【摘要】 目的探讨mTOR蛋白与宫颈癌发生、发展过程的相关性和mTOR siRNA对宫颈癌细胞Hela中mTOR、4EBP1表达及细胞侵袭力的影响。方法应用免疫组织化学方法检测宫颈鳞癌组织、宫颈上皮内瘤变(CIN)组织和正常宫颈鳞状上皮组织中mTOR蛋白的表达情况;应用RNAi技术处理宫颈癌Hela细胞,运用免疫细胞化学、原位杂交等技术检测处理前后Hela细胞中mTOR、4EBP1蛋白及mRNA的表达情况;通过Boyden chamber体外侵袭实验,了解mTOR siRNA对Hela细胞侵袭力的影响。结果 mTOR蛋白在30例正常宫颈上皮组织、20例CIN组织和45例宫颈鳞癌组织中的阳性表达率分别为36.7%(11/30)、55.0%(11/20)和71.1%(32/45),正常宫颈组和CIN组与宫颈鳞癌组比较差异有统计学意义(P<0.05);三种不同浓度的mTOR siRNA分别转染宫颈癌Hela细胞24h、48h、72h,各组与各对照组相比,mTOR、4EBP1蛋白及mRNA的表达差异均有统计学意义(P均<0.05);转染mTOR siRNA的Hale细胞,穿越Matrigel的细胞数比各对照组明显减少,差异有统计学意义(P<0.05)。结论 mTOR蛋白的过表达可能与宫颈鳞癌的发生、发展有关;mTOR siRNA可下调宫颈癌Hela细胞中mTOR蛋白及mRNA的表达及抑制宫颈癌细胞的侵袭能力。更多还原

【Abstract】 Objective To explore the relationship between mTOR protein and cervical carcinoma development process, and the effects of mTOR siRNA on mTOR,4EBP1 expression in cervical carcinoma cells and invasion ability of human Hela cells. Methods The expression of mTOR protein were detected by immunohistochemistry in tissues of cervical carcinoma,CIN( cervical intraepithelium neoplasm) and normal cervical squamous epithelium; In situ hybridization and immunohistochemical technique were used respectively to assay the expression of mTOR,4EBP1 mRNA and protein in Hela cells after treatment with RNAi. Boyden chamber experiment in vitro was used to detect the changes of invasion ability of human Hela cells before and after the transfection. Results The positive expression of mTOR protein in 30 cases of normal cervical epithelium,20 cases of CIN and 45 cases of cervical carcinoma were 36. 7%( 11 /30),55%( 11 /20) and 71. 1%( 32 /45) respectively. And there was a significantly higher level of mTOR protein expression in cervical carcinoma when compared with that of in normal cervical squamous epithelium and CIN( P < 0. 05); Then the expression of mTOR, 4EBP1 protein and mRNA in Hela cells were assayed at 24h,48h,72h after treatment with three different concentrations of mTOR siRNA,and results show that the expression of mTOR,4EBP1 mRNA and protein are all significantly downregulated when compared with their respective untreated controls( P < 0. 05); Compared with all the control groups,the number of cells traversed Matrigel is decreased obviously in experiment group,and there is a significant difference( P < 0. 05). Conclusion The mTOR protein were overexpressed in cervical carcinoma,it may be related with the occurrenceand development of cervical carcinoma; mTOR siRNA can inhibit the expression of mTOR mRNA and protein in cervical carcinoma cells,and also their corresponding downstream mole-cule 4EBP1,and mTOR siRNA inhibited the invasion ability of cervical cancer Hela cells.更多还原