Up-regulation of SET Protein in the Proliferation of GC-2 spd Cells and Histone Acetylation

ZHANG Bei; ZHU Qian; XU Bo-qun; XU Wen-dan; YANG Xiao-yu; GAO Chao; GAO Li; LIU Jia-yin; CUI Yu-gui

doi:10.3969/j.issn.1674-1889.2017.02.002

Abstract：Objective:To investigate the effects of SET protein on the regulation of spermiogenesis,the proliferation and histone acetylation of spermatocyte cell line,GC-2 spd,were observed.Methods:The GC-2 spd cell was cultured in vitro,and identified by two markers of spermatocytes,lactate dehydrogenase C (LDHC) and Testis-Specific Kinase 1 (TESK1).The combination of SET and H4 was detected by an immunoprecipitation kit.The cells were transfected with AdH1-siRNA/SET to knockdown the expression of SET protein,AdH1-siRNA/NS as control.The cell proliferation was compared between two groups.The expressions of SET protein,histone acetyltransferases and histone deacetylases were detected using realtime RT-PCR and Western Blot.Results:Two markers of spermatocytes,LDHC and TESK1,were positively expressed in the GC-2 spd cell.SET protein was expressed in both cytoplasm and nucleus of GC-2 spd cell.Immunoprecipitation showed that SET protein combined with H4.The expression of SET protein in the cytoplasm and nucleus of the cells transfected cells with AdH1siRNA/SET was significantly decreased (P＜0.01),while the cell proliferation significantly inhibited.Interestingly,the acetylation level of H4 was significantly increased in the transfected cells (P＜0.05).However,there were no significantly differences in the expressions of HATs and HDACs (P＞0.05).Conclusions:SET protein was expressed in both cytoplasm and nucleus of spermatocytes,the knockdown expression of SET protein inhibited the cell proliferation.SET protein participated in the regulation of spermiogenesis,probably by regulating the acetylation level of histone H4.