Overexpression of RGPR-p117 induces the decrease in protein and DNA contents in cloned normal rat kidney proximal tubular epithelial NRK52E cells.
A novel protein RGPR-p117 was discovered as a regucalcin gene promoter region-related protein that binds to the TTGGC motif. Regucalcin is known to regulate the intracellular signaling system in many cell types. RGPR-p117 has been shown to enhance the promoter activity of the regucalcin gene in cloned normal rat kidney proximal tubular epithelial NRK52E cells. The role of RGPR-p117 in cell function remains to be elucidated, however. This study was undertaken to determine whether overexpression of RGPR-p117 has an effect on cell proliferation, protein and DNA contents in NRK52E cells. NRK52E cells (wild-type) or stable RGPR-p117/phCMV2-transfected cells (transfectants) were cultured in Dulbecco's minimum essential medium containing 5% bovine serum (BS). RGPR-p117 was markedly expressed in the transfectants. NRK52E cells (wild-type) or transfectants were cultured for 24, 48, or 72 h in a medium containing 5% BS, and after subconfluency the cells were cultured for 24, 48, or 72 h in a medium without BS. Cell proliferation was not significantly changed in the transfectants as compared with that of wild-type cells. Protein and DNA contents in NRK52E cells were significantly decreased in the transfectants with cell proliferation in the presence of BS. When NRK52E cells with subconfluency were cultured for 24, 48, or 72 h in a medium without BS, the number of transfectant cells was not significantly changed compared with that of wild-type cells. Protein and DNA contents in NRK52E cells were significantly decreased in the transfectants cultured in a medium without BS after subconfluency. This study demonstrates that overexpression of RGPR-p117 induces the decrease in protein and DNA contents in NK52E cells, indicating its role in the regulation of cell function.
Tomono
Susumu
S
Laboratory of Endocrinology and Molecular Metabolism, Graduate School of Nutritional Sciences, University of Shizuoka, Shizuoka, Japan.
Sawada
Natsumi
N
Yamaguchi
Masayoshi
M
D000818:Animals; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D002999:Clone Cells; D016895:Culture Media, Serum-Free; D004247:DNA / Q000032:analysis*; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000494:pharmacology; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects; D007687:Kidney Tubules, Proximal / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D011506:Proteins / Q000032:analysis*; D051381:Rats; D013997:Time Factors; D014162:Transfection
D016895:Culture Media, Serum-Free; D004268:DNA-Binding Proteins; D011506:Proteins; C486711:Sec16b protein, rat; D004247:DNA