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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

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