上海交通大学学报（医学版）

上海交通大学学报（医学版）

• 论著（基础研究） • 上一篇    下一篇

UbcH10基因沉默对耐药乳腺癌细胞MCF-7/TXT药物敏感性影响

汪 成1，单 鸣1*，潘贇昊1，徐 明2，包家林1   

  1. 上海市黄浦区中心医院 1.乳腺外科， 2.病理科， 上海 200002
  • 出版日期:2014-10-28 发布日期:2014-10-28
  • 作者简介:汪成（1975—）， 男， 副主任医师， 硕士; 电子信箱: wangcheng1212@hotmail.com; 单鸣（1976—）， 男， 主治医师， 学士; 电子信箱: shanming99@gmail.com。*为共同第一作者
  • 基金资助:

    上海市医学重点专科建设项目（ZK2012A13）

Effects of UbcH10 gene silencing on drug sensitivity of drug resistant breast cancer cell MCF-7/TXT

WANG Cheng1, SHAN Ming1*, PAN Yun-hao1, XU Ming2, BAO Jia-lin1   

  1. 1.Department of Breast Surgery, 2.Department of Pathology, Central Hospital of Huangpu District, Shanghai 200002, China
  • Online:2014-10-28 Published:2014-10-28
  • Supported by:

    Construction Project of Shanghai Medical Key Discipline, ZK2012A13

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摘要：

目的 观察UbcH10基因沉默对MCF-7/TXT细胞耐药的影响。方法 设计针对UbcH10的siRNA序列，构建shRNA表达载体并进行重组慢病毒包装，重组病毒感染MCF-7/TXT细胞，转染后72 h通过荧光标记物确认细胞感染效率，收集转染后细胞，Real-Time PCR和Western blotting分别检测转染后细胞内UbcH10 mRNA及蛋白含量变化；CCK-8检测基因沉默对MCF-7/TXT化疗药物敏感性的影响。结果 在MCF-7/TXT细胞中成功进行了UbcH10基因沉默实验，病毒感染后72 h细胞的感染效率达到约90%，细胞内UbcH10 mRNA和蛋白相对含量明显降低，与对照组比较差异具有统计学意义（P<0.05）；UbcH10基因沉默可以明显增强MCF-7/TXT对多西他赛的药物敏感性，多西他赛对基因干预组和对照组细胞的不同时间点IC50值明显降低，差异具有统计学意义（P<0.05）。结论 UbcH10沉默可显著增强乳腺癌耐药细胞MCF-7/TXT对多西他赛的化疗敏感性。

关键词: MCF-7/TXT, siRNA, 慢病毒, UbcH10, 耐药性

Abstract:

Objective To investigate the effects of UbcH10 gene silencing on the drug resistance of human breast cancer cell MCF-7/TXT. Methods The shRNA expression vectors were constructed using the siRNA sequences which were designed based on the coding sequence of UbcH10. MCF-7/TXT cells were then infected by lentivirus. After being infected for 72 h, the infection efficiency was observed through the fluorescent marker and infected cells were collected. The variations of UbcH10 mRNA and protein levels of infected cells were detected by the Real-Time PCR and Western blotting, respectively. The effects of gene silencing on the sensitivity of chemotherapeutic drugs for MCF-7/TXT cells were detected by the CCK-8. Results The gene silencing test of MCF-7/TXT cells was successfully. After being infected for 72 h, the infection efficiency was about 90% and UbcH10 mRNA and protein levels of infected cells significantly decreased. Compared to the control group, the differences were statistically significant (P<0.05). Silencing the UbcH10 gene significantly enhanced the drug sensitivity of taxotere towards MCF-7/TXT cells. IC50 values of taxotere of the gene silencing group at different time points were significantly lower than those of the control group. The differences were statistically significant (P<0.05). Conclusion Silencing the UbcH10 gene can significantly enhance the chemotherapy sensitivity of drug resistant breast cancer cell MCF-7/TXT towards taxotere.

Key words: MCF-7/TXT, siRNA, Lentivirus, UbcH10, drug resistance