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Am J Physiol Renal Physiol (December 3, 2002). doi:10.1152/ajprenal.00252.2002
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Articles in PresS, published online ahead of print December 3, 2002
Am J Physiol Renal Physiol, 10.1152/ajprenal.00252.2002
Submitted on July 12, 2002
Accepted on November 29, 2002

HCaRG is a novel regulator of renal epithelial cell growth and differentiation causing G2M arrest

Alison M. Devlin1, Nicolas Solban1, Sandra Tremblay1, Jolanta Gutkowska1, Walter Schurch1, Sergei N. Orlov1, Richard Lewanczuk2, Pavel Hamet1, and Johanne Tremblay1*

1 Laboratory of Cellular Biology of Hypertension, Centre de Recherche, Centre Hospitalier de l'Universite de Montreal (CHUM), Montreal, Quebec, Canada
2 Department of Endocrinology, University of Alberta, Edmonton, Alberta, Canada

* To whom correspondence should be addressed. E-mail: johanne.tremblay{at}umontreal.ca.

We recently identified a novel calcium-regulated gene; HCaRG, which is highly expressed in the kidney and maps to a chromosomal locus determining kidney weight in the rat. The mRNA levels of HCaRG negatively correlate with the proliferative status of the kidney cells. In order to investigate its role in renal epithelial cellular growth directly, we have studied the human embryonic kidney cell line (HEK 293) stably transfected with either plasmid alone or plasmid containing rat HCaRG. [3H]-Thymidine incorporation was significantly lower in HCaRG clones. Although HCaRG clones exhibited some enhanced susceptibility to cell death, this was not the primary mechanism of reduced proliferation. Cell cycle analysis revealed a G2M phase accumulation in HCaRG clones which was associated with upregulation of p21Cip1/WAF-1 and down-regulation of p27Kip1. HCaRG clones had a greater protein content, larger cell size and released 4.5-8 fold more of an ANP-like immunoreactivity compared to controls. In addition, HCaRG clones demonstrated the presence of differentiated junctions and a lower incidence of mitotic figures. Genistein treatment of wild-type HEK 293 cells mimicked several phenotypic characteristics associated with HCaRG overexpresssion including increased cell size and increased release of ANP. Taken together, our results suggest that HCaRG is a regulator of renal epithelial cell growth and differentiation causing G2M cell cycle arrest.




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