Cell Biology and Toxicology

, Volume 26, Issue 3, pp 265–277 | Cite as

Low-dose ethanol suppresses 17β-estradiol activity in GH4C1 pituitary tumor cells

  • Guoxin Wang
  • Dawei Chen
  • Haoshu Luo
  • Jiali Liu
  • Xiaowen Ji
  • Jingjing Fan
  • Sheng Cui
Article

Abstract

The interaction of chronic alcohol consumption with estrogen has been recognized in various tissues; however, the potential mechanism has not been clearly defined. In this study, we made the following observations: (a) 0.01% (v/v) ethanol (corresponding to 1.7 mM) significantly elevated estrogen receptor α (ERα) protein content, stimulated activator protein-1 (AP-1)-dependent ERα transcriptional activities, and ultimately enhanced GH4C1 cells growth in vitro and (b) the same concentration of ethanol suppressed the stimulatory effects of 17β-estradiol (E2; 10 nM) on both cell growth and cellular PRL accumulate through attenuation of ERα actions at both the estrogen response element and the AP-1 site. These observations raise the question as to what extent ethanol influences signal transduction pathways controlled by E2 in experimental medicine and biology.

Keywords

AP-1 ERα Ethanol GH4C1 cells Pituitary 17β-estradiol 

Abbreviations

AP-1

activator protein-1

BrdU

bromodeoxyuridine

EGF

epidermal growth factor

ER

estrogen receptor

ERE

estrogen response element

EtOH

ethanol

E2

17β-estradiol

GAPDH

glyceraldehyde-3-phosphate dehydrogenase

GFP

green fluorescent protein

GH4

GH4C1

HPRT

hypoxanthine phosphoribosyltransferase

IGF-1

insulin-like growth factor 1

MTT

methylthiazolyldiphenyl-tetrazolium bromide

PCR

polymerase chain reaction

PMSF

phenylmethanesulfonyl fluoride

PRL

prolactin

RNAi

RNA interference

shRNA

short hairpin RNA

SOE

splicing by overlap extension

TRITC

tetramethyl rhodamine isothiocyanate

Notes

Acknowledgments

This study was supported by grants to Sheng Cui from the Natural Science Foundation of China (30630011). We thank Dr. Weidong Han (Department of Molecular Biology, Institute of Basic Medicine, Chinese PLA General Hospital, Beijing) for kindly providing pSG5-hERα plasmid. The pERE-TK-Luc and pAP1-Luc reporter plasmid were gifts from Prof. Dalong Ma (Chinese National Human Genome Center, Beijing). We also thank Prof. Beate Brand-Saberi (Department of Molecular Embryology, Institute of Anatomy and Cell Biology, Freiburg University, Germany) for the generous gift of the peGFP-H1-shRNA vector. The anti-PRL antibody were obtained through the National Hormone and Peptide Program (NHPP), National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK), and Dr. Parlow (NHPP). Monoclonal antibody G3G4 BrdU was developed by Kaufman, S. J. and obtained from the Developmental Studies Hybridoma Bank developed under the auspices of the NICHD and maintained by the University of Iowa, Department of Biological Sciences, Iowa City, IA 52242.

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Copyright information

© Springer Science+Business Media B.V. 2009

Authors and Affiliations

  • Guoxin Wang
    • 1
  • Dawei Chen
    • 1
  • Haoshu Luo
    • 1
  • Jiali Liu
    • 1
  • Xiaowen Ji
    • 1
  • Jingjing Fan
    • 1
  • Sheng Cui
    • 1
  1. 1.State Key Laboratories for AgroBiotechnology, College of Biological SciencesChina Agricultural UniversityHaidian District BeijingPeople’s Republic of China

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