摘要
目的 研究miR-199a在卵巢癌OV2008和C13＊细胞对顺铂敏感性的影响,探讨miR-199a调控mTOR在卵巢癌顺铂耐药可能的机制.方法 使用实时定量PCR检测OV2008和C13＊细胞中mTOR的表达.运用免疫印迹检测OV2008和C13＊细胞中mTOR的表达.经转染miR-199a的mimics和inhibitor,实时定量PCR检测OV2008和C13＊细胞中mTOR的变化;免疫印迹检测OV2008和C13＊细胞中mTOR的变化.构建mTOR的荧光素酶载体,在OV2008细胞转染miR-199a的mimics检测其对下游靶基因的调控.结果 实时定量PCR检测结果表明,mTOR在C13＊的表达明显高于OV2008细胞（P＜0.05）.免疫印迹检测结果表明,mTOR在C13＊细胞的表达明显高于OV2008细胞（P＜0.05）.实时定量PCR和免疫印迹检测结果显示,转染miR-199a的mimics抑制mTOR的表达,转染miR-199a的inhibitor促进mTOR的表达.在OV2008细胞转染miR-199a的mimics和mTOR的荧光素酶载体检测结果显示miR-199a对下游靶基因是负调控的.结论 miR-199a通过mTOR调控卵巢癌细胞对顺铂的敏感性,参与卵巢癌细胞的铂类耐药.
Objective To study the effect of miR-199a on the sensitivity of OV2008 and C13 ＊ cells to cisplatinand investigate its mechanism of action.Methods Real-timepolymerase chain reaction （PCR） and Western blot were applied to detect the expressionlevel changes of mammalian target of rapamycin （mTOR） in OV2008 and C13 ＊ cells before and after transfecting the cells with the mimicsand inhibitor of miR-199a with Lipofectamine 2000.We constructed luciferase reporter vectors of mTOR and then transfected OV2008 cell swith the mimics of miR-199a with Lipofectamine 2000 to study the regulation of miR-199a＆#39;s downstream target genes.Results The expression level of mTOR was significantly higher in C13 ＊ than OV2008 cells （P 〈 0.05）.After OV2008 and C13 ＊ cells were transfected with the mimics and inhibitorof miR-199a,real-time PCR and Western blot showed that the mimics of miR-199a repressed the expression of mTOR; and the inhibitor of miR-199a promoted the expression of mTOR.After transfecting OV2008 cells with the mimics of miR-199a and luciferase reporter vectors of mTOR,renillaluciferase reporter gene analysis indicated the expression level of mTOR was negatively regulated by miR-199a.Conclusion miR-199a regulates the sensitivity of ovarian cancer cells to cisplatin through modulating expression of mTOR,and involves in the cisplatin resistance process of ovarian cancer cells.
出处
《中华医学杂志》
CAS
CSCD
北大核心
2015年第33期2705-2708,共4页
National Medical Journal of China
基金
湖北省自然科学基金（2012FFC125）
宜昌市科技研究与开发项目（A12301-08）
关键词
卵巢癌
微小RNA
MTOR
耐药
化疗
Ovarian cancer
microRNA
mTOR
Drug resistance
Chemotherapy
