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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 high mobility group box 1 (HMGB1) siRNA on the proliferation of MCF-7 breast cancer cells. Methods The plasmid vector pRI-GFP-1, pRI-GFP-2 targeting HMGB1 mRNA and negative control vector pRI-GFP-Neg were constructed and transfected into breast cancer cell MCF-7 for 48 h. Western Blot Western blot was used to detect the protein expression of HMGB1. MTT assay was used to detect the proliferation activity in vitro. Results Compared with pRI-GFP-Neg, the expression of HMGB1 in pRI-GFP-1 group and pRI-GFP-2 group decreased after transfection. MTT showed that the proliferation rate of MCF- Compared with the blank control group, it decreased significantly. Conclusion RNAi can significantly interfere with the expression of HMGB1 protein, and then effectively inhibit the proliferation activity of MCF-7 cells.