究

陈小刚; 罗鹏程; 张青汉; 袁平成; 姜卫东; 吴尚君; 黄恩应

doi:10.13201/j.issn.1001-1420.2014.06.028

SiRNA联合介导Livin和Survivin基因沉默诱导人肾癌细胞786-O化疗敏感性的实验研究

- 1. 黄石市中心医院泌尿外科(湖北黄石, 435000);
- 2. 肾脏疾病发生与干预湖北省重点实验室

详细信息

通讯作者: 罗鹏程,E-mail:pengcheng.luo@hotmail.com

中图分类号: R737.11

收稿日期: 2014-02-16

Experimental study of chemotherapy sensitivity in human renal carcinoma cell line 786-O induced by siRNA-mediated co-silencing of Livin and Survivin genes

- 1. Department of Urology, Huangshi Center Hospital, Huangshi, Hubei, 435000, China;
- 2. Hubei Key Laboratory of Kidney Disease Pathogenesis and Intervention

More Information

Corresponding author: LUO Pengcheng, E-mail: pengcheng.luo@hotmail.com

Received Date: 16 February 2014

摘要

摘要: 目的:观察siRNA联合介导Livin和Survivin基因的沉默诱导人肾癌细胞786-O化疗敏感性的作用。方法:构建Livin和Suvivin联合靶向的小干扰RNA(siRNA)重组表达载体shRNA,然后将目的载体转染于人肾癌细胞株786-O。应用实时荧光定量聚合酶链反应(Real-time PCR)及Western blot方法分别检测转染前细胞经顺铂、5-FU和丝裂霉素处理后的Livin和Survivin基因的mRNA与蛋白水平的变化。应用CCK-8试验检测转染前后细胞对顺铂、5-氟尿嘧啶(5-FU)和丝裂霉素的半数致死量(IC50)、细胞增殖的变化。结果:CCK-8结果显示,联合介导Livin和Survivin基因沉默组细胞较分别单独介导细胞组化疗的敏感性明显增强(P<0.05),且转染前后细胞对顺铂、5-FU和丝裂霉素的IC50发生显著变化(P<0.05)。结论:SiRNA联合介导Livin和Survivin基因表达下调,发挥协同增强的siRNA作用。
- 肾癌 /
- siRNA /
- Livin基因 /
- Survivin基因 /
- 786-O细胞
Abstract: Objective:To investigate the chemotherapy sensitivity of human renal carcinoma cell line 786-O induced by siRNA-mediated co-silencing of Livin and Survivin genes.Method:We constructed Livin and Survivin combined targeting small interfering RNA (siRNA) recombinant expression vector. Then we transfected them in 786-O cell line. Real-time fluorescent quantitative polymerase chain reaction (Real-time PCR) and Western blot methods were used respectively to detect the expression level of Livin and Survivin in mitomycin, 5-FU and cisplatin treated 786-O cell. Then we investigated the effect of these shRNA in human renal carcinoma cell line 786-O, including the affection of the chemotherapy sensitivity and proliferation.Result:CCK-8 analysis results showed that the sensitivity to chemotherapy of combined mediated Livin and Survivin genes silencing were higher than separately mediated Livin and Survivin (P<0.05). Moreover, IC50 showed significant difference between before and after the transfection (P<0.01). Conclusion:SiRNA-mediated down-regulation of Livin and Survivin genes enhance the synergistic effect.
- renal cancer /
- siRNA /
- Livin gene /
- Survivin gene /
- 786-O cell

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