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Dan Wu, Jiaxu Hong, JianJiang Xu, Jianjiang,XU,ocular surface disease group in Shanghai Eye & ENT Hospital of Fudan University; MicroRNA-494 is Highly Expressed in Human Cornea Epithelium Cells(HCECs) and inhibits Nerve Growth Factor(NGF)-Induced Cell Proliferation through Targeting Cyclin D1. Invest. Ophthalmol. Vis. Sci. 2014;55(13):5517.
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© ARVO (1962-2015); The Authors (2016-present)
To reveal novel miRNA regulators implicated in NGF-induced HCECs proliferation.
The differential miRNA expression profiles in HCECs with or without 25ng/ml NGF treatment were identified by microarray analysis and confirmed by RT-PCR.The lentiviral vector contained object miRNA were transfected into HCECs.The expression level of NGF-related miRNA and its target gene were examined by RT-PCR,western blot.Furthermore, the cell cycle distribution was performed by flow cytometry.
Five miRNAs were up-regulated and three miRNAs down-regulated in NGF treated HCECs when compared with non-treated cells.The most obvious change was the apparent decrease of miR-494(P<0.05).Transfection of the lentivirus pLV-THM-miR-494(figure 1) greatly recovered miRNA-494 expression(P<0.05,figure 1).Cyclin D1 was predicted as a novel target of miRNA-494 through bioinformatics analysis.Overexpression of the miR-494 suppressed cyclin D1 protein and mRNA expression and increased cell distribution in G1 period(P<0.05,figure 2),which showed obviously suppressed proliferation ability.However,NGF treatment largely reversed the effects induced by miR-494, suggesting that the regulative effect of miR-494 on HECEs growth was indeed mediated by NGF(P<0.05,figure1 and figure 2)
MiRNA-494 and its downstream target Cyclin D1 could be a crucial axis for NGF in regulation the proliferation of HCECs.Specific modulation of miR-494 in human cornea epithelium cells may represent an attractive approach for the treatment of cornea epithelium defects diseases.
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