MiR-142 Promotes Corneal Neovascularization Formation Via Up-regulating Akt2

Objective:To investigate the effect of miR-142 on the expression of cells in vitro,analyze the relationship between AKT2 and miR-142,and explore the influence and related mechanisms of corneal neovascularization.Methods:Control group,negative group,overexpression miR-142 group and knockdown miR-142 group were arranged.Mimic and inhibitor of miR-142 were used to overexpress or knock down miR-142,respectively.CCK-8 proliferation assay was performed to evaluate the impaction of miR-142 on proliferative ability of HUVECs.Dual-luciferase reporter gene system was used to verified that whether AKT2 is the target of miR-142.RT-PCR and Western blot were used to detect effect of miR-142 on AKT2.Results:Results of CCK8 assay showed that optical density(OD)of control group,negative group and overexpression miR-142 group was 872.21±44.20,842.16±49.54,1407.91±16.58,and the results had statistical significances(F=102.8,P=0.000).Knockdown miR-142 significantly weakened the proliferation rate of HUVECs(control group=919.69±25.46,negative group=943.43±40.53,knockdown miR-142 group=602.77±25.62,F=72.8,P=0.000).Consequence of dual-luciferase report gene system indicated that AKT2 wasn't the target of miR-142(control group=0.36,overexpressed miR-142 group=0.46,P=0.147).Q-PCR assay revealed that expression level of AKT2 was upregulated by overexpressing miR-142(0.408±0.046)than negative group(0.173±0.022)(P=0.018);inhibition of miR-142 blocked the expression of AKT2(negative group=0.362±0.068,down-regulated miR-142 group=0.160±0.019,P=0.037).Western blot assay verified the regulation of miR-142 on AKT2.Further study showed that the facilitation of miR-142 on proliferation of HUVECs was thoroughly blocked by shRNA-AKT2,results of control,overexpressed miR-142,overexpressed miR-142+shRNA-AKT2 and shRNA-142 were 981.80±75.40,1357.81±47.56,806.39±49.70,921.59±40.38,respectively,and the results had statistical significances(F=40.97,P=0.000).Conclusions:miR-142 facilitates the proliferation of HUEVCs via up-regulating AKT2,suggesting that miR-142 could promote the formation of corneal neovascularization.