微小RNA mir-145在乳腺癌细胞中调控Nanog的机制与功能研究  

Mechanism and Function of mir-145 on Regulating Nanog in Breast Cancer Cells

作　　者：骆启聪[1] 张环[1] 陈蓉珊 陈刚[1] 米彦军[1] LUO Qicong ZHANG Huan CHEN Rongshan CHEN Gang MI Yanjun(The First Affiliated Hospital of Xiamen University, Xiamen Fujian 361003, China)

机构地区：[1]厦门大学附属第一医院,福建厦门361003

出　　处：《长沙大学学报》2016年第5期10-13,16,共5页Journal of Changsha University

基　　金：厦门科技惠民计划项目(批准号:3502Z20134005)

摘　　要：目的:探讨mir-145在乳腺癌细胞中对Nanog的调控作用及功能.方法:采用荧光定量PCR检测乳腺癌细胞系中mir-145与Nanog表达水平,利用流式细胞术比较不同乳腺癌细胞系中肿瘤干细胞比例,悬浮培养瘤球并比较瘤球与贴壁细胞间mir-145、Nanog的表达差异,过表达mir-145后检测Nanog表达水平与报告基因活性,流式细胞术检测mir-145表达升高后乳腺癌干细胞比例,平板克隆形成实验检测mir-145表达升高后乳腺癌细胞成瘤能力.结果:T-47D细胞表达高水平的mir-145和低水平的Nanog,MDA-MB231表达低水平的mir-145和高水平的Nanog,MDA-MB231细胞乳腺癌干细胞比例高于T-47D细胞,瘤球细胞中的mir-145表达水平低于贴壁细胞,而Nanog表达水平高于贴壁细胞.过表达mir145降低Nanog表达水平与报告基因活性,降低乳腺癌干细胞比例与平板克隆形成能力.结论:mir-145能够调控Nanog表达水平,影响乳腺癌细胞干细胞特性.Objective： To investigate the mechanism and function of mir- 145 on regulating Nanog in breast cancer cells. Methods：Comparing the expression levels of mir- 145 and Nanog of breast cancer cells using Real- Time PCR,comparing the population of cancer stem cells via FACS assay,and culturing and detecting the expression level of mir- 145 and Nanog in breast cancer sphere.Breast cancer cells were transfected with mir- 145 mimic. Fluorescent quantitative PCR was used to determine mir- 145 and Nanog mRNA expression and dual reporter assay was used to determine Nanog protein expression. Results： T- 47 D cell expressed higher level of mir- 145,lower level of Nanog,and smaller population of breast cancer stem cells,while MDA- MB231 cell expressed lower level of mir- 145,higher level of Nanog,and larger population of breast cancer stem cells. The mir- 145 expression level was lower in sphere and Nanog was higher in sphere comparing to adherent cells. The mir- 145 overexpression in breast cancer cells inhibited Nanog mRNA and protein expression and reduced the stem cell population and colony- formation ability. Conclusion： mir- 145 can regulate Nanog expression and reverse stemness of breast cencer cells.

关 键 词：乳腺癌 NANOG MIR-145 肿瘤干细胞 

分 类 号：Q756[生物学—分子生物学]