Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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The Effect of Overexpression of Rat Clusterin in L929 Fibroblasts

  • PARK , JUNG-HYUN (Experimental Immunology Branch, National Cancer Institute) ;
  • JU, SUNG-KYU (School of Bioscience and Biotechnology, Chungnam National University) ;
  • PARK, JEE-SUN (School of Bioscience and Biotechnology, Chungnam National University) ;
  • PARK, YOO-KYOUNG (Department of Food and Nutrition, Hannam University) ;
  • KANG, MYUNG-HEE (Department of Food and Nutrition, Hannam University) ;
  • YOU, KWAN-HEE (School of Bioscience and Biotechnology, Chungnam National University)
  • Published : 2004.12.01
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Abstract

Oxidants such as hydrogen peroxide are powerful inducers of cell damage, ageing, and apoptosis. Since clusterin, a 75-80 kDa mammalian glycoprotein, is frequently found to be inducible in apoptotic cells and tissues, this study inquired into whether this would be a protective mechanism against further cell death. The aim was to find out whether overexpression of clusterin could protect cells from oxidant­induced stress and apoptosis. To clarify this issue, we generated and analyzed stable cell lines expressing fusion proteins of a rat clusterin with an enhanced green fluorescent protein (EGFP). When treated with varying concentrations of hydrogen peroxides, clusterin transfectants indeed showed increased resistance to apoptosis and exhibited a much higher survival rate than mock-transfected cells. On the other hand, neither intracellular re-distribution nor local concentration of clusterin-EGFP was observed, which leaves the question open about its anti-apoptotic mechanism. In conclusion, the overexpression of clusterin provides a means for protecting cells against oxidative stress and subsequent cell death.

Keywords

References

  1. Bettuzzi, S., L. Troiano, P. Davalli, F. Troea, M. C. Ingletti, E. Grassilli, D. Monti, A. Corti, and C. Franceschi. 1991. In vivo accumulation of sulfated glycoprotein 2 mRNA in rat thymocytes upon dexamethasone-induced cell death. Biochem. Biophys. Res. Commun. 175: 810-815
  2. Buttyan, R. C., C. A. Olsson, J. Pintar, C. Chang, M. Bandyk, P. Y. Ng, and I. S. Sawczuk, 1989. Induction of the TRPM-2 gene in cells undergoing programmed death. Mol. Cell Biol. 9: 3473-3481
  3. Davies, K. J. A. 1995. Oxidative stress: The paradox of aerobic life. Biochem. Soc. Symp. 61: 1-31
  4. Davies, K. J. A. 1999. The broad spectrum of responses to oxidants in proliferating cells: A new paradigm for oxidative stress. IUBMB Life 48: 41-47
  5. Dumont, P., F. Chainiaux, F. Eliaers, C. Petropoulou, J. Remacle, C. Koch-Brandt, E. S. Gonos, and O. Toussaint. 2002. Overexpression of apolipoprotein J in human fibroblasts protects against cytotoxicity and premature senescence induced by ethanol and tert-butylhydroperoxide. Cell Stress Chaperones 7: 23-35
  6. Fritz, I. B., K. Burdzy, O. Setchell, and O. Blaschuk. 1983. Ram rete testis fluid contains a protein (clusterin) which influences cell-cell interactions in vitro. Biol. Reprod. 28: 1173-1188
  7. Han, B. H., R. B. DeMattos, L. L. Dugan, J. S. Kim-Han, R. P. Brendza, J. D. Fryer, M. Kierson, J. Cirrito, K. Quick, J. A. Harmony, B. J. Aronow, and D. M. Holtzman. 2001. Clusterin contributes to caspase-3-independent brain injury following neonatal hypoxia-ischemia. Nat. Med. 7: 338- 343
  8. Humphreys, D., T. T. Hochgrebe, S. B. Easterbrook-Smith, M. P. Tenniswood, and M. R. Wilson. 1997. Effects of clusterin overexpression on TNF-$\alpha$- and TGF-$\beta$-mediated death of L929 cells. Biochemistry 36: 15233-15243
  9. Jones, S. E. and C. Jomary. 2002. Clusterin. Int. J. Biochem. Cell Biol. 34: 427-431
  10. Ju, S. K., J. H. Park, S. Y. Na, K. H. You, K. L. Kim, and M. K. Lee. 2001. Determination of rat leptin activity in vitro using a novel luciferase reporter assay. Mol. Cells 12: 131- 136
  11. Kim, J. H., J. H. Park, Y. J. Lee, E. W. Cho, Y. S. Bae, and K. L. Kim. 2000. In vitro binding analysis of hepatitis B virus-preS-derived putative helper T-cell epitopes to MHC class II molecules using stable HLA-DRB1*0405/-DRA*0101 transfected cells. IUBMB Life 50: 379-384 https://doi.org/10.1080/713803746
  12. Montpetit, M. L., K. R. Lawless, and M. Tenniswood. 1986. Androgen-repressed messages in the rat ventral prostate. Prostate 8: 25-36
  13. Park, H. W., D. Y. Jun, and Y. H. Kim. 2002. Apoptotic activity of insect pathogenic fungus Paecilomyces japonica toward human acute leukemia jurkat T cells is associated with mitochondria-dependent caspase-3 activation regulated by Bcl-2. J. Microbiol. Biotechnol. 12: 950-956
  14. Park, J. H., S. Y. Na, H. H. Lee, Y. J. Lee, and K. L. Kim. 2001. Detection of pET-vector encoded, recombinant Stagged proteins using the monoclonal antibody ATOM-2. Hybridoma 20: 17-23
  15. Park, J. H., J. S. Park, S. K. Ju, K. B. Lee, Y. K. Park, M. H. Kang, S. Y. Na, and K. H. You. 2003. Clusterin mRNA expression in apoptotic and activated rat thymocytes. Cell Res. 13: 49-58
  16. Reddy, K. B., M. C. Karode, A. K. Harmony, and P. H. Howe. 1996. Interaction of transforming growth factor beta receptors with apolipoprotein J/clusterin. Biochemistry 35: 309-314.
  17. Reddy, K. B., G. Jin, M. C. Karode, J. A. Harmony, and P. H. Howe. 1996. Transforming growth factor beta (TGF- $\beta$)-induced nuclear localization of apolipoprotein J/clusterin in epithelial cells. Biochemistry 35: 6157-6163
  18. Schwochau, G. B., K. A. Nath, and M. E. Rosenberg. 1998. Clusterin protects against oxidative stress in vitro through aggregative and nonaggregative properties. Kidney Intl. 53: 1647-1653
  19. Song, J. S., S. B. Kim, Y. H. Lee, K. W. Lee, K. W. Lee, H. H. Jung, M. H. Kim, K. T. Kim, R. Brown, and Y. T. Kim. 2002. Adenovirus-mediated antisense expression of telomerase template RNA induces apoptosis in lung cancer cells. J. Microbiol. Biotechnol. 12: 89-95
  20. Viard, I., P. Wehrli, L. Jornot, R. Bullani, J. L. Vechietti, J. A. Schifferli, J. Tschopp, and L. E. French. 1999. Clusterin gene expression mediates resistance to apoptotic cell death induced by heat shock and oxidative stress. J. Invest. Dermatol. 112: 290-296
  21. Wilson, R. M. and S. B. Easterbrook-Smith. 2000. Clusterin is a secreted mammalian chaperone. Trends Biochem. Sci. 25: 95-98
  22. Yang, C. R., K. Leskov, K. Hosley-Eberlein, T. Criswell, J. J. Pink, T. J. Kinsella, and D. A. Boothman. 2000. Nuclear clusterin/XIP8, an X-ray-induced Ku70-binding protein that signals cell death. Proc. Natl. Acad. Sci. USA 97: 5907- 5912