Study on Effects of NHERF-1 on Proliferation and Migration of Human Umbilical Vein Endothelial Cells

摘要

Object To explore the effects of Na~+/H~+ exchanger regulatory factor 1 (NHERF1) on the phosphorylation levels of Akt1, activity of gelatinase secreted by HUVECs, and expression and distribution of cytoskeleton inside Human Umbilical Vein Endothelial Cells (HUVECs), and to expound the molecular mechanism of NHERF1 to influence the proliferation, migration and angiogenesis of vascular endothelial cells. Methods To construct the recombinant eukaryotic expression plasmid of NHERF1 and stably transfect HUVEC line with recombinant plasmid respectively; the 3-4,5-Dimethyl-2-thiazolyl-2,5-diphenyl-2H-tetrazolium bromide (MTT) was used to assay the proliferation activity of HUVECs after verification with the Western blotting method; to adopt the scarification test to measure the migration activity of HUVECs; to apply the Matrigel method to detect the angiogenesis ability of NHERF1 in cells; to detect the effect of NHERF1 on the phosphorylation levels of Akt1 by Western blotting; to use the gelatin zymography analysis to test the activity of gelatinase secreted by HUVECs; to observe the distribution of cytoskeleton with the immunofluorescence. Results The cDNA fragments of the exogenous NHERF-1 transfected have been integrated into the genome; NHERF-1 can significantly inhibit the proliferation, migration and angiogenesis of HUVECs, obviously down-regulate the phosphorylation levels of Akt11, make HUVECs decrease the secretion of reduction proenzyme and active enzyme, and also influence the distribution of cytoskeleton in vascular endothelial cells, compared to the cells in the control group. Conclusion NHERF1 can inhibit proliferation, migration and angiogenesis of HUVECs, and the inhibiting mechanism might relate to adjustment of the phosphorylation levels of Akt11, regulation of the activity of gelatinase secreted by HUVECs, and influence on the distribution of microfilament cytoskeleton.