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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 miR-182 and Cisplatin resistance in breast cancer cells. Methods The effect of miR-182 on the ability of cisplatin to kill breast cancer cells was detected by MTT assay. Bioinformatics, quantitative PCR and Western blot were used to verify whether miR-182 regulates BNIP3 expression in breast cancer cells. JC-1 staining, Annexin V staining and Western blot were used to investigate the effect of miR-182 on cisplatin-induced signal pathways. Results miR-182 mimics decreased the killing activity of cisplatin on MCF-7 cells, while miR-182 inhibitor enhanced the killing activity of cisplatin on MCF-7 cells. Quantitative PCR and Western blot showed that miR-182 target gene may be BNIP3. miR-182 inhibitor combined with cisplatin can induce a significant decrease of mitochondrial membrane potential and induce the activation and apoptosis of caspase-3 in MCF-7 cells. After transfection with BNIP3 siRNA, miR-182 inhibitor and cisplatin can inhibit MCF-7 cells The apoptosis-inducing effect was significantly reduced. Conclusions Mi R-182 inhibits the killing activity of cisplatin on breast cancer cells by down-regulating BNIP3 expression in breast cancer.