高级搜索

雷帕霉素及mTOR siRNA对宫颈癌HeLa细胞mTOR和4EBP1表达的影响

Effects of mTOR siRNA and Rapamycin on mTOR and 4EBP1 Expressions in Human HeLa Cells

  • 摘要: 探讨mTOR蛋白与宫颈癌发生、发展过程的相关性及雷帕霉素和mTOR siRNA对宫颈癌细胞中mTOR、4EBP1表达的影响。方法 应用免疫组织化学方法检测正常宫颈鳞状上皮组织、CIN组织和宫颈癌组织中mTOR蛋白的表达情况;应用RNAi技术及雷帕霉素分别处理宫颈癌HeLa细胞,运用原位杂交、免疫细胞化学技术分别检测处理前后HeLa细胞中mTOR、4EBP1 mRNA及蛋白的表达情况。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),正常宫颈组织、CIN组织与宫颈癌组织中mTOR阳性表达率比较差异有统计学意义(P<0.05);三种不同浓度的雷帕霉素及mTOR siRNA 分别处理宫颈癌HeLa细胞24、48和72 h后,各组与相应对照组相比,mTOR、4EBP1 mRNA及蛋白的表达量均有显著下降(P均<0.05)。结论 mTOR蛋白在宫颈癌组织中过表达,其过表达可能与宫颈癌的发生、发展过程相关;雷帕霉素和mTOR siRNA可抑制宫颈癌细胞mTOR mRNA及蛋白的表达,同时影响其下游4EBP1分子的表达。

     

    Abstract: Objective To explore the relationship between mTOR protein and cervical carcinoma development process,and the effects of rapamycin and mTOR siRNA on mTOR and 4EBP1 expressions in cervical carcinoma cells. Methods The expressions of mTOR protein were detected by immunohistochemistry in tissues of normal cervical squamous epithelium, cervical intraepithelium neoplasm(CIN) and cervical carcinoma. In situ hybridization and immunohistochemistry were used respectively to analyze the expressions of mTOR, 4EBP1 mRNA and protein in HeLa cells before and after treatment with rapamycin or RNAi. Results The positive expressions 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 were a signifi cantly higher levels of mTOR protein expression in cervical carcinoma than those in normal cervical squamous epithelium and CIN ( P <0.05 ). Compared with the control group, the expressions of mTOR, 4EBP1 mRNA and protein in HeLa cells were signifi cantly downregulated after treatment at three different concentrations of rapamycin or mTOR siRNA for 24, 48 and 72 h( P <0.05 ). Conclusion The overexpression of mTOR protein may be related with the occurrence and development of cervical carcinoma. Rapamycin and mTOR siRNA could inhibit the expressions of mTOR mRNA, protein and corresponding downstream molecule 4EBP1in cervical carcinoma cells.

     

/

返回文章
返回