pcdna31-mtor质粒体外转染对乳腺癌细胞生长 - 南方医科大学学报.pdf

·292 · doi 10.3969/j.issn. 1673-4254.2015.02.29 J South Med Univ, 2015, 35(2): 292-294 基础研究 pcDNApcDNA33..11-mTOR-mTOR 质粒体外转染对乳腺癌细胞生长的影响质粒体外转染对乳腺癌细胞生长的影响 刘民锋，郭昭泽，董建宇，杨翼鹏，嵇 健，刘润奇，延 艳，叶长生 南方医科大学南方医院乳腺科，广东 广州 510515 摘要：目的研究信号转导通路mTOR 体外对乳腺癌MCF-7 细胞生长的抑制作用，及对抑癌基因PTEN 的负反馈调节的影响。 方法 用pcDNA3.1-mTOR 真核表达质粒及空载质粒转染MCF-7 ；Western blot 检测转染后mTOR 蛋白的表达；流式细胞技术检 测细胞周期和细胞凋亡情况；检测转染后PTEN 蛋白的表达。结果 转染组细胞的生长速度明显增快，而未转染组和转染空载 体质粒组无明显变化。转染mTOR 基因后，PTEN 蛋白的表达明显下降。结论 mTOR 可能通过PI3K/AKT/PTEN 与mTOR 信 号转导通路负反馈调节PTEN 的表达。mTOR 的增高可促进乳腺癌细胞的生长，为mTOR 的特异性的抑制剂的运用提供了理 论证据。 关键词：mTOR ；信号转导通路；乳腺癌 Effect of mTOR plasmid transfection on growth of breast cancer MCF-7 cells in vitro LIU Minfeng, GUO Zhaozhe, DONG Jianyu, YANG Yipeng, JI Jian, LIU Runqi, YAN Yan, YE Changshen Department of Breast, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China Abstract: Objective To investigate the effect of mTOR signal transduction pathway and down-regulating anti-oncogene PTEN on the growth of breast cancer MCF-7 cells. Methods MCF-7 cells were transfected with the eukaryotic expression plasmid pcDNA3.1-mTOR and non-loaded plasmid, and the expression of mTOR in the cells was detected using Western blotting. Flow cytometry was used to analyze apoptosis and cell cycle of the transfected cells, and the expression of PTEN was detected after transfection. Results The cells transfected with pcDNA3.1-mTOR showed a increased growth rate than those transfected with the non-loaded plasmid and those without transfection. The expression of the protein PTEN decreased obviously in the cells after mTOR trasnfection. Conclusion mTOR can regulate the expression of PTEN via PI3K/AKT/PTEN pathways through a negative feedback mechanism. Increased mTOR expression promotes MCF-7 cell growth, suggesting the potential value of mTOR specific inhibitor in the treatment of breast