miR-26b调控MCF-7细胞MCL-1蛋白的表达并诱导凋亡  

miR-26b modulating the expression of MCL-1 protein and the inducing cell apoptosis in MCF-7 cells

作　　者：林峰[1] 陈志杭 

机构地区：[1]杭州市余杭区妇幼保健院,浙江杭州310000 [2]杭州市西湖区古荡街道社区卫生服务中心,浙江杭州310013

出　　处：《中国卫生检验杂志》2015年第19期3327-3329,3332,共4页Chinese Journal of Health Laboratory Technology

摘　　要：目的探讨microRNA-26b(miR-26b)的作用靶点及其对MCF-7细胞的抑制作用。方法通过荧光定量PCR方法及Western blot方法检测正常乳腺细胞系MCF-10A及乳腺癌细胞系MCF-7、MDA-MB-231及MDAMB-435的miR-26b及MCL-1的表达水平。MTT法检测miR-26b转染对MCF-7细胞活力的影响,Annexin V/PI染色法检测miR-26b转染对MCF-7细胞凋亡的影响。通过生物信息学分析预测miR-26b的靶基因,并将miR-26b转染入MCF-7细胞,检测miR-26b对MCL-1表达水平的影响。结果相比于MCF-10A细胞,乳腺癌细胞系MCF-7、MDA-MB-231及MDA-MB-435的miR-26b表达水平均显著下降,同时MCL-1表达水平均显著上升,提示miR-26在乳腺癌中其肿瘤抑制作用可能调节细胞MCL-1的表达。进一步研究发现MCL-1基因的3'-UTR区存在miR-26b的假定结合位点,且在乳腺癌细胞中转染miR-26b后,MCL-1的表达下降。转染miR-26b可抑制MCF-7细胞的活力并诱导其发生凋亡。结论 miR-26b下调MCF-7细胞MCL-1蛋白的表达并诱导其发生凋亡。Objective To investigate the target of miR- 26 b and the viability inhibition to MCF- 7 cells. Methods Expressions of miR- 26 b and MCL- 1 were detected using RT- q PCR and Western blot in normal breast cell line MCF- 10 A and the breast cancer cell lines MCF- 7,MDA- MB- 231 and MDA- MB- 435. The cell viability was evaluated by MTT assay after the MCF- 7 cells were transfected with miR- 26 b,and the effect of miR- 26 b on the apoptosis was detected by Annexin V / PI staining. Predict the target gene of miR- 26 b using bioinformatic analysis. Transfect miR- 26 b into the MCF- 7 cells,and then detect the effect of miR- 26 b on the level of MCL- 1. Results The expression of miR- 26 b was down- regulated and MCL- 1 expression level was up- regulated in breast cancer cell lines MCF- 7,MDA- MB- 231 and MDA- MB- 435,compared with the MCF- 10 A,suggesting the miR- 26 b may act as a tumor suppressor and regulating the expression of MCL-1 in breast cancer. Bioinformatic analysis identified a putative target site in the MCL- 1 gene 3＇- UTR for miR- 26 b,and after transfection of miR- 26 b,the MCL- 1 expression in MCF- 7 cells decreased. Transfection of miR- 26 b can inhibit the cell viability in MCF- 7 cells,and induce cell apoptosis. Conclusion MiR- 26 b down- regulates the expression of MCL- 1 and induces cell apoptosis in MCF- 7.

关 键 词：miR-26b MCL-1 乳腺癌 凋亡 

分 类 号：R737.9[医药卫生—肿瘤]