miR-142 promotes corneal neovascularization formation via up-regulating AKT2

Abstract:

Objective：To study the impact of miR-142 on corneal neovascularization formation and related mechanism. Methods：Con－trol 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 im－paction of miR-142 on proliferative ability of human umbilical vein endothelial cells（HUVECs）. Dual-luciferase reporter gene system was used to verify 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 absorbance（A） of control group，negative group and overexpression miR-142 group was 872.21±44.20，842.16±49.54，1 407.91±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）. RT-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，overex－pressed miR-142，overexpressed miR-142+shRNA-AKT2 and shRNA-142 were 981.80±75.40，1 357.81±47.56，806.39±49.70，921.59±40.38，respectively，and the results had statistical significances（F=40.97，P=0.000）. Conclusion：miR-142 facilitates the proliferation of HUEVCs via up-regulating AKT2，suggesting that miR-142 could promote the formation of corneal neovascularization.