MiR-186-5p通过靶向调控RAB2A在乳腺癌细胞阿霉素耐药性中的逆转作用机制  

Reverse effect of miR-186-5p on adriamycin resistance of breast cancer cells through targeting RAB2A and its mechanism

作　　者：孙玉国[1] 王照岩[2] 杨玉玲[2] 杨志一 尹崇高[3] 李洪利 SUN Yu-guo;WANG Zhao-yan;YANG Yu-ling;YANG Zhi-yi;YIN Chong-gao;LI Hong-li(Dept of Pharmacy,Affiliated Hospital of Weifang Medical University;Dept of Pathology;College of Nursing;Medical Research Center,Weifang Medical University,Weifang Shandong 261053,China)

机构地区：[1]潍坊医学院附属医院药学部,山东潍坊261053 [2]潍坊医学院病理学教研室,山东潍坊261053 [3]潍坊医学院护理学院,山东潍坊261053 [4]潍坊医学院医学研究实验中心,山东潍坊261053

出　　处：《中国药理学通报》2018年第12期1668-1673,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金青年基金资助项目(No 81702932,81641111);山东省自然科学基金资助项目(No ZR2015HL065)

摘　　要：目的探讨miRNA-186-5p在乳腺癌细胞阿霉素耐药性逆转作用中的分子机制。方法采用阿霉素诱导人乳腺癌细胞系MCF-7,建立阿霉素耐药性细胞株,命名为MCF-7/ADR;采用双荧光素酶实验检测miR-186-5p与RAB2A的靶向结合情况;采用qRT-PCR和Western blot分别检测细胞中miR-186-5p、RAB2A mRNA的表达及RAB2A蛋白的表达情况;特异性上调MCF-7/ADR中的miR-186-5p,下调MCF-7中的miR-186-5p后,采用CCK-8和Western blot分别检测乳腺癌细胞增殖情况以及RAB2A蛋白的表达;采用Western blot和CCK-8分别检测共转染后,细胞增殖情况的变化及PI3K/Akt信号通路的激活情况。结果双荧光素酶实验结果显示,RAB2A是miR-186-5p的直接靶点;qRT-PCR结果显示,在MCF-7/ADR细胞中miR-186-5p mRNA表达明显下调,而RAB2A mRNA表达没有明显变化,RAB2A的蛋白表达明显上调。特异性上调MCF-7/ADR中的miR-186-5p,其增殖能力明显下降,RAB2A蛋白的表达明显下调;下调MCF-7中的miR-186-5p,其增殖能力明显增加,RAB2A蛋白的表达明显上调。Western blot和CCK-8结果显示,过表达miR-186-5p使PI3K/Akt信号通路表达明显减弱,且上调RAB2A逆转了上调miR-186-5p所致的增殖能力的减弱。结论 miR-186-5p通过靶向调控RAB2A,经PI3K/Akt信号通路调控乳腺癌细胞的阿霉素耐药和增殖。Aim To investigate the reverse effect of miR-186-5p on adriamycin resistance of breast cancer cells and the molecular mechanism. Methods The cell lines called MCF-7/ADR were established, which was induced by adriamycin. The targeted combination relationship of miR-186-5p and RAB2A was confirmed using dual luciferase assay. The mRNA levels of miR-186-5p and RAB2A, the protein levels of RAB2A were respectively detected using qRT-PCR and Western blot. CCK-8 and Western blot were respectively employed to assess the cell proliferation and the expression of RAB2A in breast cancer cells after over-expression of miR-186-5p in MCF-7/ADR cells and suppression of miR-186-5p in MCF-7cells. The changes of cell proliferation and the activation of PI3K/Akt signaling pathway were respectively evaluated by CCK-8 and Western blot after co-transfection. Results The results of dual luciferase experiments showed that RAB2A was a direct target of miR-186-5p. The qRT-PCR and Western blot results revealed that the mRNA levels of miR-186-5p was obviously down-regulated, the mRNA levels of RAB2A had no change, and the protein levels of RAB2A was visibly up-regulated in MCF-7/ADR. The capacity of cell proliferation and the expression of RAB2A protein obviously decreased after over-expression of miR-186-5p in MCF-7/ADR cells. The capacity of cell proliferation and the expression of RAB2A protein visibly increased after suppression of miR-186-5p in MCF-7 cells. Western blot results illustrated that over-expression of miR-186-5p significantly reduced the expression of PI3K/Akt signaling pathway. CCK8 results indicated that over-expression of miR-186-5p generated the attenuation of the proliferation ability that was restored by increasing RAB2A. Conclusion MicroRNA-186-5p targets RAB2A to modulate adriamycin resistance and proliferation of breast cancer cells via the PI3K/Akt signaling pathway.

关 键 词：乳腺癌 miR-186-5p RAB2A 阿霉素耐药 增殖 PI3K/AKT 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学] R342.2[医药卫生—基础医学]