Melatonin-induced estrogen receptor alpha-mediated calbindin-D9k expression plays a role in H2O2-mediated cell death in rat pituitary GH3 cells.
Calbindin-D9k (CaBP-9k) is a 9-kDa polypeptide possessing two calcium-binding sites that is expressed in the mammalian intestine, uterus, and pituitary gland. The factors regulating the expression of the estrogen receptor (ER) and CaBP-9k in the pituitary gland are currently unknown. In this study, we investigated whether the ER and CaBP-9k expression are regulated by melatonin during H(2)O(2)-induced cell death in rat pituitary GH3 cells. Cell survival increased by approximately 27-36% in H(2)O(2) plus melatonin compared to H(2)O(2) alone, and CaBP-9k expression was augmented by treatment with H(2)O(2) plus melatonin. These results suggest that the increase in cell survival and the melatonin-induced CaBP-9k expression may play a role in protecting cells against H(2)O(2)-mediated cell death. This result is also consistent with the increase in CaBP-9k expression leading to rises in p-ERK and p-Bad (S112). Over-expression of CaBP-9k caused an increase in p-ERK. ERalpha expression was higher in H(2)O(2) plus melatonin-treated cells compared to those treated with H(2)O(2) alone, while ERbeta expression was not. Also, ERalpha in the nuclear fraction increased in the presence of melatonin and decreased in the presence of ICI 182 780 or ICI 182 780 plus melatonin. The relative binding affinity of ERalpha for melatonin was higher than that of ERbeta, suggesting that melatonin has the potential to preferentially bind ERalpha. In conclusion, these results indicate that melatonin may increase CaBP-9k expression through ERalpha.
Laboratory of Veterinary Biochemistry and Molecular Biology, College of Veterinary Medicine, Chungbuk National University, Chungbuk, Korea.
Yeong-Min
YM
Yoo
Eui-Bae
EB
Jeung
D000818:Animals; D015153:Blotting, Western; D064026:Calbindins; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D002492:Central Nervous System Depressants / Q000494:pharmacology*; D004958:Estradiol / Q000031:analogs & derivatives / Q000494:pharmacology; D047628:Estrogen Receptor alpha / Q000378:metabolism*; D000077267:Fulvestrant; D005786:Gene Expression Regulation / Q000187:drug effects*; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D047468:Immunoprecipitation; D008550:Melatonin / Q000494:pharmacology*; D016877:Oxidants / Q000494:pharmacology; D051381:Rats; D064030:S100 Calcium Binding Protein G / Q000378:metabolism*
D064026:Calbindins; D002492:Central Nervous System Depressants; D047628:Estrogen Receptor alpha; D016877:Oxidants; D064030:S100 Calcium Binding Protein G; C576508:S100g protein, rat; D000077267:Fulvestrant; D004958:Estradiol; D006861:Hydrogen Peroxide; D008550:Melatonin