miR-9在卵巢癌细胞上皮间质转化中的初步研究  

miR-9 Regulation of Epithelial-to-Mesenchymal Transition in Ovarian Cancer Cells

作　　者：周波[1] 孙朝阳[1] 夏萌[1] 卢运萍[1] 陈刚[1] 徐洪斌 

机构地区：[1]华中科技大学同济医学院附属同济医院肿瘤生物医学中心,湖北武汉430030 [2]深圳市人民医院妇科,广东深圳518020

出　　处：《现代生物医学进展》2016年第31期6001-6004,6009,共5页Progress in Modern Biomedicine

基　　金：国家自然科学基金项目(81272859);深圳科技计划项目(Jcyj20140416122811911;Jcyj20140416122811949)

摘　　要：目的:研究miR-9在卵巢癌细胞上皮间质转化(EMT)中的作用。方法:上调或者下调miR-9后,在RNA水平上通过RT-qPCR检测卵巢癌细胞系SKOV3和A2780中上皮指标E-cadherin表达变化;在蛋白水平,通过western blotting方法检测2株细胞系中上皮指标E-cadherin和间质指标vimentin蛋白表达变化。生物信息学预测可能靶向E-cadherin 3'UTR的miR NA,双荧光素酶报告系统进一步验证miR-9靶向结合E-cadherin的3'UTR区。结果:上调miR-9后,卵巢癌细胞系中E-cadherin表达受到明显抑制,vimentin表达明显增加;反之,下调miR-9后,E-cadherin表达明显增高,vimentin表达明显降低。通过生物信息学预测发现miR-9可以直接靶向E-cadherin的3'UTR区,荧光素酶报告系统验证预测结果正确。结论:miR-9促进卵巢癌细胞上皮间质转化。Objective： To study role of miR-9 in epithelial-to-mesenchymal transition（EMT） of ovarian cancer cells. Methods：RT-qPCR was used to detect the expression of E-cadherin after miR-9 mimics or inhibitor transfected in SKOV3 and A2780 cells at RNA level. Western blotting was performed to detect the expression of E-cadherin and vimentin at protein level. Bioinformatics analysis was conducted to predict the micro RNAs that may regulate the 3’ UTR of E-cadherin mRNA, which was further confirmed through Luciferase reporter assay. Results： E-cadherin was significantly decreased after miR-9 mimics transfected in SKOV3 and A2780 cells both at RNA and protein levels, while vimentin was markedly increased. However, these phenomena could be reversed after miR-9 inhibitor administration. The 3’ UTR of E-cadherin mRNA was predicted to be directly regulated by miR-9 via bioinformatics analysis. Luciferase reporter assay was performed to further confirm that miR-9 could directly target the 3’ UTR of E-cadherin mRNA. Conclusion： miR-9may promote EMT in ovarian cancer cells.

关 键 词：MIR-9 卵巢癌 侵袭与转移 E-CADHERIN 

分 类 号：R737.31[医药卫生—肿瘤]