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Journal of Chinese Integrative Medicine ›› 2011, Vol. 9 ›› Issue (11): 1248-1253.doi: 10.3736/jcim20111114

• Original Experimental Research • Previous Articles     Next Articles

Effects of daidzein on steroid receptor coactivator-1 expression in MC3T3-E1 cells and the mechanism

Li-fang Wang1, Jin-feng Wang1, Wei-fang Jin1, Hong-fu Wang1, Shao-fen Zhang2, Jian-jun Gao1()   

  1. 1. Department of Bone Metabolism, Institute of Radiation Medicine, Fudan University, Shanghai 200032, China
    2. Department of Gynecology, Obstetrics and Gynecology Hospital, Fudan University, Shanghai 200011, China
  • Received:2011-06-22 Accepted:2011-07-15 Online:2011-11-20 Published:2011-11-15

Objective: To investigate the roles of daidzein in the expressions of steroid receptor coactivator-1 (SRC-1) and nuclear receptor corepressor (NcoR) in MC3T3-E1 osteoblastic cells.
Methods: MC3T3-E1 cells were cultured in α-minimal essential medium (α-MEM) containing 2% fetal bovine serum and treated with various concentrations of daidzein (10-9, 10-7and 10-5mol/L) or 17β-estradiol at 10-8 mol/L for 3 d. The protein levels of SRC-1 and NcoR in MC3T3-E1 cells were determined by Western blotting. Estrogen receptor (ER) antagonist ICI182780 at 10-7 mol/L or specific ERα antagonist methyl-piperidino-pyrazole (MPP) at 10-6 mol/L were used to block the corresponding receptors, and then MC3T3-E1 cells were treated with daidzein at 10-7 mol/L or 10-5 mol/L for 3 d. SRC-1 and NcoR protein levels were detected by Western blotting.
Results: The protein levels of SRC-1 increased by 2.5 fold (P<0.05) and 2 fold (P<0.05) by 10-7 and 10-5 mol/L of daidzein respectively, while the NcoR levels were not significantly altered. 17β-Estradiol at dose of 10-8mol/L did not affect the expression of SRC-1 but decreased NcoR protein expression by 35% (P<0.05). Compared with the control, daidzein at 10-7and 10-5 mol/L did not increase SRC-1 expression when ERs were blocked by antagonist ICI182780. Daidzein at 10-7 and 10-5 mol/L up-regulated SRC-1 by 1.8 fold (P<0.05) and 2.4 fold (P<0.05) respectively while ERα was blocked by MPP.
Conclusion: Daidzein increases protein level of SRC-1 and the ratio of SRC-1/NcoR. ERβ, instead of ERα, participates in the action of daidzein in regulating SRC-1 expression. Up-regulation of SRC-1 and increase of SRC-1/NcoR are part of the mechanism of the estrogenic effect of daidzein in improving osteogenesis.

Key words: daidzein, steroid receptor coactivator-1, nuclear receptor corepressor, osteoblasts, receptors, estrogen

Figure 1

Expressions of SRC-1 and NcoR in MC3T3-E1 cells The specific bands and the relative protein levels of SRC-1 (A) and NcoR (B) in MC3T3-E1 cells after 3 days of treatment with 10-9 mol/L (D9), 10-7 mol/L (D7) and 10-5 mol/L (D5) of daidzein or 10-8 mol/L of 17β-estroldal (E8) respectively as determined by Western blotting. Data are presented relative to the control (Ctrl) gene expression level after normalization to GAPDH and expressed as mean±standard deviation (n=4; *P<0.05, vs Ctrl group). SRC-1: steroid receptor coactivator-1; NcoR: nuclear receptor corepressor; GAPDH: glyceraldehyde-3-phosphate dehydrogenase."

Figure 2

Roles of ERs in up-regulation of SRC-1 expression induced by daidzein The relative protein levels of SRC-1 (A) and NcoR (B) and the changes of ratio of SRC-1/NcoR (C) in MC3T3-E1 cells after 3 days of treatment with 10-7 (D7) or 10-5 mol/L (D5) of daidzein in combination with ICI182780 (10-7 mol/L) or MPP (10-6 mol/L) as determined by Western blotting. Data are presented relative to the control (Ctrl) gene expression level after normalization to GAPDH and expressed as mean±standard deviation (n=4; *P<0.05, vs Ctrl group). SRC-1: steroid receptor coactivator-1; ERs: estrogen receptors; NcoR: nuclear receptor corepressor; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; MPP: methyl-piperidino-pyrazole; ICI: ICI182780 (Faslodex)."

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