Role Of MicroRNA-184 In Transforming Growth Factor Beta2-induced Epithelial Mesenchymal Transition In Human Lens Epithelial Cells

Objective: The epithelial-mesenchymal transition of postoperative residual lens epithelial cells is the major cause of pathology for after-cataract, and its molecular mechanisms, especially the regulation mechanism of its gene expression are unknown. Studies have shown that transforming growth factor β2(TGF-β2) is the most effective factor during the process of epithelial cell transition, and plays an important role in the pathogenesis of after-cataract. MicroRNA(microRNAs) is a class of non-coding single-stranded RNA, which can regulate the expression of target gene’s transcription level or translation level, so microRNAs play an important role in cell proliferation, apoptosis, differentiation, tumorigenesis and metabolism and other vital processes. Currently, using microRNAs for regulation of EMT has become a hot research. This paper aims to study how miR-184 mediated TGF-β2-induced EMT in human lens epithelial cells and their molecular mechanisms, by using the TGF-β2 induces the EMT of human lens epithelial cells in vitro and applying the microRNA regulation technology. Therefore, blocking the biological process of TGF-β2-induced transition of epithelial-mesenchymal provides a new idea for after-cataract prevention and treatment.Methods : Firstly, different concentrations of TGF-β2 was used to treat the cultured human lens epithelial cells HLE-B3. Then immunofluorescence method and western blotting method was used to observe and analyze the change of epithelial cell marker E-cadherin, mesenchymal cell marker vimentin and other cell transition related protein in epithelial cells, and the affect of TGF-β2 on expression of miR-184 was detected by utilizing the the qRT-PCR method. What’s more, the research model of epithelial-mesenchymal transition in human Lens Epithelial Cells was established. Using lipofectamineTM 2000-mediated miRNA regulation technology to transfect the HLE-B3 cells, and using the qRT-PCR method the mi R-184 expression of the cells that was transfected by miR-184 mimic and miR-184 inhibitor were detected respectively. At last, the Confocal laser and western blotting methods were utilized for observing the affect of up-regulated and down-regulated miR-184 gene on TGF-β2-induced lens epithelial-mesenchymal differentiation and on the expression of target genes bin3 and protein.Results: The exogenous TGF-β2 can induce epithelial-mesenchymal transition in human lens epithelial cells. Using gene technology the miRNA can regulate TGF-β2 induced human lens epithelial cells expressing mi R-184 and the related proteins expression as well as the expression of target genes bin3.Conclusions: miR-184 plays a key role in TGF-β2-induced human epithelial-mesenchymal transition of lens epithelial cells. Inhibition the expression of miR-184 can inhibit TGF-β2-induced epithelial-mesenchymal transition of lens epithelial cells, of which the molecular mechanism may be the inhibition of bin3 gene expression at the level of transcription and Translation.