Overexpression of regucalcin suppresses cell proliferation of cloned normal rat kidney proximal tubular epithelial NRK52E cells

  • Authors:
    • Taeko Nakagawa
    • Natumi Sawada
    • Masayoshi Yamaguchi
  • View Affiliations

  • Published online on: October 1, 2005     https://doi.org/10.3892/ijmm.16.4.637
  • Pages: 637-643
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Abstract

The role of regucalcin, which is a regulatory protein in intracellular signaling pathway, in the regulation of cell proliferation was investigated by using the cloned normal rat kidney proximal tubular epithelial NRK52E cells over-expressing regucalcin. NRK52E cells were transfected with regucalcin (RC) pCXN2 vector and the multiple neomycin-resistant clones that stably overexpress regucalcin were selected. The regucalcin content of RC/pCXN2-transfected cells used in this study was about 21-fold as compared with that of the parental wild-type NRK52E cells. Wild-type NRK52E cells, pCXN2 vector-transfected cells (mock-type), and RC/ pCXN2-transfected cells (transfectants) were cultured for 24, 48, and 72 h in the presence of bovine serum (5%). The cell numbers of wild- and mock-type were significantly increased with the time course of the culture. Cell numbers of transfectants were significantly suppressed as compared with that of wild- and mock-type. The decrease in cell number of wild-type cultured for 72 h in the presence of butyrate (8.3x10-6 or 8.3x10-5 M), rescovitine (10-8 or 10-7 M), or sulforaphane (10-9 M), which is an inhibitor of the cell cycle, was not observed in transfectants. The effect of PD98059 (10-8 M), staurosporine (10-10 M) or dibucaine (10-8-10-6 M), which is an inhibitor of protein kinases, in decreasing cell number of wild-type was not seen in transfectants. Moreover, the culture with wortmannin (10-8 or 10-7 M), an inhibitor of phosphatidylinositol 3 (PI3)-kinase, or Bay K 8644 (10-8 or 10-7 M), an agonist of calcium entry in cells, caused a significant decrease in cell number of the wild-type. This decrease was not observed in transfectants. The result of reverse transcription-polymerase chain reaction (RT-PCR) analysis using specific primers showed that c-jun and chk2 mRNA levels were significantly decreased in transfectant. p53 mRNA level was significantly increased in transfectants. The expression of c-myc, c-fos, cdc2, p21, and G3PDH mRNAs in transfectants was not significantly changed. This study demonstrates that overexpression of regucalcin has a suppressive effect on cell proliferation, which is mediated through various signaling pathways, in the cloned normal rat kidney proximal tubular epithelial NRK52E cells.

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October 2005
Volume 16 Issue 4

Print ISSN: 1107-3756
Online ISSN:1791-244X

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Spandidos Publications style
Nakagawa T, Sawada N and Yamaguchi M: Overexpression of regucalcin suppresses cell proliferation of cloned normal rat kidney proximal tubular epithelial NRK52E cells. Int J Mol Med 16: 637-643, 2005
APA
Nakagawa, T., Sawada, N., & Yamaguchi, M. (2005). Overexpression of regucalcin suppresses cell proliferation of cloned normal rat kidney proximal tubular epithelial NRK52E cells. International Journal of Molecular Medicine, 16, 637-643. https://doi.org/10.3892/ijmm.16.4.637
MLA
Nakagawa, T., Sawada, N., Yamaguchi, M."Overexpression of regucalcin suppresses cell proliferation of cloned normal rat kidney proximal tubular epithelial NRK52E cells". International Journal of Molecular Medicine 16.4 (2005): 637-643.
Chicago
Nakagawa, T., Sawada, N., Yamaguchi, M."Overexpression of regucalcin suppresses cell proliferation of cloned normal rat kidney proximal tubular epithelial NRK52E cells". International Journal of Molecular Medicine 16, no. 4 (2005): 637-643. https://doi.org/10.3892/ijmm.16.4.637