Effect of 17 beta-estradiol on the human osteosarcoma cell line MG-63.
We present evidence that 17 beta-estradiol (17 beta-E2) regulates 1,25(OH)2D3-induced alkaline phosphatase synthesis and osteocalcin secretion by the human osteosarcoma cell line MG-63. When cells were pre-treated with 17 beta-E2 for 48 h prior to treatment with 1,25(OH)2D3 (50 nM) for another 48 h, alkaline phosphatase activity increased by 40% (P < 0.025) with 2 nM 17 beta-E2 and plateaued at levels of 20 and 200 nM 17 beta-E2. Under the same experimental conditions, osteocalcin secretion was enhanced by 37% (P < 0.005) with 2 nM E2. However, 17 beta-E2 had no effect on basal alkaline phosphatase or on osteocalcin secretion. Moreover, simultaneous addition of 17 beta-E2 and 1,25(OH)2D3 to cells did not result in any additional effect over 1,25(OH)2D3 treatment alone. Tamoxifen (10 nM) inhibited 17 beta-E2-induced activities in 1,25(OH)2D3-treated cells while not affecting control cells. Dexamethasone pretreatment (100 nM, 48 h) also stimulated alkaline phosphatase activity in MG-63 cells. Moreover, dexamethasone pretreatment followed by treatment with 17 beta-E2 and 1,25(OH)2D3 gave an additive effect for alkaline phosphatase activity. 17 alpha-Estradiol (17 alpha-E2), a less active form of estrogen, failed to modify, at low concentrations, control or 1,25(OH)2D3-induced alkaline phosphatase synthesis and osteocalcin secretion. In fact, a 100-1000-fold higher concentration of 17 alpha-E2 was necessary to reproduce the effects of 17 beta-E2 on osteocalcin secretion. The addition of insulin-like growth factor I (IGF-I) for 24 h (1-50 ng/ml) to MG-63 cells did not modify 1,25(OH)2D3-induced osteocalcin release from these cells. However, longer incubations with 50 ng/ml IGF-I did reproduce some of the effects observed with 17 beta-E2. Thus, the effects of 17 beta-E2 are probably not related to IGF-I production in MG-63 cells since under these conditions the addition of IGF-I alone should have produced a response at shorter incubation times and in the presence of lower concentrations of IGF-I. Since 17 beta-E2 pretreatment was necessary to observe any effects on 1,25(OH)2D3-induced activities, we hypothesized that 17 beta-E2 regulated 1,25(OH)2D3 receptors in MG-63 cells. When cells were treated with 100 nM 17 beta-E2 for 48 h, the binding affinity was unchanged: 37.3 +/- 1.9 versus 35.1 +/- 0.4 pM for cells whether treated or not with 17 beta-E2, respectively. In contrast, a significant increase in binding capacity (Bmax) was noted (15 +/- 3.5%; P < 0.025).(ABSTRACT TRUNCATED AT 400 WORDS)
Lajeunesse
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Hôpital Maisonneuve-Rosemont, Centre de Recherche, Montréal, Canada.
D000469:Alkaline Phosphatase / Q000096:biosynthesis / Q000187:drug effects; D000704:Analysis of Variance; D002455:Cell Division / Q000187:drug effects; D004347:Drug Interactions; D004958:Estradiol / Q000494:pharmacology*; D006801:Humans; D007334:Insulin-Like Growth Factor I / Q000494:pharmacology; D010006:Osteoblasts / Q000187:drug effects* / Q000378:metabolism; D015675:Osteocalcin / Q000378:metabolism; D012516:Osteosarcoma; D018167:Receptors, Calcitriol / Q000187:drug effects; D014407:Tumor Cells, Cultured
D018167:Receptors, Calcitriol; D015675:Osteocalcin; D004958:Estradiol; D007334:Insulin-Like Growth Factor I; D000469:Alkaline Phosphatase