Abstract
We have identified and quantified specific mRNAs in the human colonic carcinoma cell line Caco-2 by reverse transcriptase-polymerase chain reaction. Initial examination revealed that like rat duodenal mucosa, Caco-2 cells possessed mRNA for the vitamin D receptor. Using primers for human calbindin we found a 237-bp PCR product in Caco-2 cell RNA, but not from rat duodenal RNA. Primers for rat calbindin did not amplify calbindin mRNA in Caco-2 RNA, confirming a high degree of mismatch between rat and human sequences. 1 ,25(OH) 2 vitamin D 3 treatment (10 nM) significantly elevated calbindin mRNA levels 50% by 12 h, with maximal levels occurring by 48 h (fivefold elevation). Increasing concentrations of 1,25(OH) 2 vitamin D 3 (from 15 pM to 100 nM) caused progressive increases in calbindin mRNA levels following 48 h of treatment. Elevated calbindin mRNA levels were associated with enhancement of transcellular calcium transport. Our results are the first demonstration of vitamin D-regulated calbindin mRNA in a human intestinal cell line.
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