The influence of microRNA-184 transfection on the biological behavior of human umbilical vein endothelial cells in vitro / 中华实验眼科杂志

ABSTRACT

Background Previous studies showed that microRNA-184 (miR-184) might inhibit the biological function of vascular endothelial cells.In ophthalmology,to determine the inhibitory effect of miR-184 on angiogenesis is of clinical significance for the prevention and treatment of corneal inflammation,retinal and choroidal neovascularization.Objective This study was to verify the influence of miR-184 transfection on the proliferation,migration and angiogenic ability of HUVECs and explore its mechanism.Methods HUVECs line was cultured in vitro and divided into 4 groups.No transfection regent and RNA molecule were added in the medium of the blank control group;only transfection regent was added in the blank transfection group; hsa-miR-184 mimic/negative was added in the negative control group,and Cy3 labeled hsa-miR-184 mimic-lipofectamine 2 000 mixture was added in the miR-184 transfection group.The proliferation of HUVECs (absorbance,A490) was detected by MTT.The transmembrane cell number was counted by Transwell insert to evaluate the migration ability of HUVECs.A three dimensional cultural system of cells was constructed on the matrigel,and tube formation number of HUVECs was assessed.Results Positive HUVECs for Cy3 labeled siR-RiboTM + hposome showed red fluorescence,with the optimal transfecting concentration ＞ 100 nmol/L.The relative expression levels of miR-184 were 1 524.10±385.89 and 1.00±0.05 in the miR-184 transfection group and the negative control group,showing a significant difference (t=-7.894,P＜0.01).The A490value of HUVECs was 0.50±0.04 and the number of migrating cells through the transwell membrane was (55.40±5.86)/field in the miR-184 transfection group,and they are significantly reduced in comparison with (0.62±0.04) and (83.40±5.59)/field in the negative control group (t =5.639,7.730,all at P＜ 0.01).A significant difference was found in the total branch points of capillary tubes between the miR-184 transfection group and negative control group ([13.33 ± 2.08]/field versus [22.00 ± 2.00]/field) (t =5.511,P＜ 0.05).Conclusions MiR-184 can suppress the proliferation,migration and tubing of HUVECs after transfection in vitro.MiR-184 participates in the regulation of angiogenesis.