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CYP24 is a well-established vitamin D receptor (VDR) target gene. The active VDR ligand 1,25(OH) 2 D 3 regulates its own catabolism by increasing CYP24 expression. It is well known that in the presence of 1,25(OH) 2 D 3 , VDR binds to VDREs in the promoter region of CYP24 and initiates CYP24 transcription. However, little is known about the role of 1,25(OH) 2 D 3 in the posttranscriptional modulation of CYP24 . In this study, we investigated the functional significance of 1,25(OH) 2 D 3 in CYP24 RNA splicing in colon cancer cells. Using RT-PCR, we found that 1,25(OH) 2 D 3 actively induces CYP24 splicing in a time-dependent manner and CYP24 splicing pattern could be cell type or tissue specific. The induction of RNA splicing by 1,25(OH) 2 D 3 was mainly CYP24 selective. Treatment of cells with parathyroid hormone inhibited basal CYP24 splicing, but failed to inhibit 1,25(OH) 2 D 3 -induced CYP24 splicing. Further experiments demonstrated that new RNA synthesis was required for the induction of CYP24 splicing by vitamin D. In addition, alteration of multiple signaling pathways also affected CYP24 splicing and cellular sensitivity in response to vitamin D appeared to correlate with the induction of CYP24 splicing. These results suggest that 1,25(OH) 2 D 3 not only regulates C YP24 transcription, but also plays an important role in posttranscriptional modulation of CYP24 by inducing its splicing. Our findings reveal an additional regulatory step that makes the vitamin D mediated action more prompt and efficient.

Regulation of CYP24 splicing by 1,25-dihydroxyvitamin D3 in human colon cancer cells