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Articles in PresS, published online ahead of print December 19, 2001
Am J Physiol Gastrointest Liver Physiol, 10.1152/ajpgi.00269.2001
Submitted on June 21, 2001
Accepted on December 18, 2001
1 Department of Foods and Nutrition, Purdue University, West Lafayette, IN, USA
2 Graduate Program in Nutrition, University of North Carolina at Greensboro, Greensboro, NC, USA
3 Mineral Bioavailability Laboratory, Tufts University, Boston, MA, USA
* To whom correspondence should be addressed. E-mail: fleetj{at}cfs.purdue.edu.
The parental cell line (P) of Caco-2 cells and two clones, BBe and TC7, were studied at 11 days post-confluence to test the facilitated diffusion model of vitamin D-mediated, intestinal calcium absorption (CaTx). Nuclear vitamin D receptor (nVDR) and calbindin D9k (CaBP) were measured by Western blot, 24-hydroxylase (CYP24), CaBP, Ca ATPase (PMCA), and Ca channel (CaT1) mRNA levels were examined by RT-PCR, and net apical-to-basolateral CaTx was examined after treating cells for +/- 10 nM calcitriol for 8 hours (mRNA levels) or 48 hours (protein, CaBP mRNA, CaTx). nVDR level was lowest in BBe (38% P value) and directly related to CYP24 induction (TC7 = P 1.56X > BBe). nVDR was inversely related to the vitamin D-induced levels of CaT1 mRNA, CaBP mRNA, PMCA mRNA, and net CaTx with the highest induction seen in BBe. Basal CaBP mRNA (86X>P) and protein levels were highest in TC7 cells and were not associated with higher net CaTx suggesting CaBP may not be rate limiting for CaTx in these cells.
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