miR-106b～25基因簇增强乳腺癌MCF-7肿瘤干细胞相关耐药性  

Overexpression of miR-106b-25 cluster induces chemoresistance through regulating cancer stem cell-like phenotype in MCF-7 cell line

作　　者：胡蕴慧[1,2] 盛梦瑶 张孝云[1] 李双静[1] 范冬梅[1] 熊冬生[1] 周圆[1] 

机构地区：[1]中国医学科学院北京协和医学院血液病医院(血液学研究所),实验血液学国家重点实验室,天津300020 [2]天津医科大学附属肿瘤医院乳腺三科,天津300060

出　　处：《中国药理学通报》2013年第8期1070-1075,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81170512)

摘　　要：目的探讨乳腺癌细胞中miR-106b～25基因簇过表达与肿瘤耐药形成及乳腺癌干细胞特性的关系。方法利用慢病毒表达载体,在乳腺癌细胞系MCF-7中建立稳定过表达miR-106b～25基因簇,建立MCF-7-mirC细胞系,观察细胞对化疗药物敏感性的改变。利用细胞毒性实验、耐药克隆形成实验、细胞衰老相关β-半乳糖苷酶染色对细胞的耐药状况进行评价;通过流式细胞术和实时定量PCR测定P-gp的表达;通过微球体(mammosphere)培养实验及对乙醛脱氢酶(ALDH1)活性的检测评价MCF-7-mirC肿瘤干细胞特性的变化。结果过表达miR-106b～25基因簇能帮助细胞更快脱离由阿霉素引起的细胞衰老状态,促进耐药克隆的生长。但是,miR-106b～25基因簇对P糖蛋白(P-gp)在mRNA和蛋白水平表达均无影响。对ALDH1的检测及mammo-sphere形成实验结果显示,过表达miR-106b～25基因簇能提高MCF-7细胞中肿瘤干细胞的比例。结论 miR-106b～25基因簇对乳腺癌MCF-7细胞耐药性的调控不依赖于P-gp表达,至少与促进肿瘤干细胞特性有关。Aim To determine the effects of miR-106b 25 cluster on chemoresistant and stem cell-like phe- notype of breast cancer cells. Methods MiR-106b 25 cluster was overexpressed in breast cancer cell line MCF-7 by lentivirus, then the response of the cells to adriamycin was examined by MTT, drug resistant-clone formation assay and senescence-related 15-gal assay. The expression level of P-glycoprotein （P-gp） was detected by real-time PCR and flow cytometry. Mammosphere formation assay and ALDH1 activity assay were performed to indicate the cancer stem cell-like characteristics of MCF-7 cells after miR-106b -25 cluster overexpression. Results MCF-7 cells overcame adriamycin -induced senescence and became drug resistant by overexpression of the miR-106b -25 cluster, while P-gp was not involved in this process. The percentage of ALDH1high cells was elevated and more mammospheres formed after overexpressing miR-106 -25 cluster. Conclusion MiR-106b -25 cluster renders MCF-7 chemoresistance at least partly through regulating breast cancer stem cell characteristics, which is inde- pendent of P-gp expression.

关 键 词：miR-106b^25 乳腺癌 肿瘤干细胞 肿瘤耐药 细胞衰老 P糖蛋白 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学] R394.2[医药卫生—基础医学]