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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。
Objective To investigate the relationship between the expression of mTOR and carcinogenesis and progression of cervical carcinoma and the effect of mTOR siRNA on the expression of mTOR and 4EBP1 in cervical cancer cells Hela. Methods The expression of mTOR protein was detected by immunohistochemistry in cervical squamous cell carcinoma, cervical intraepithelial neoplasia (CIN) and normal cervical squamous epithelium. The cervical cancer Hela cells were treated with RNAi technique. Immunocytochemistry, Bit hybridization and other techniques before and after Hela cells detection of mTOR, 4EBP1 protein and mRNA expression; Boyden chamber in vitro invasion assay to understand mTOR siRNA on Hela cell invasiveness. Results The positive rates of mTOR protein in 30 cases of normal cervical epithelium, 20 cases of CIN and 45 cases of cervical squamous cell carcinoma were 36.7% (11/30), 55.0% (11/20) and 71.1% (32 / 45). There was significant difference between normal cervix group and CIN group and cervical squamous cell carcinoma group (P <0.05). Three different concentrations of mTOR siRNA were transfected into Hela cells of cervical cancer for 24h, 48h, 72h, Compared with the control group, the expressions of mTOR and 4EBP1 protein and mRNA were all significantly different (all P <0.05). The number of cells transfected with mTOR siRNA in Hale cells and Matrigel decreased significantly compared with the control group Significance (P <0.05). Conclusion The overexpression of mTOR may be related to the occurrence and development of cervical squamous cell carcinoma. MTOR siRNA may down-regulate the expression of mTOR protein and mRNA in Hela cells and inhibit the invasion of cervical cancer cells.