Estrogen receptor transactivation in MCF-7 breast cancer cells by melatonin and growth factors.
The pineal hormone, melatonin, inhibits proliferation of estrogen receptor (ER)-positive MCF-7 human breast cancer cells, modulates both ER mRNA and protein expression, and appears to be serum dependent, indicating interaction between melatonin and serum components. To examine the effects of melatonin on ER activity, ER transactivation assays were performed by transiently transfecting MCF-7 cells with an ERE-luciferase reporter construct. MCF-7 cells pre-treated with melatonin for as little as 5 min followed by either epidermal growth factor (EGF) or insulin resulted in the estrogen-independent transactivation of the ER. None of the compounds when used alone transactivated the ER. The ability of melatonin and EGF to transactivate the ER was abolished by the addition of the antiestrogen, ICI 164384, suggesting that melatonin and EGF co-operate to transactivate the ER. The modulation of ER transactivation was associated with changes in mitogen activated protein kinase activity and ER phosphorylation. This ER transactivation was blocked by pertussis toxin, a Galpha i-protein-coupled receptor inhibitor, suggesting cross talk between the G-protein-coupled melatonin receptor pathway and the EGF/insulin tyrosine kinase receptor pathways in modulating ER transactivation. Exactly how the ability of melatonin in combination with EGF to transactivate the ER relates to melatonin's observed growth suppressive effects is not clear. It is possible that, although melatonin and EGF transactivate the ER, this transactivation does not result in the full transcription of estrogen-responsive genes, but rather, makes the ER refractory to activation by estradiol, thus, blocking the mitogenic actions of estradiol.
Ram
P T
PT
Department of Anatomy, Tulane University School of Medicine, New Orleans, LA 70112, USA.
Kiefer
T
T
Silverman
M
M
Song
Y
Y
Brown
G M
GM
Hill
S M
SM
D015153:Blotting, Western; D001943:Breast Neoplasms; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000378:metabolism; D004815:Epidermal Growth Factor / Q000494:pharmacology; D004958:Estradiol / Q000031:analogs & derivatives / Q000494:pharmacology; D004965:Estrogen Antagonists / Q000494:pharmacology; D005260:Female; D006133:Growth Substances / Q000494:pharmacology*; D006801:Humans; D007328:Insulin / Q000494:pharmacology; D008550:Melatonin / Q000494:pharmacology*; D037342:Pertussis Toxin; D010766:Phosphorylation / Q000187:drug effects; D053284:Polyunsaturated Alkamides; D011956:Receptors, Cell Surface / Q000235:genetics; D018160:Receptors, Cytoplasmic and Nuclear / Q000235:genetics; D011960:Receptors, Estrogen / Q000235:genetics* / Q000378:metabolism; D044094:Receptors, Melatonin; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015533:Transcriptional Activation / Q000235:genetics*; D014162:Transfection; D014407:Tumor Cells, Cultured; D010566:Virulence Factors, Bordetella / Q000494:pharmacology
D004965:Estrogen Antagonists; D006133:Growth Substances; D007328:Insulin; D053284:Polyunsaturated Alkamides; D011956:Receptors, Cell Surface; D018160:Receptors, Cytoplasmic and Nuclear; D011960:Receptors, Estrogen; D044094:Receptors, Melatonin; D010566:Virulence Factors, Bordetella; D004958:Estradiol; D004815:Epidermal Growth Factor; C051781:ICI 164384; D037342:Pertussis Toxin; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D008550:Melatonin