Identification of a highly potent vitamin D receptor antagonist: (25S)-26-Adamantyl-25-hydroxy-2-methylene-22,23-didehydro-19,27-dinor-20-epi-vitamin D3 (ADMI3)

Vitamin D Receptor Activation Enhances Benzo[a]pyrene Metabolism via CYP1A1 Expression in Macrophages

“…1,25(OH) 2 D 3 , BaP, and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) were purchased from Wako Pure Chemical Industries (Osaka, Japan), and ␣-naphthoflavone was obtained from Sigma-Aldrich (St. Louis, MO). LCA acetate, (25R)- 25-adamantyl-1␣,25-dihydroxy-2-methylene-22,23-didehydro-19,26,27-trinor-20-epi-vitamin D 3 (ADTT) and (25S)- 26-adamantyl-1␣,25-dihydroxy-2-methylene-22,23-didehydro-19,27-dinor-20-epi-vitamin D 3 (ADMI3) were synthesized in the Sachiko Yamada laboratory (Igarashi et al, 2007;Ishizawa et al, 2008;Nakabayashi et al, 2008).…”

“…4B). In addition, we examined the effect of VDR antagonists, ADTT and ADMI3 (Igarashi et al, 2007;Nakabayashi et al, 2008). Both ADTT and ADMI3 effectively reduced CYP1A1 expression induced by BaP plus 1,25(OH) 2 D 3 in THP-1 cells (Fig.…”

Dynamic and Ligand-Selective Interactions of Vitamin D Receptor with Retinoid X Receptor and Cofactors in Living Cells

“…NM_006312) were inserted into pEYFP-C1 to make pEYFP-RXR, pEYFP-SRC-1 (ID1-ID3), pEYFP-SRC-1 (ID4), pEYFP-SMRT (ID1), pEYFP-SMRT (ID2), and pEYFP-SMRT (ID1 ϩ ID2), respectively. Fragments of SRC-1 (ID4), SMRT (ID1), SMRT (ID2), and SMRT (ID1 ϩ ID2) were inserted into the pCMX-GAL4 vector to make pCMX-GAL4-SRC-1 (ID4), pCMX-GAL4-SMRT (ID1), pCMX-GAL4-SMRT (ID2), and pCMX-GAL4-SMRT (ID1 ϩ ID2), respectively (Igarashi et al, 2007;Inaba et al, 2007). pCMX-VDR, pCMX-VP16-VDR, pCMX-GAL4-SRC-1 (ID1-ID3; amino acids 595-771), Sppx3-tk-LUC, and MH100(UAS)x4-tk-LUC were previously reported (Igarashi et al, 2007;Inaba et al, 2007).…”

“…For luciferase reporter assays, transfection in HEK293 cells was performed via calcium phosphate coprecipitation (Inaba et al, 2007). Transfection used 50 ng of a reporter plasmid (Sppx3-tk-LUC for VDR or MH100(UAS)x4-tk-LUC for GAL4), 15 ng of each expression plasmid, and 10 ng of pCMX-␤-galactosidase for each well of 96-well plate (Igarashi et al, 2007;Inaba et al, 2007). Luciferase data were normalized to an internal ␤-galactosidase control.…”

“…We transiently transfected HEK293 cells with an expression vector for AmCyan-VDR or EYFP-VDR (Fig. 1A) and a luciferase reporter containing a VDR-responsive direct repeat-3 element from the mouse osteopontin promoter (Igarashi et al, 2007) and measured 1,25(OH) 2 D 3 -dependent luciferase activity. 1,25(OH) 2 D 3 (100 nM) effectively induced transactivation of wild-type VDR, AmCyan-VDR, and EYFP-VDR but not of control empty vector, AmCyan, or EYFP (Fig.…”