We compared the effects of 1α,25-dihydroxyvitamin D
3 [1α,25(OH)
2D
3] and its analog, 1α,25-dihydroxy-16-ene-vitamin D
3 [1α,25(OH)
2-16-ene-D
3], as well as their interactions with 17-β estradiol (E
2) on osteoblastic function in our human normal (HOB) and osteosarcoma SaOS-2 cell models representing two different stages of differentiation, the more differentiated HOB+DEX cells and SaOS+DEX cells, and the corresponding less differentiated HOB-DEX and SaOS-DEX cells. The differential effects of 1α,25(OH)
2D
3 and 1α,25(OH)
2-16-ene-D
3 and the modulation by E
2 on ALP activity in HOB-DEX and HOB+DEX cells were small but significant. The most significant effects were seen in SaOS+DEX cells, in which 1α,25(OH)
2-16-ene-D
3 was 100-fold more potent than 1α,25(OH)
2D
3, the maximal enhancement being exerted at 0.1 nM and 10 nM, respectively. E
2 enhanced the stimulatory effects of both compounds, with ALP being increased 2-fold at 0.1 nM (p<0.001). Osteocalcin (OC) production in HOB-DEX cells was stimulated 1.3 to 1.4-fold by 1α,25(OH)
2D
3 and 1α,25(OH)
2-16-ene-D
3 at a concentration of 0.01 nM, with E
2 inhibiting the effect of 1α,25(OH)
2-16-ene-D
3. In SaOS-DEX and SaOS+DEX cells, 1α,25(OH)
2D
3 and 1α,25(OH)
2-16-ene-D
3 stimulated OC production 1.6-fold at 0.1 nM with E
2 slightly enhancing the effect of 1α,25(OH)
2D
3. Western blot analysis of 1α,25(OH)
2D
3 receptor (VDR) levels showed that in SaOS+DEX cells, the effect of 1α,25(OH)
2D
3 was larger than that of 1α,25(OH)
2-16-ene-D
3. These results show that 1α,25(OH)
2-16-ene-D
3 is biologically active in human osteoblasts.
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