【摘 要】
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High glucose affects cell-cycle progression in many cells.The aim of this study was to explore whether high glucose-induced Erk1/2 activation regulates cell
【机 构】
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Key Laboratory of Fertility Preservation and Maintenance of Ministry of Education,Department of anot
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High glucose affects cell-cycle progression in many cells.The aim of this study was to explore whether high glucose-induced Erk1/2 activation regulates cell cycle-related proteins in human ovarian granulose cell-like KGN cells.In the present study, KGN cells were treated with high concentrations glucose (25 mM) with or without addition of Erk 1/2 inhibitor PD98059(25 μM) or a specific siRNA for Erk 1/2.Cell cycle was analyzed by flow cytometry, The expression level of Erk 1/2, cyclin D1 and P21 in KGN cells was determined by real-time PCR, immunohistochemistry and western blotting.High glucose stimulates can induce G1 phase arrest through ERK/MAPK signal pathway.High glucose stimulates phosphorylation of Erk 1/2 in KGN cells.Activation of Erk1/2 leads to phosphorylation of p27Kip1 serine(178) in vitro.Treatment with PD98059 significantly inhibited Erk 1/2 expression(p<0.05).The reduction of Erk1/2 activity led to an increase of cellular p27Kip1 and p21CIP1 levels in KGN cells and downregulation of cyclinD1 (p<0.05).We then observed that ERK1/2 levels decreased in the presence of specific siRNA targetingof KGN cells,and also upregulated p27Kipl and p21CIP1 and downregulated cyclinD1(p<0.05).The results indicate that activation of Erk1/2 in human granulosa cells is responsible for high glucose stimulation.High Erk1/2 activation regulated the cell cycle-related proteins.
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