Journal of Integrative Agriculture ›› 2014, Vol. 13 ›› Issue (4): 827-836.DOI: 10.1016/S2095-3119(13)60430-X

• 论文 • 上一篇    下一篇

17β-Estradiol Regulates SKP2 Expression in Cultured Immature Boar Sertoli Cells Mainly via Estrogen Receptor β, cAMP-PKA and ERK1/2

 WANG Xian-zhong, ZHU Feng-wei, WANG Yong, WANG Yi, ZHANG Jiao-jiao , ZHANG Jia-hua   

  1. Chongqing Key Laboratory of Forage & Herbivore, College of Animal Science and Technology, Southwest University, Chongqing 400716, P.R.China
  • 收稿日期:2012-12-05 出版日期:2014-04-01 发布日期:2014-04-16
  • 通讯作者: ZHANG Jia-hua, E-mail: jhzhang007@aliyun.com
  • 作者简介:WANG Xian-zhong, Tel: +86-23-68250215, Fax: +86-23-68251196, E-mail: xianzhong_wang@aliyun.com;
  • 基金资助:

    This work was supported by grants from the National Natural Science Foundation of China (31072183) and the Fundamental Research Funds for the Central Universities, China (XDJK2009B035).

17β-Estradiol Regulates SKP2 Expression in Cultured Immature Boar Sertoli Cells Mainly via Estrogen Receptor β, cAMP-PKA and ERK1/2

 WANG Xian-zhong, ZHU Feng-wei, WANG Yong, WANG Yi, ZHANG Jiao-jiao , ZHANG Jia-hua   

  1. Chongqing Key Laboratory of Forage & Herbivore, College of Animal Science and Technology, Southwest University, Chongqing 400716, P.R.China
  • Received:2012-12-05 Online:2014-04-01 Published:2014-04-16
  • Contact: ZHANG Jia-hua, E-mail: jhzhang007@aliyun.com
  • About author:WANG Xian-zhong, Tel: +86-23-68250215, Fax: +86-23-68251196, E-mail: xianzhong_wang@aliyun.com;
  • Supported by:

    This work was supported by grants from the National Natural Science Foundation of China (31072183) and the Fundamental Research Funds for the Central Universities, China (XDJK2009B035).

PDF

摘要： Estrogen plays an important role in regulating testicular Sertoli cell number. Furthermore, S-phase kinase-associated protein 2 (SKP2) plays a central role in mammalian cell cycle progression. The objective of this study was to determine whether 17β-estradiol can regulate the expression of SKP2, and the Sertoli cell cycle, via estrogen receptor β (ERβ), the cyclic adenosine monophosphate (cAMP)-protein kinase A (PKA) and extracellular signal-regulated kinase (ERK1/2) pathway. When cultured immature boar Sertoli cells were treated with 17β-estradiol, a time-dependent increase in SKP2 mRNA and protein level was observed by real-time PCR and Western blot, and 17β-estradiol activity peaked at 30 min. Treatment with ICI182780 and ERβ antagonist reduced 17β-estradiol-induced expression of SKP2 and proliferating cell nuclear antigen (PCNA), while increasing the protein concentration of p27kip1. However, the effect of ERa antagonist on these parameters was lower than that of ICI182780 and ERβ. Forskolin had a similar effect as 17β-estradiol on the expression of SKP2, PCNA and p27kip1. Rp-cAMP, H-89 and U0126 treatment reduced 17β-estradiol-induced changes, while H-89 also inhibited ERK1/2 activation. Therefore, 17β-estradiol mainly regulates SKP2 mRNA and protein expression via ERβ-cAMP-PKA and ERK1/2 activation. SKP2 and PCNA expression were positively correlated, while increased SKP2 expression likely resulted in p27kip1 degradation.

关键词: 17&beta, -estradiol , Sertoli cell , SKP2 , estrogen receptor , cAMP-PKA , ERK1/2

Abstract: Estrogen plays an important role in regulating testicular Sertoli cell number. Furthermore, S-phase kinase-associated protein 2 (SKP2) plays a central role in mammalian cell cycle progression. The objective of this study was to determine whether 17β-estradiol can regulate the expression of SKP2, and the Sertoli cell cycle, via estrogen receptor β (ERβ), the cyclic adenosine monophosphate (cAMP)-protein kinase A (PKA) and extracellular signal-regulated kinase (ERK1/2) pathway. When cultured immature boar Sertoli cells were treated with 17β-estradiol, a time-dependent increase in SKP2 mRNA and protein level was observed by real-time PCR and Western blot, and 17β-estradiol activity peaked at 30 min. Treatment with ICI182780 and ERβ antagonist reduced 17β-estradiol-induced expression of SKP2 and proliferating cell nuclear antigen (PCNA), while increasing the protein concentration of p27kip1. However, the effect of ERa antagonist on these parameters was lower than that of ICI182780 and ERβ. Forskolin had a similar effect as 17β-estradiol on the expression of SKP2, PCNA and p27kip1. Rp-cAMP, H-89 and U0126 treatment reduced 17β-estradiol-induced changes, while H-89 also inhibited ERK1/2 activation. Therefore, 17β-estradiol mainly regulates SKP2 mRNA and protein expression via ERβ-cAMP-PKA and ERK1/2 activation. SKP2 and PCNA expression were positively correlated, while increased SKP2 expression likely resulted in p27kip1 degradation.

Key words: 17β-estradiol , Sertoli cell , SKP2 , estrogen receptor , cAMP-PKA , ERK1/2