Phytoestrogens regulate transcription and translation of vitamin D receptor in colon cancer cells.
The present study assesses the effects of two isoflavones, genistein and glycitein, and equol - a product of intestinal bacterial metabolism of dietary isoflavones, on vitamin D receptor (VDR) expression in an intestinal HT29 cell line. Genistein and glycitein significantly upregulated the VDR transcription and translation in HT29 cells. The effect of equol was less pronounced. Treating HT29 cells transfected with a vector containing the VDR promoter next to a luciferase reporter with genistein or glycitein resulted in significant upregulation of VDR promoter activity, in a manner similar to that induced by 17beta-estradiol (E2). Again, the effect of equol was less pronounced. VDR luciferase promoter activity was upregulated most by genistein, then by glycitein and least by equol when the VDR promoter was cotransfected with estrogen receptor beta. Reporter gene and chromatin immunoprecipitation (ChIP) assays demonstrated that E2 upregulates AP-1 and Sp-1 sites present on the VDR gene. In contrast, the same assays demonstrated that the Sp-1, but not AP-1, site is induced by the phytoestrogens. Similar to E2, genistein, glycitein and the isoflavonoid metabolite equol induced higher concentrations of intracellular free calcium, an event that could provide the upstream mechanism(s) induced by E2 and phytoestrogens that initiates the signaling cascade which results in the activation of extracellular signal-regulated kinase (ERK) signaling pathways and modulation of Sp-1 sites of the VDR gene, and culminates in enhanced VDR expression.
Gilad
Liat Abovich
LA
Faculty of Agricultural, Food and Environmental Quality Sciences, Institute of Biochemistry, Food Science and Nutrition, The Hebrew University of Jerusalem, Rehovot 76100, Israel.
Tirosh
Oren
O
Schwartz
Betty
B
D000230:Adenocarcinoma / Q000378:metabolism*; D015153:Blotting, Western / Q000379:methods; D002118:Calcium / Q000378:metabolism; D003110:Colonic Neoplasms / Q000378:metabolism*; D060754:Equol; D004958:Estradiol / Q000494:pharmacology; D047629:Estrogen Receptor beta / Q000235:genetics / Q000378:metabolism; D019833:Genistein / Q000494:pharmacology; D019073:HT29 Cells; D006801:Humans; D047468:Immunoprecipitation; D007529:Isoflavones / Q000494:pharmacology; D048789:Phytoestrogens / Q000494:pharmacology*; D011401:Promoter Regions, Genetic; D014176:Protein Biosynthesis; D018167:Receptors, Calcitriol / Q000235:genetics* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016329:Sp1 Transcription Factor / Q000235:genetics / Q000378:metabolism; D018808:Transcription Factor AP-1 / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic; D014162:Transfection / Q000379:methods
C007610:4',7-dihydroxy-3,4-dihydroisoflavone; D047629:Estrogen Receptor beta; D007529:Isoflavones; D048789:Phytoestrogens; D018167:Receptors, Calcitriol; D016329:Sp1 Transcription Factor; D018808:Transcription Factor AP-1; D004958:Estradiol; D060754:Equol; C086566:glycitein; D019833:Genistein; D002118:Calcium