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Expression of calcium-binding protein regucalcin mRNA in the cloned human hepatoma cells (HegG2): Stimulation by insulin

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Abstract

The expression of hepatic Ca2+-binding protein regucalcin in the cloned human hepatoma cells (HepG2) was investigated. The change in regucalcin mRNA levels was analyzed by Northern blotting using rat liver regucalcin complementary DNA (0.9 kb of open reading frame). Regucalcin mRNA was expressed in HepG2 cells, although the mRNA was markedly expressed in normal rat liver. Moreover, regucalcin protein in HepG2 cells was detected by Western blot analysis using a polyclonal rabbit anti-regucalcin antibody. Regucalcin mRNA expression in HepG2 cells was clearly stimulated by the culture with insulin (10-8 M) of the effective concentration. Regucalcin protein in HepG2 cells was also increased by the treatment of insulin (10-8 M). The present results demonstrate that regucalcin is expressed in the transformed HepG2 cells, and that the expression is stimulated by insulin.

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References

  1. Yamaguchi M, Yamamoto T: Purification of calcium binding substance from soluble fraction of normal rat liver. Chem Pharm Bull 21: 1915–1918, 1978

    Google Scholar 

  2. Yamaguchi M: Physicochemical properties of calcium-binding protein isolated from rat liver cytosol: Ca2+-induced conformational changes. Chem Pharm Bull 36: 286–290, 1988

    Google Scholar 

  3. Yamaguchi M: A novel Ca2+-binding protein regucalcin and calcium inhibition: Regulatory role in liver cell function. In: K Kohama (ed). Calcium Inhibition. Japan Sci Soc Press, Boca Raton, 1992, pp 19–41

    Google Scholar 

  4. Yamaguchi M, Tai H: Inhibitory effect of calcium-binding protein regucalcin on Ca2+/calmodulin-dependent cyclic nucleotide phosphodiesterase activity in rat liver cytosol. Mol Cell Biochem 106: 25–30, 1991

    Google Scholar 

  5. Yamaguchi M, Mori S: Inhibitory effect of calcium-binding protein regucalcin on protein kinase C activity in rat liver cytosol. Biochem Med Metab Biol 43: 140–146, 1990

    Google Scholar 

  6. Yamaguchi M, Sakurai T: Inhibitory effect of calcium-binding protein regucalcin on Ca2+-activated DNA fragmentation in rat liver nuclei. FEBS Lett 279: 281–284, 1991

    Google Scholar 

  7. Shimokawa N, Yamaguchi M: Molecular cloning and sequencing of the cDNA coding for a calcium-binding protein regucalcin from rat liver. FEBS Lett 327: 251–255, 1993

    Google Scholar 

  8. Shimokawa N, Matsuda Y, Yamaguchi M: Genomic cloning and chromosomal assignment of rat regucalcin gene. Mol Cell Biochem 151: 157–163, 1995

    Google Scholar 

  9. Shimokawa N, Isogai M, Yamaguchi M: Specific species and tissue differences for the gene expression of calcium-binding protein regucalcin. Mol Cell Biochem 143: 67–71, 1995

    Google Scholar 

  10. Yamaguchi M, Makino R, Shimokawa N: The 5??end sequences and exon organization in rat regucalcin gene. Mol Cell Biochem 165: 145–150, 1996

    Google Scholar 

  11. Shimokawa N, Yamaguchi M: Calcium administration stimulates the expression of calcium-binding protein regucalcin mRNA in rat liver. FEBS Lett 305: 151–154, 1992

    Google Scholar 

  12. Isogai M, Yamaguchi M: Calcium administration increases calcium-binding protein regucalcin concentration in the liver of rats. Mol Cell Biochem 143: 53–58, 1995

    Google Scholar 

  13. Yamaguchi M, Kanayama Y, Shimokawa N: Expression of calcium-binding protein regucalcin mRNA in rat liver is stimulated by calcitonin: the hormonal effect is mediated through calcium. Mol Cell Biochem 136: 43–48, 1994

    Google Scholar 

  14. Yamaguchi M, Oishi K, Isogai M: Expression of hepatic calcium-binding protein regucalcin mRNA is elevated by refeeding of fasted rats: Involvement of glucose, insulin and calcium as stimulating factors. Mol Cell Biochem 142: 35–41, 1995

    Google Scholar 

  15. Yamaguchi M, Oishi K: 17 ?-Estradiol stimulates the expression of hepatic calcium-binding protein regucalcin mRNA in rats. Mol Cell Biochem 143: 137–141, 1995

    Google Scholar 

  16. Yamaguchi M, Kanayama Y: Enhanced expression of calcium-binding protein regucalcin mRNA in regenerating rat liver. J Cell Biochem 57: 185–190, 1995

    Google Scholar 

  17. Makino R, Yamaguchi M: Expression of calcium-binding protein regucalcin mRNA in hepatoma cells. Mol Cell Biochem 155: 85–90, 1996

    Google Scholar 

  18. Chomozynski P, Sacchi N: Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 162: 156–159, 1987

    Google Scholar 

  19. Sambrook J, Fritsch EF, Maniatis T: Northern Hybridization. In: Molecular Cloning (2nd edn), Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1989, pp 7.39–7.52

    Google Scholar 

  20. Yamaguchi M, Isogai M: Tissue concentration of calcium-binding protein regucalcin in rats by enzyme-linked immunoadsorbent assay. Mol Cell Biochem 122: 65–68, 1993

    Google Scholar 

  21. Wessendorf JHM, Garfinkel S, Zhan X, Brown S, Maciag T: Identification of a nuclear localization sequence within the structure of the human interleukin-1??precursor. J Biol Chem 268: 22100–22104, 1993

    Google Scholar 

  22. Kosaki A, Webster NJ: Effect of dexamethasone on the alternative splicing of the insulin receptor mRNA and insulin action in HepG2 hepatoma cells. J Biol Chem 268: 21990–21996, 1993

    Google Scholar 

  23. Yamaguchi M, Kanayama Y: Calcium-binding protein regucalcin inhibits deoxyribonucleic acid synthesis in the nuclei of regenerating rat liver. Mol Cell Biochem 162: 121–126, 1996

    Google Scholar 

  24. Pfyffer GE, Haemmerli G, Heizmann CW: Calcium-binding proteins in human carcinoma cell lines. Proc Natl Acad Sci USA 81: 6632–6636, 1984

    Google Scholar 

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Murata, T., Shinya, N. & Yamaguchi, M. Expression of calcium-binding protein regucalcin mRNA in the cloned human hepatoma cells (HegG2): Stimulation by insulin. Mol Cell Biochem 175, 163–168 (1997). https://doi.org/10.1023/A:1006844815743

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  • DOI: https://doi.org/10.1023/A:1006844815743

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