Difficulty Demonstrating Estradiol-Mediated Erk1/2 Phosphorylation in MCF-7 Cells

Details

• Personal Author:
  Antonini JM ; Brower SL ; Miller MR ; Roberts, Jennifer R.
• Description:
  While some studies report that estradiol (E2) activates extracellular-signal regulated kinase (Erk1/2) in MCF-7 breast cancer cells, others report E2 does not activate this signaling pathway. This study attempted to resolve the conflicting reports by investigating experimental variables that could impact Erk1/2 activation using a high through-put assay that quantitatively assessed Erk1/2 phosphorylation. Variables tested included: cell staging and dosing regimes with and without charcoal-stripped serum, different MCF-7 cell sublines and culture densities and several E2 formulations and solvents. Levels of phosphorylated Erk1/2 were normalized to cellular protein rather than to total Erk1/2 protein because an antibody purported to recognize total Erk1/2 preferentially reacted with non-phosphorylated Erk1/2, potentially exaggerating the apparent level of Erk1/2 activation. Dosing MCF-7 cells with E2 containing small amounts of stripped serum induced Erk1/2 phosphorylation; however, this induction was largely attributed to serum factors. E2 administered in serum-free medium did not significantly alter Erk1/2 phosphorylation under any condition tested; immunocytochemistry corroborated this conclusion. While phosphatase inhibitors generally increased Erk1/2 phosphorylation, they did not impact E2-altered Erk1/2 phosphorylation. It remains important to resolve the basis of conflicting reports regarding E2-induced Erk1/2 activation due to the potential importance of this pathway on breast cancer and other processes. [Description provided by NIOSH]
• Subjects:
  Cells, Cultured Neoplasms Occupational Health Safety Solvents
• Keywords:
  Breast-cancer Cancer Cell-culture-techniques Cell-cultures Quantitative-analysis
• ISSN:
  0960-0760
• Document Type:
  Text
• Genre:
  Journal Article
• Place as Subject:
  OSHA Region 3 ; West Virginia
• CIO:
  National Institute for Occupational Safety and Health (NIOSH)
• Division:
  HELD - Health Effects Laboratory Division
• Topic:
  Workplace Safety & Health
• Location:
  North America
• Pages in Document:
  375-385
• Volume:
  96
• Issue:
  5
• NIOSHTIC Number:
  nn:20028684
• Citation:
  J Steroid Biochem Mol Biol 2005 Sep; 96(5):375-385
• Contact Point Address:
  Department of Biochemistry and Molecular Pharmacology, Mary Babb Randolph Cancer Center, P.O. Box 9142, West Virginia University Health Sciences Center, Morgantown, WV 26506-9142, USA
• Email:
  mmiller@hsc.wvu.edu
• Federal Fiscal Year:
  2005
• NORA Priority Area:
  Work Environment and Workforce: Mixed Exposures
• Peer Reviewed:
  True
• Source Full Name:
  The Journal of Steroid Biochemistry and Molecular Biology
• Collection(s):
  National Institute for Occupational Safety and Health
• Main Document Checksum:
  urn:sha-512:1dca917a9e4f920f8433e58987b9e4d2904675201a60da20789782b89ddf105d5fafa6366b903b1b6b7b928cf0dabc7a0a4cf0dcf63194176fcd589655dfde5e
• Download URL:
  https://stacks.cdc.gov/view/cdc/189883/cdc_189883_DS1.pdf
• File Type:
  [PDF - 656.75 KB ]