卵巢癌细胞上皮间质转化及侵袭转移过程中microRNA-9的调控作用  

miR-9 Regulates Epithelial-to-Mesenchymal Transition,Invasion and Metastasis in Ovarian Cancer Cells

作　　者：孙朝阳[1] 李娜[1] 周波[1] 杨宗元[1] 卢运萍[1] 翁丹卉[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院妇产科,武汉430030

出　　处：《肿瘤防治研究》2014年第4期316-319,共4页Cancer Research on Prevention and Treatment

基　　金：国家青年基金资助项目(81000979/H1609)

摘　　要：目的探讨microRNA-9(miR-9)参与调控卵巢癌细胞EMT过程,以及影响卵巢癌细胞侵袭及转移的分子机制。方法使用TargetScan及PicTar数据库,预测可能靶向E-cadherin 3’UTR区的miRNA,双荧光素酶报告体系进行验证;上调候选miR后,用qRT-PCR和Western blot检测E-cadherin的表达变化;细胞免疫荧光观察E-cadherin、?-Catenin和Vimentin的表达;划痕实验、Transwell实验,观察卵巢癌细胞运动和侵袭能力的改变。结果 TargetScan、PicTar预测发现miR-9是唯一可与CDH1结合的miRNA。双荧光素酶报告系统验证预测结果正确。在SKOV-3中上调miR-9后,E-cadherin的表达受到显著抑制;细胞形态向间质细胞样转变,发生EMT分子水平的特征性改变;体外实验表明,卵巢癌细胞的运动和侵袭能力得到明显促进。结论 miR-9可以通过靶向调控E-cadherin表达,促进卵巢癌细胞的EMT进程,对卵巢癌细胞的运动和侵袭能力产生重要影响。Objective To determine the potential role of miRNAs in controlling EMT and metastasis in ovarian cancer cells. Methods Firstly, we used 2 computational algorithms, TargetScan and PicTar, to search for miRNAs that could target 3' UTR of CDH1 mRNA. Luciferase reporter assay, qRT- PCR and Western blot were carried out to validate the results. Immunofl uorescence staining of E-cadherin, ?-Catenin and Vimentin were performed to detect the EMT process of ovarian cancer cell lines transfected with miRNAs. Wound healing assay and Transwell assay were used to detect the cell motility and invasiveness. Results miR-9 directly regulated CDH1, causing a remarkable inhibition of E-cadherin expression. miR-9 also caused a dramatic increasing of ?-Catenin and Vimentin expression, which were the important molecular markers of EMT process. Moreover, up-regulation of miR-9 in ovarian cancer cell lines signifi cantly improved cell motility and invasiveness. Conclusion miR-9 has a significantly promoting effect on EMT via directly targeting E-cadherin in ovarian cancer cells, and may play a potential important role in cancer metastasis and invasiveness.

关 键 词：microRNA-9 上皮间质转化(EMT) E-钙粘蛋白 卵巢癌 

分 类 号：R737.31[医药卫生—肿瘤]