Hormonal regulation on regucalcin mRNA expression in cloned normal rat kidney proximal tubular epithelial NRK52E cells.
Regucalcin is a regulatory protein in cell signaling. This study was undertaken to determine whether regucalcin mRNA expresses in the cloned normal rat kidney proximal tubular epithelial NRK52E cells and its expression regulates due to hormones and cell signaling-related factors. Cells with subconfluency were cultured for 24, 48, or 72 h in a Dulbecco's modified Eagle medium supplemented with non-essential amino acid without bovine serum (BS). The result of Western blot analysis showed that regucalcin protein was present in the NRK52E cells. The expression of regucalcin mRNA in the cells was determined using reverse transcription-polymerase chain reaction (RT-PCR). Regucalcin mRNA expression in the NRK52E cells was significantly increased by culture with parathyroid hormone (PTH, 10(-8) or 10(-7) M), aldosterone (10(-8) or 10(-7) M), or dexamethasone (10(-8) M). The presence of 1,25-dihydroxyvitamin D(3) (1,25(OH)2D3, 10(-8) or 10(-7) M) or calcitonin (10(-9) or 10(-8) M) did not have a significant effect on regucalcin mRNA levels in the cells. Culture with dibutyryl cyclic AMP (DcAMP, 10(-5) or 10(-4) M) or phorbol 12-myristate 13-acetate (PMA, 10(-6) M), an activator of protein kinase C, caused a significant increase in regucalcin mRNA expression. The presence of staurosporine (10(-8) M) caused a significant decrease in regucalcin mRNA expression. Dibucaine (10(-7) M), PD98059 (10(-7) M), or vanadate (10(-6) or 10(-5) M) did not have an effect on regucalcin mRNA levels. The present study demonstrates that regucalcin mRNA and its protein are expressed in the cloned normal rat kidney proximal tubular epithelial NRK52E cells, and that the expression is enhanced by hormones which regulate ion transport in the proximal tubule.
Nakagawa
Taeko
T
Laboratory of Endocrinology and Molecular Metabolism, Graduate School of Nutritional Sciences, University of Shizuoka, 52-1 Yada, Shizuoka 422-8526, Japan.
Yamaguchi
Masayoshi
M
D000818:Animals; D002135:Calcium-Binding Proteins / Q000096:biosynthesis* / Q000235:genetics; D002265:Carboxylic Ester Hydrolases; D002460:Cell Line; D004847:Epithelial Cells / Q000166:cytology / Q000502:physiology*; D005786:Gene Expression Regulation / Q000187:drug effects* / Q000502:physiology; D006728:Hormones / Q000494:pharmacology*; D047908:Intracellular Signaling Peptides and Proteins; D007687:Kidney Tubules, Proximal / Q000166:cytology / Q000502:physiology*; D012333:RNA, Messenger / Q000096:biosynthesis* / Q000235:genetics; D051381:Rats; D015238:Sulfotransferases
D002135:Calcium-Binding Proteins; D006728:Hormones; D047908:Intracellular Signaling Peptides and Proteins; D012333:RNA, Messenger; D015238:Sulfotransferases; C028152:alcohol sulfotransferase; D002265:Carboxylic Ester Hydrolases; C513325:Rgn protein, rat