Effect of 1,25(OH)2D3 and its analogues on membrane phosphoinositide turnover and [Ca2+]i in Caco-2 cells.
Our laboratory has recently reported that 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] rapidly stimulates membrane polyphosphoinositide turnover and increases intracellular calcium concentration ([Ca2+]i) in Caco-2 cells. The role of binding to the vitamin D receptor (VDR) in the regulation of these rapid biochemical events, however, remains unclear. The present studies were, therefore, conducted using analogues of 1,25(OH)2D3, which differ markedly in their affinities for the VDR, to assess and compare their effects on [Ca2+]i and on inositol 1,4,5-trisphosphate (IP3) formation. Competitive binding studies performed with both intact cells and high-salt cytosolic extracts from Caco-2 cells demonstrated that 1,25(OH)2D3 and 1,24-(OH)2-22-ene-24-cyclopropyl-D3 (BT) have high affinities for the VDR; 25(OH)-16-ene,23-yne-D3 (AT), however, has a much lower affinity (approximately 1,000-fold less) for the VDR. Despite these large differences in binding affinities for the VDR, AT and BT produced similar concentration-dependent increases in [Ca2+]i and in IP3 formation while 1,25(OH)2D3 was approximately 10-fold less active. These results indicate that the structural requirements for the rapid action of these secosteroids on signal transduction in Caco-2 cells are different from those for receptor binding and transcriptional regulation.
Tien
X Y
XY
Department of Medicine, University of Chicago, Illinois 60637.
Brasitus
T A
TA
Qasawa
B M
BM
Norman
A W
AW
Sitrin
M D
MD
D001667:Binding, Competitive; D002117:Calcitriol / Q000031:analogs & derivatives / Q000378:metabolism / Q000494:pharmacology*; D002118:Calcium / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism; D006801:Humans; D015544:Inositol 1,4,5-Trisphosphate / Q000096:biosynthesis; D007425:Intracellular Membranes / Q000378:metabolism*; D009994:Osmolar Concentration; D010716:Phosphatidylinositols / Q000378:metabolism*; D018167:Receptors, Calcitriol; D011987:Receptors, Steroid / Q000378:metabolism; D014407:Tumor Cells, Cultured
D010716:Phosphatidylinositols; D018167:Receptors, Calcitriol; D011987:Receptors, Steroid; D015544:Inositol 1,4,5-Trisphosphate; D002117:Calcitriol; D002118:Calcium