Intermediate conductance Ca2+ activated K+ channels 3.1 regulates the proliferation of human umbilical vein endothelial cells induced by epidermal growth factor in vitro

doi:10.16098/j.issn.0529-1356.2016.02.010

Abstract

Abstract:

Objective To examine the effect of epidermal growth factor (EGF) on the expression of intermediate conductance Ca²⁺ activated K+ channels 3.1 (KCa 3.1) in human umbilical vein endothelial cells (HUVECs), and to investigate the roles of KCa 3.1 in the HUVECs proliferation induced by EGF in vitro. Methods In this present study, the best working concentration of EGF was decided by MTT assay. The expression of KCa 3.1 was tested by immunofluorescence staining and western blotting assay after EGF-treatment. After the HUVECs were treated by TRAM-34, a blocker ofKCa 3.1, the EGF-induced proliferation of HUVECs was determined by MTT assay. Flow cytometry was performed to check the cell cycle of HUVECs. The mRNA expressions of cyclinD1 and cyclin-dependent kinase 4 (CDK4) were investigated by RT-PCR assay. Results After the HUVECs were treated with EGF for 48h, MTT assay showed the maximal stimulation of EGF at 25μg/L on the cell proliferation. EGF increased the expression of KCa 3.1 in immunofluorescence staining, and a significant 1.4-fold increase in KCa3.1-protein levels was revealed by Western blotting. Further research showed，after KCa 3.1 channels were selectively blocked with TRAM-34 for 48hours, the EGF-stimulated proliferation of HUVECs was suppressed observably in a dose-and time-dependent fashion, and the percentage of cells in G1 phase significantly increased in cell cycle. Interestingly, the blocked-KCa 3.1 down-regulated the expression of CDK4 in mRNA levels, whereas the cyclinD1 expression remained unchanged. Conclusions KCa 3.1 could potentially regulate the cell proliferations induced by EGF through modulating the cell cycle process in HUVECs.

GUO Hong-yu ZHAO Yu-jiao GAO Bo LI Xiao-dong ZHANG Ya-fang YANG Hui-ke*. Intermediate conductance Ca2+ activated K+ channels 3.1 regulates the proliferation of human umbilical vein endothelial cells induced by epidermal growth factor in vitro[J]. AAS, 2016, (2): 209-215.

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