MicroRNA-200a/TFAM通路调控紫杉醇对乳腺癌细胞的化疗敏感性

The Function of Paclitaxel on the Breast Cancer Cells Chemosensitivity by Regulating MicroRNA-200a/TFAM Pathway

目的探讨MicroRNA-200a/TFAM在MCF-7细胞对紫杉醇化疗敏感性中的作用。方法分别采用实时荧光定量PCR、蛋白印迹法检测不同浓度紫杉醇处理MCF-7细胞后mi R-200a、TFAM蛋白的表达差异;转染mi R-200a(或antimi R-200a)联合紫杉醇处理MCF-7细胞后,应用四甲基偶氮唑蓝(MTT)法和5-溴脱氧尿嘧啶核苷(Brd U)观察细胞增殖活力的变化;转染pc DNA3.1-TFAM质粒联合紫杉醇处理MCF-7细胞后,MTT和Brd U法检测细胞增殖活力的变化。结果在MCF-7细胞中,mi R-200a的表达量随紫杉醇浓度增加逐渐升高,而TFAM的表达量随紫杉醇浓度升高逐渐下降。紫杉醇与mi R-200a共同作用于MCF-7细胞,可增强紫杉醇对细胞增殖的抑制作用;紫杉醇与anti-mi R-200a共同作用于MCF-7细胞时,可减弱紫杉醇对细胞增殖的抑制作用;而紫杉醇与TFAM共同孵育MCF-7细胞时,可减弱紫杉醇对细胞增殖的抑制作用。结论 mi R-200a及TFAM参与调控紫杉醇对MCF-7细胞的化疗敏感性。

Objective To investigate the function of paclitaxel on chemo-sensitivity of MCF-7cells by regulating miR-200a/TFAMpathway.Methods The mRNA and protein expression of miR-200a and TFAM in MCF-7cells were detected by real-time PCR and Westernblot,respectively.After transfection of MCF-7cells with miR-200a mimics,anti-miR-200a or pcDNA3.1-TFAM plasmids under paclitaxeltreatment,MTT assay and BrdU assays were used to evaluate the cell proliferation of MCF-7cells.Results The expression of miR-200a in MCF-7cells increased after paclitaxel treatment in a dose-dependent manner,while the mRNA and protein expression of TFAMdecreased after paclitaxel treatment in a dose-dependent manner.MiR-200a mimics enhanced the inhibitory effect of paclitaxel on theproliferation of MCF-7cells,whereas anti-miR-200a attenuated the inhibitory effect of paclitaxel on MCF-7cell proliferation;TFAM alsoattenuated the inhibitory effect of paclitaxel on MCF-7cell proliferation.Conclusion MiR-200a and TFAM are involved in regulating thechemosensitivity MCF-7cells after paclitaxel treatment.