Hiwi基因靶向沉默对乳腺癌MCF-7/ADM细胞化疗敏感性的影响

目的：探讨耐阿霉素乳腺癌MCF-7/ADM细胞应用Hiwi基因靶向沉默技术后对阿霉素敏感性的变化，阐明Hiwi基因在乳腺癌MCF-7/ADM细胞耐药机制中的作用。方法：MCF-7/ADM细胞分为对照组、Hiwi siRNA1组和Hiwi siRNA2组，分别转染Hiwi control、Hiwi siRNA1和Hiwi siRNA2 3种不同载体，应用实时定量PCR和Western blotting法检测Hiwi基因mRNA和蛋白表达水平，判断转染效率。转染后各组乳腺癌MCF-7/ADM细胞加入不同浓度阿霉素（0、0.1、0.5和1.0mg·L^-1），0mg·L^-1阿霉素组为对照组，采用MTT法检测各组MCF-7/ADM细胞的存活率，即耐药敏感性。结果：与对照组比较，转染后Hiwi siRNA1和Hiwi siRNA2组细胞中Hiwi基因mRNA和蛋白表达水平均明显降低（P<0.01）。与对照组比较，经不同浓度阿霉素作用后，Hiwi siRNA1组和Hiwi siRNA2组细胞存活率明显降低（P<0.01），阿霉素浓度为1.0 mg·L^-1时，2组细胞存活率分别为（48.15±6.28）%和（41.73±6.17）%，对阿霉素的敏感性明显增强。结论：Hiwi基因在MCF-7/ADM细胞中具有高效的靶向沉默，Hiwi基因沉默后的MCF-7/ADM细胞对阿霉素恢复敏感性。

Influence of Hiwigene targeting silence in chemotherapy sensitivity of breast cancer MCF-7/ADM cells

Objective:To explore the changes of chemotherapy sensitivity of breast cancer MCF-7/ADM cells after Hiwi gene targeting silence, and to illuminate the role of Hiwi gene in resistant mechanism of breast cancer MCF-7/ADM cells.Methods:The MCF-7/ADM cells were transfected with Hiwi control,siRNA1 Hiwi gene and siRNA2 Hiwi gene as Hiwi control group, Hiwi siRNA1 group and Hiwi siRNA2 group.The expression levels of Hiwi gene mRNA and protein in breast cancer MCF-7/ADM cells were detected by Real-time PCR and Western blotting methods to judge the transfection rates in various groups.After transfection,the breast cancer MCF-7/ADM cells in various groups were treated with the different concentrations(0,0.1,0.5 and 1.0 mg·L-1) of adriamycin, and the cell resistant sensitivities were detected by MTT method.Results:Compared with control group(0 mg·L-1 adriamycin), the expression levels of Hiwi gene mRNA and protein inMCF-7/ADM cells in Hiwi siRNA1 and Hiwi siRNA2 groups were significantly decreased (P<0.01).Compared with control group, the survival rates of Cells in Hiwi siRNA1 and Hiwi siRNA2 groups were significantly decreased after treated with different concentrations of adriamycin,and the survival rates in Hiwi siRNA1 and Hiwi siRNA2 groups were significantly decreased(P<0.01);they were (48.15±6.28)% and (41.73±6.17)% when the concentration of adriamycin was 1 mg·L-1,and the sensitivities to the adriamycinwere obviously enhanced(P<0.01).Conclusion:The target silencing Hiwi genes in MCF-7/ADM cells is efficient.The sensitivity of MCF-7/ADM cells to adriamycin restores after Hiwi gene silencing.