Med19基因敲减增加乳腺癌细胞化疗敏感性及其可能的机制

发布时间：2018-05-04 06:54

本文选题：中介体 + 乳腺癌　；参考：《中国肿瘤生物治疗杂志》2017年04期

【摘要】：目的:研究中介体(mediator,Med)19对乳腺癌化疗敏感性的影响并分析其分子机制。方法:选用多柔比星(adriamycin,ADM)耐药的人乳腺癌细胞MCF-7/ADM和亲本细胞MCF-7(NC组),采用慢病毒载体介导RNA干扰方法构建Med19稳定低表达的MCF-7/ADM与MCF-7细胞株(KD组),并用Real-time PCR和Western blotting方法验证干扰效果。CCK-8法检测慢病毒介导的Med19敲减前后两种细胞对ADM、顺铂(cisplatin,DDP)和紫杉醇(taxinol,TAX)药物敏感性的变化。Real-time PCR和Western blotting检测Med19敲减对多药耐药基因1(multidrug resistance 1,MDR1)和细胞凋亡基因Bcl2、Bax及Caspase-3、active Caspase-3的表达的影响。流式细胞术检测敲减Med19及ADM处理对细胞凋亡的影响。结果:与MCF-7相比,MCF-7/ADM细胞对ADM、DDP和TAX均具有耐药性。成功构建Med19稳定低表达的MCF-7/ADM与MCF-7细胞株,并且其对ADM、DDP、TAX的敏感性增加,药物作用IC_(50)显著降低(均P0.05)。MCF-7/ADM细胞Med19 mRNA和蛋白表达显著高于MCF-7细胞,Med19的敲减可降低MCF-7/ADM细胞中MDR1 mRNA与蛋白表达水平(均P0.05)并可增加MCF-7/ADM及MCF-7细胞中凋亡相关active Caspase-3、Bax的蛋白表达,降低Bcl2的蛋白表达(均P0.05)。此外,与NC及NC+ADM组相比,KD组及KD+ADM组凋亡水平显著增加(均P0.05)。结论:Med19高表达参与乳腺癌化疗耐药,其机制可能与Med19调节MDR1的表达并影响细胞凋亡有关。
[Abstract]:Aim: to study the effect of mediatorus Medan19 on the chemosensitivity of breast cancer and analyze its molecular mechanism. Methods: human breast cancer cell line MCF-7/ADM and parent cell line MCF-7(NC were used to construct stable and low expression MCF-7/ADM and MCF-7 cell line of Med19 by RNA interference mediated by lentivirus vector. Real-time PCR and Western blotting methods were used to test the results. The effect of CCK-8 on the sensitivity of Lentivirus mediated Med19 knockout to Adm, Cisplatin cisplatin (DDP) and taxinolus (TAX). Real-time PCR and Western blotting Detection of Med19 knock down multidrug Resistance Gene 1(multidrug resistance 1 MDR1) and apoptosis Gene Bcl2Bax And the expression of Caspase-3 active Caspase-3. The effects of knockout Med19 and ADM on apoptosis were detected by flow cytometry. Results: compared with MCF-7, MCF-7 / ADM cells were resistant to ADMN DDP and TAX. MCF-7/ADM and MCF-7 cell lines with stable and low expression of Med19 were successfully constructed, and their sensitivity to ADMS-DDPTAX was increased. The expression of Med19 mRNA and protein in P0.05).MCF-7/ADM cells was significantly higher than that in MCF-7 cell line Med19 (P 0.05) and the protein expression of active Caspase-3 was increased in MCF-7/ADM and MCF-7 cells. The protein expression of Bcl2 was decreased (P 0.05). In addition, compared with NC and NC ADM groups, apoptosis levels in KD and KD ADM groups were significantly increased (P 0.05). ConclusionThe high expression of w / Med19 may be involved in the chemotherapeutic resistance of breast cancer, and its mechanism may be related to the regulation of MDR1 expression by Med19 and its effect on apoptosis.
【作者单位】：江南大学附属医院肿瘤研究所;江南大学附属医院肿瘤外科;
【基金】：国家自然科学基金面上项目资助(No.81472485) 无锡市卫计委资助项目(No.Q201615,YGZXM14038No.YGZXQ1311)~~
【分类号】：R730.5;;R737.9

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