中国肿瘤生物治疗杂志2017，Vol.24Issue(4)：380-388,9.DOI:10.3872/j.issn.1007-385X.2017.04.008
Med19基因敲减增加乳腺癌细胞化疗敏感性及其可能的机制
Med19 knockdown promotes chemosensitivity of breast cancer cells and its possible mechanism
摘要
Abstract
Objective:To explore the effect and molecular mechanism of mediator 19 (Med19) on drug sensitivity of breast cancer cells.Methods:Human breast cancer cell line MCF-7/ADM (adriamycin resistant) and it's parental cell line MCF-7 (NC groups) were selected.Lentivirus-mediated RNA-interference was used to construct MCF-7/ADM and MCF-7 cell lines that stably expressing low Med19 (KD groups),and the infection efficiency was detected by Real-time PCR and Western blotting assays.The change of drug-susceptibility to ADM,cisplatin (DDP) and taxinol (TAX) were examined with CCK-8 assay before and after Med19 knockdown.The effects of Med19 knockdown on the expressions of multidrug resistance gene 1 (MDR1),Bcl2,Bax,Caspase-3 and active caspase-3 in breast cancer cells were detected by RT-PCR and Western Blotting assays,respectively.Effect of Med19 down-regulation and ADM treatment on apoptosis rates of MCF-7 and MCF-7/ADM cells were detected with flow cytometry assay.Results:Compared with MCF-7 cells,MCF-7/ADM cells exhibited drug resistance to ADM,DDP and TAX.MCF-7/ADM and MCF-7 cell lines that stably expressing low Med19 level were successfully established;their drug-susceptibilities to ADM,DDP,TAX were increased,and the IC50 values of these three cytotoxic agents deceased obviously (all P ＜ 0.05).The mRNA and protein expressions of Med19 in MCF-7/ADM cells were significantly higher than those in MCF-7 cells,however,Med19 knockdown significantly inhibited the expressions of MDR1 mRNA and protein in MCF-7/ADM cells (all P ＜ 0.05);in addition,it significantly increased the expressions of active caspase-3 and Bax protein in both MCF-7/ADM and MCF-7 cells (all P ＜ 0.05)and decreased the expression of Bcl2 protein (all P ＜ 0.05).Besides,compared with NC or NC + ADM group,the apoptosis rate of the cells significantly increased in KD group or KD + ADM group (all P ＜ 0.05).Conclusions:High Med19 expression plays a particularly important role in chemoresistance of breast cancer,and the mechanism might correlate with Med19 regulating MDR1 expression and affecting apoptosis related genes.关键词
中介体19/乳腺癌/多药耐药/多柔比星Key words
mediator19/breast cancer/multidrug resistance/adriamycin分类
医药卫生引用本文复制引用
刘贝贝,张秀芬,高丹凤,郭子健,李莉华..Med19基因敲减增加乳腺癌细胞化疗敏感性及其可能的机制[J].中国肿瘤生物治疗杂志,2017,24(4):380-388,9.基金项目
国家自然科学基金面上项目资助(No.81472485) （No.81472485）
无锡市卫计委资助项目(No.Q201615,YGZXM14038No.YGZXQ1311).Project supported by the National Natural Science Foundation of China(No.81472485),and the Project of the Wuxi Health and Family Planning Commission (No.Q201615,YGZXM14038,YGZXQ1311) （No.Q201615,YGZXM14038No.YGZXQ1311）
