MiR-182对乳腺癌细胞顺铂耐药的相关性研究  

Relationship Between the Mir-182 and the Cisplatin-Resistance in Breast Cancer

作　　者：龙景培[1] 万芳[1] 周俊[1] 张峰[1] 陈欣[1] 王一刻[1] 

机构地区：[1]浙江大学医学院附属妇产科医院外科,杭州310006

出　　处：《中国现代应用药学》2016年第6期721-726,共6页Chinese Journal of Modern Applied Pharmacy

摘　　要：目的探讨miR-182与乳腺癌细胞顺铂耐药性的关系。方法 MTT法检测miR-182对顺铂杀伤乳腺癌细胞能力的影响。利用生物信息学、定量PCR及Western blot法验证miR-182是否能调节乳腺癌细胞BNIP3的表达。运用JC-1染色、Annexin V染色及Western blot法研究miR-182影响顺铂疗效的信号通路。结果 miR-182模拟物可减弱顺铂对MCF-7细胞的杀伤活性,而miR-182抑制剂则增强顺铂对MCF-7细胞的杀伤活性。定量PCR及Western blot实验表明miR-182的靶基因可能为BNIP3。miR-182抑制剂联合顺铂可引起MCF-7细胞线粒体膜电位显著下降并诱导caspase-3的活化和凋亡的发生,转染BNIP3 siRNA后miR-182抑制剂联合顺铂对MCF-7细胞的凋亡诱导效应显著降低。结论 Mi R-182在乳腺癌中通过下调BNIP3的表达影响顺铂对乳腺癌细胞的杀伤活性。OBJECTIVE To investigate the relationship between the miR-182 and the cisplatin-resistance in breast cancer. METHODS MTT assay was performed to evaluate the role of miR-182 in cisplatin-induced cell death in MCF-7 cells. Bioinformatics, quantitative PCR and Western blot analysis was performed to determine whether the gene of BNIP3 was regulated by miR-182. JC-1 staining, Annexin V staining and Western blot analysis were performed to study the pathway of cell death induced by miR-182 inhibitors plus cisplatin in MCF-7. RESULTS Mi R-182 mimics impaired the anti-tumor effect of cisplatin. In contrast, the anti-tumor effect of cisplatin was significantly enhanced when the miR-182 inhibitors were transfected into MCF-7 cells. The results of Western blot and RT-PCR indicated that the BNIP3 gene was the target of miR-182. Furthermore, the apoptosis caused by miR-182 inhibitors plus cisplatin was dependent on the decrease of mitochondrial membrane potential and the activation of caspase-3 in MCF-7 cells. CONCLUSION Mi R-182 influences the anti-tumor effect of cisplatin in breast cancer by reducing the expression of BNIP3.

关 键 词：miR-182 BNIP3 乳腺癌 MCF-7 顺铂 

分 类 号：R730.21[医药卫生—肿瘤]