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Am J Physiol Cell Physiol 280: C628-C636, 2001;
0363-6143/01 $5.00
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Vol. 280, Issue 3, C628-C636, March 2001

Regulation of the human sodium-phosphate cotransporter NaPi-IIb gene promoter by epidermal growth factor

Hua Xu, James F. Collins, Liqun Bai, Pawel R. Kiela, and Fayez K. Ghishan

Departments of Pediatrics and Physiology, Steele Memorial Children's Research Center, University of Arizona Health Sciences Center, Tucson, Arizona 85724

The intestinal sodium-phosphate cotransporter (NaPi-IIb) plays a major role in intestinal Pi absorption. Epidermal growth factor (EGF) is involved in the regulation of Pi homeostasis. However, the role of EGF in intestinal NaPi-IIb regulation is not clear. The current studies showed that EGF decreased NaPi-IIb mRNA abundance by 40-50% in both rat intestine and Caco-2 cells. To understand the mechanism of this regulation, we cloned the human NaPi-IIb gene and promoter region and studied the effect of EGF on NaPi-IIb gene transcription. The human NaPi-IIb gene has 12 exons and 11 introns. Two transcription initiation sites were identified by primer extension. Additionally, 2.8 kb of the 5'-flanking region of the gene was characterized as a functional promoter in human intestinal (Caco-2) and human lung (A549) cells. Additional studies showed that EGF inhibited promoter activity by 40-50% in Caco-2 cells and that actinomycin D treatment abolished this inhibition. EGF had no effect on promoter activity in lung (A549) cells. We conclude that the human NaPi-IIb gene promoter is functional in Caco-2 and A549 cells and that the gene is responsive to EGF by a transcriptionally mediated mechanism in intestinal cells.

Caco-2 cells; A549 cells; human intestine; sodium-phosphate cotransporter; transcriptional regulation


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