miR-9 在卵巢癌细胞上皮间质转化中的初步研究

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。

miR-9 Regulation of Epithelial-to-Mesenchymal Transition in OvarianCancer Cells

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 atRNA level. Western blotting was performed to detect the expression of E-cadherin and vimentin at protein level. Bioinformatics analysiswas conducted to predict the microRNAs that may regulate the 3'' UTR of E-cadherin mRNA, which was further confirmed through Luciferasereporter assay.Results:E-cadherin was significantly decreased after miR-9 mimics transfected in SKOV3 and A2780 cells bothat RNA and protein levels, while vimentin was markedly increased. However, these phenomena could be reversed after miR-9 inhibitoradministration. The 3'' UTR of E-cadherin mRNA was predicted to be directly regulated by miR-9 via bioinformatics analysis. Luciferasereporter 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.