Hiwi基因靶向沉默对乳腺癌MCF-7/ADM细胞化疗敏感性的影响
Journal of Jilin University(Medicine Edition)(2017)
Abstract
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1.0 mg·L-1),0 mg·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.
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Key words
breast neoplasms,Hiwigene,targeted silence,MCF-7/ADM cells,sensitivity
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