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

关键词:  miR-182  BNIP3  乳腺癌  MCF-7  顺铂

Key words:  MiR-182  BNIP3  breast cancer  MCF-7  cisplatin

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 MiR-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。

Abstract:

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 MiR-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 MiR-182 influences the anti-tumor effect of cisplatin in breast cancer by reducing the expression of BNIP3.