RNAi干扰HMGB1基因对乳腺癌细胞MCF—7增殖的影响(1)
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[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。
[关键词] 高迁移率族蛋白1；RNA干扰；MCF—7细胞
[中图分类号] R737.9 [文献标识码] A [文章编号] 1673—9701(2012)24—0023—03
Effects of HMGB1 silence by RNA interference on the cell proliferation in MCF—7 cells
LAI Lei1 YANG Linjun2 ZHAI Changlin1
1.Department of Oncology，the First People''s Hospital of Tongxiang City in Zhejiang Province，Tongxiang 314500，China；2.Department of Oncology，Changhai Hospital of Shanghai，Shanghai 200433，China
[Abstract] 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 expression， WB to detect HMGB1 protein expression；the cell proliferating activity in vitro was examined by MTT analysis. Results 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 ......
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