靶向mTOR siRNA的合成及对MCF-7细胞mTOR基因表达的影响  

Establishment of siRNA targeted at mTOR and its effect on expression of mTOR gene in human breast carcinoma MCF-7 cells

作　　者：张岳灿[1] 翟娜娜[2] 

机构地区：[1]宁波天一职业技术学院人体形态学教研室,宁波315104 [2]黄河科技学院人体解剖学与组织胚胎学教研室,郑州450052

出　　处：《郑州大学学报（医学版）》2012年第3期361-364,共4页Journal of Zhengzhou University(Medical Sciences)

摘　　要：目的:合成靶向哺乳动物雷帕霉素靶蛋白(mTOR)siRNA,观察其对人乳癌MCF-7细胞mTOR基因表达的影响。方法:以mTOR基因434～452序列为靶点,设计并体外合成mTORsiRNA,用100和150nmol/L的siRNA转染MCF-7细胞24、48、72h,同时设无关对照组(转染非特异性siRNA)和正常对照组(不转染),运用免疫组化SP法和原位杂交技术检测各组细胞mTOR蛋白及mRNA的表达水平。结果:转染24、48和72h后,与各对照组相比,siRNA转染组细胞mTOR蛋白及mRNA的表达均降低(P均<0.05),2个siRNA转染组之间差异无统计学意义(P均>0.05)。不同时间点mTOR蛋白及mRNA表达水平差异无统计学意义(P>0.05)。结论:设计合成的siRNA可有效抑制乳癌MCF-7细胞mTOR基因的表达。Aim：To establish siRNA targeted at mammalian target of rapamycin（mTOR） and observe its effect on the expression of mTOR gene in human breast carcinoma MCF-7 cells.Methods：mTOR siRNA targeted at the 434~452 sequence of mTOR gene was synthesized by transcription in vitro.mTOR siRNA at 100,150 nmol/L was transfected into MCF-7 cells for different time（24,48,and 72 h）,then the expressions of mTOR protein and mRNA in cells were detected by immunocytochemistry and in situ hybridization methods.The cells without transfection or transfected with nonsense siRNA were the control.Results：There were significant differences in the expression of mTOR protein and mRNA among the four groups（P0.05）.There was no statistical differences in mTOR protein and mRNA expression levels between the two siRNA transfected groups（P0.05）,but both of them were higher than those of the control（P0.05）.The expression levels of mTOR protein and mRNA at different time points had no differences（P0.05）.Conclusion：The siRNA targeted at mTOR has been synthesized successfully,and could inhibit the expression of mTOR.

关 键 词：RNA干扰 雷帕霉素靶蛋白 乳癌 

分 类 号：R739.7[医药卫生—肿瘤]