摘要
目的探讨高迁移率族蛋白1(high mobility group box 1,HMGB1)siRNA干扰后对乳腺癌细胞MCF-7增殖的影响。方法构建靶向HMGB1 mRNA的质粒载体pRI-GFP-1、pRI-GFP-2以及阴性对照载体pRI-GFP-Neg,分别转染乳腺癌细胞MCF-7,48 h、72 h后免疫印迹法(Western blot)检测HMGB1基因蛋白表达,噻唑蓝(MTT)比色法检测体外增殖活性。结果转染后,与空质粒组pRI-GFP-Neg相比,pRI-GFP-1组、pRI-GFP-2组MCF-7细胞HMGB1蛋白表达下降,MTT显示干扰组细胞增殖速率与质粒对照及空白对照组相比显著降低。结论应用RNAi技术可以显著干扰HMGB1蛋白的表达,进而有效抑制MCF-7细胞的增殖活性。
Objective To investigate the effect of HMGB1 RNA interference on MCF-7 cell proliferation. Methods The plasmid construct pRI-GFP-1, pRI-GFP-2 targeted HMGB1 mRNA and negative control construct pRI-GFP-Neg, were transfected into MCF-7 cells. After 48 h, 72 h, using real-time quantitative PCR to detect HMGB1 gene mRNA expres sion, WB to detect HMGB1 protein expression;the cell proliferating activity in vitro was examined by MTT analysis. Re suits After transfection, compared with the pRI-GFP-Neg group, the inhibition rates of HMGB1 expression in MCF-7 cells in pRI-GFP-1 group, pRI-GFP-2 group were decreased. The cell proliferation rate was significantly lower in pRI-GFP-1 group than in pRI-GFP-Neg group （P 〈 0.05）. Conclusion Application of RNAi technology can significantly interfere the HMGB1 gene expression, thus inhibit MCF-7 cell proliferation.
出处
《中国现代医生》
2012年第24期23-25,共3页
China Modern Doctor
