UbcH10基因沉默对耐药乳腺癌细胞MCF-7/TXT药物敏感性影响

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

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UbcH10mRNA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UbcH10mRNA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。

Objective To investigate the effects of U bcH10 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 U bcH10. 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 U bcH10 mRNA and protein levels of infected cells were detected by the Real-Time PC R 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 U bcH10 mRNA and protein levels of infected cells significantly decreased.Compared to the control group, the differences were statistically significant（ P〈 0. 05）. Silencing the U bcH10 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 U bcH10 gene can significantly enhance the chemotherapy sensitivity of drug resistant breast cancer cell MCF-7 /TXT towards taxotere.