Tomono Susumu, Sawada Natsumi, Yamaguchi Masayoshi
Laboratory of Endocrinology and Molecular Metabolism, Graduate School of Nutritional Sciences, University of Shizuoka, Shizuoka, Japan.
Int J Mol Med. 2007 Jul;20(1):79-83.
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.
一种新型蛋白质RGPR-p117被发现是一种与调钙素基因启动子区域相关的蛋白质,它能与TTGGC基序结合。已知调钙素可调节多种细胞类型中的细胞内信号系统。在克隆的正常大鼠肾近端小管上皮NRK52E细胞中,RGPR-p117已被证明可增强调钙素基因的启动子活性。然而,RGPR-p117在细胞功能中的作用仍有待阐明。本研究旨在确定RGPR-p117的过表达是否对NRK52E细胞的细胞增殖、蛋白质和DNA含量有影响。NRK52E细胞(野生型)或稳定转染RGPR-p117/phCMV2的细胞(转染子)在含有5%牛血清(BS)的杜尔贝科改良伊格尔培养基中培养。RGPR-p117在转染子中显著表达。NRK52E细胞(野生型)或转染子在含有5%BS的培养基中培养24、48或72小时,亚汇合后,细胞在不含BS的培养基中培养24、48或72小时。与野生型细胞相比,转染子中的细胞增殖没有显著变化。在有BS存在的情况下,随着细胞增殖,NRK52E细胞中的转染子中的蛋白质和DNA含量显著降低。当亚汇合的NRK52E细胞在不含BS的培养基中培养24、48或72小时时,与野生型细胞相比,转染子细胞的数量没有显著变化。亚汇合后在不含BS的培养基中培养的转染子中,NRK52E细胞中的蛋白质和DNA含量显著降低。本研究表明,RGPR-p117的过表达诱导NK52E细胞中蛋白质和DNA含量的降低,表明其在细胞功能调节中的作用。