Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Anti-apoptotic Activity of Heme Oxygenase-1 Up-regulated by Etoposide in Human Retinal Pigment Epithelial Cells

Etoposide에 의한 인간 망막색소상피세포인 ARPE-19 세포의 아폽토시스 과정에서 Heme oxygenase-1의 항아폽토시스 기능에 대한 연구

  • Lee, Sang-Kwon (Department of Cardiovascular Surgery and Medical Research Institute, Pusan National University School of Medicine) ;
  • Song, Ju-Dong (Department of Microbiology & Immunology, Pusan National University School of Medicine) ;
  • Kim, Kang-Mi (Department of Microbiology & Immunology, Pusan National University School of Medicine) ;
  • Kim, Jong-Min (Department of Anatomy and Cell Biology, Dong-A University School of Medicine) ;
  • Lee, Sang-Yull (Department of Biochemistry, Pusan National University School of Medicine) ;
  • Yoo, Young-Hyun (Department of Anatomy and Cell Biology, Dong-A University School of Medicine) ;
  • Park, Young-Chul (Department of Microbiology & Immunology, Pusan National University School of Medicine)
  • 이상권 (부산대학교 의학전문대학원 흉부외과학교실 및 의학연구소) ;
  • 송주동 (부산대학교 의학전문대학원 미생물학 및 면역학교실) ;
  • 김강미 (부산대학교 의학전문대학원 미생물학 및 면역학교실) ;
  • 김종민 (동아대학교 의과대학 해부학교실) ;
  • 이상률 (부산대학교 의학전문대학원 생화학교실) ;
  • 유영현 (동아대학교 의과대학 해부학교실) ;
  • 박영철 (부산대학교 의학전문대학원 미생물학 및 면역학교실)
  • Published : 2007.09.30
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Abstract

The topoisomerase II inhibitor etoposide causes an accumulation of DNA double strand breaks within the nuclei of cells. In this study, we investigated the effect of etoposide on the cell growth and apoptosis of human RPE cells. Etoposide evoked a significant inhibition of cell growth, and also induced DNA fragmentation in ARPE-19 cells. In addition, etoposide significantly up-regulated the expression of heme oxygenase-1 (HO-1), which is a stress-responsive protein and is known to play a protective role against the oxidative injury. And, etoposide-induced HO-1 expression was affected by the ROS scavenger N-acetyl cysteine. We also used oligonucleotides interfering with HO-1 mRNA (siRNA) for the inhibition of HO-1 expression. Interestingly, knock-down of the HO-1 gene significantly increased the level of DNA fragmentation in etoposide-treated ARPE-19 cells. In conclusion, these results suggest that up-regulated HO-1 plays as an anti-apoptotic factor in the process of apoptosis of ARPE-19 cells stimulated by etoposide.

Totopoisomerase II 저해제인 etoposide는 핵안에 DNA double strand breaks를 일으키므로써 세포의 DNA에 손상을 초래한다. 본 연구에서는 인간 망막색소상피세포인 ARPE-19 세포에서의 세포성장 및 아폽토시스에서 etoposide의 역할을 살펴보았다. Etoposide는 세포의 성장을 크게 감소시켰으며 TUNEL에서 아폽토시스를 나타내는 DNA fragmentation의 증가를 유도하였다. 게다가, etoposide는 산화적 손상에 대해 세포나 조직을 보호하는 역할을 하는 것으로 알려진 세포내 항산화효소인 heme oxygenase-1 (HO-1)의 발현을 크게 증가시켰다. Etoposide에 의한 HO-1 발현증가는 항산화물질 NAC에 의해 억제되었는데, 이는 etoposide에 의한 세포내 ROS의 증가가 HO-1 발현에 중요한 역할을 한다는 것을 의미한다. 또한 HO-1 발현을 억제하기 위하여 HO-1 siRNA 방법을 사용하였다. 흥미롭게도, HO-1 유전자의 knock-down은 etoposide에 의해 유도되는 DNA fragmentation의 정도를 증가시켰다. 이들 결과를 종합해볼 때, etoposide에 의해 자극되어진 ARPE-19 세포에서 발현증가된 HO-1은 etoposide에 의한 아폽토시스 유발과정에서 세포를 보호하는 항아폽토시스의 기능을 한다는 것을 시사한다.

Keywords

References

  1. Alizadeh, M., M. Wada, C. M. Celfman, J. T. Handa and L. M. Hjelmeland. 2001. Downregulation of differentiation specific gene expression by oxidative stress in ARPE-19 cells. Invest. Ophthalmol. Vis. Sci. 42, 2706-2713
  2. Angkeow, P., S. S. Deshpande, B. Qi, Y. X. Liu, Y. C. Park, B. H. Jeon, M. Ozaki and K. Irani. 2002. Redox factor-1: an extra-nuclear role in the regulation of endothelial oxidative stress and apoptosis. Cell Death Differ. 9, 717-725 https://doi.org/10.1038/sj.cdd.4401025
  3. Applegate, L. A, P. Luscher and R. M. Tyrrell. 1991. Induction of heme oxygenase: a general response to oxidant stress in cultured mammalian cells. Cancer Res. 51, 974-978
  4. Bok, D. 1993. The retinal pigment epithelium: a versatile partner in vision. J. Cell Sci. Suppl. 17, 189-195
  5. Burden, D. A and N. Osheroff. 1998. Mechanism of action of eukaryotic topoisomerase II and drugs targeted to the enzyme. Biochem. Biophys. Acta 1400, 139-154 https://doi.org/10.1016/S0167-4781(98)00132-8
  6. Choi, B. M., H. O. Pae, Y. R. Jeong, G. S. Oh, C. D. Jun, B. R. Kim, Y. M. Kim and H. T. Chung. 2004. Overexpression of heme oxygenase (HO)-1 renders Jurkat T cells resistant to Fas-mediated apoptosis: involvement of iron released by HO-1. Free Radic. Biol. Med. 36, 858-871 https://doi.org/10.1016/j.freeradbiomed.2004.01.004
  7. Detrick, B., D. A. Newsome, C. M. Percopo and J. J. Hooks. 1985. Class II antigen expression and gamma interferon modulation of monocytes and retinal pigment epithelial cells from patients with retinitis pigmentosa. Clin. Immunol. Immunopathol. 36, 201-211 https://doi.org/10.1016/0090-1229(85)90121-7
  8. Duckers, H. J., M. Boehm, A. L. True, S. F. Yet, H. San, J. L. Park, W. R. Clinton, M. E. Lee, G. J. Nabel and E. G. Nabel. 2001. Heme oxygenase-1 protects against vascular constriction and proliferation. Nat. Med. 7, 693-698 https://doi.org/10.1038/89068
  9. Elner, S. G., R. M. Strieter, V. M. Elner, B. J. Rollins, M. A. Del Monte and S. L. Kunkel. 1991. Monocyte chemotactic protein gene expression by cytokine-treated human retinal pigment epithelial cells. Lab Invest. 64, 819-825
  10. Elner, S. G., V. M. Elner, M. A. Pavilack, R. F. 3rd Todd, L. Mayo-Bond, W. A. Franklin, R. M. Strieter, S. L. Kunkel and A. R. Huber. 1992. Modulation and function of intercellular adhesion molecule-1 (CD54) on human retinal pigment epithelial cells. Lab Invest. 66, 200-211
  11. Elner, V. M., R. M. Strieter, S. G. Elner, M. Baggiolini, I. Lindley and S. L. Kunkel. 1990. Neutrophil chemotactic factor (IL-8) gene expression by cytokine-treated retinal pigment epithelial cells. Am. J. Pathol. 136, 745-750
  12. Gao, H. and J. G. Hollyfield. 1992. Aging of the human retina. Differential loss of neurons and retinal pigment epithelial cells. Invest. Ophthalmol. Vis. Sci. 33, 1-17
  13. Griffith, T. S., T. Brunner, S. M. Fletcher, D. R. Green and T. A. Ferguson. 1995. Fas ligand-induced apoptosis as a mechanism of immune privilege. Science 17, 1189-1192
  14. Hande, K. R. 1998. Etoposide: four decades of development of a topoisomerase II inhibitor. Eur. J. Cancer 34, 1514-1521 https://doi.org/10.1016/S0959-8049(98)00228-7
  15. Honda, S., L. M. Hjelmeland and J. T. Handa. 2000. The use of hyperoxia to induce chronic mild oxidative stress in RPE cells in vitro. Mol. Vis. 7, 63-70
  16. Hueber, A, P. Esser, K. Heimann, N. Kociok, S. Winter and M. Weller. 1998. The topoisomerase I inhibitors, camptothecin and beta-Iapachone, induce apoptosis of human retinal pigment epithelial cells. Exp. Eye Res. 67, 525-530 https://doi.org/10.1006/exer.1998.0544
  17. Hunt, R. C., D. M. Hunt, N. Gaur and A. Smith. 1996. Hemopexin in the human retina: protection of the retina against heme-mediated toxicity. J. Cell Physiol. 168, 71-80 https://doi.org/10.1002/(SICI)1097-4652(199607)168:1<71::AID-JCP9>3.0.CO;2-5
  18. Immenshuh, S. and G. Ramadori. 2000. Gene regulation of heme oxygenase-1 as a therapeutic target. Biochem. Pharmacol. 60, 1121-1128 https://doi.org/10.1016/S0006-2952(00)00443-3
  19. Jaffe, G. J., Le. L. Van, F. Valea, S. Haskill, W. Roberts, W. P. Arend, A. Stuart and W. P. Peters. 1992. Expression of interleukin-1 alpha, interleukin-1 beta, and an interleukin-1 receptor antagonist in human retinal pigment epithelial cells. Exp. Eye Res. 55, 325-335 https://doi.org/10.1016/0014-4835(92)90197-Z
  20. Jaffe, G. J., W. L. Roberts, H. L. Wong, A. D. Yurochko and C. J. Cianciolo. 1995. Monocyte-induced cytokine expression in cultured human retinal pigment epithelial cells. Exp. Eye Res. 60, 533-543 https://doi.org/10.1016/S0014-4835(05)80068-5
  21. Kutty, R. K., G. Kutty, B. Wiggert, C. J. Chader, R. M. Darrow and D. T. Organisciak. 1995. Induction of heme oxygenase 1 in the retina by intense visible light: suppression by the antioxidant dimethylthiourea. Proc. Natl. Acad. Sci. USA 92, 1177-1181 https://doi.org/10.1073/pnas.92.4.1177
  22. Liles, M. R., D. A. Newsome and P. D. Oliver. 1991. Antioxidant enzymes in the aging human retinal pigment epithelium. Arch. Ophthal. 109, 1285-1288 https://doi.org/10.1001/archopht.1991.01080090111033
  23. Maines, M. D. 1997. The heme oxygenase system: a regulator of second messenger gases. Annu. Rev. Pharmacol. Taxicol. 37, 517-554 https://doi.org/10.1146/annurev.pharmtox.37.1.517
  24. Mizumoto, K., R. J. Rothman and J. L. Farber. 1994. Programmed cell death (apoptosis) of mouse fibroblasts is induced by the topoisomerase II inhibitor etoposide. Mol. Pharmacol. 46, 890-895
  25. Otterbein, L. E. and A. M. K. Choi. 2000. Heme oxygenase: colors of defense against cellular stress. Am. J. Physiol. 279, L1029-L1037
  26. Park, S. E., J. D. Song, K. M. Kim, Y. M. Park, Y. H. Yoo and Y. C. Park. 2007. Diphenyleneiodonium induces ROS-independent p53 expression and apoptosis in human RPE cells. FEBS Lett. 581, 180-186 https://doi.org/10.1016/j.febslet.2006.12.006
  27. Planck, S. R., T. T. Dang, D. Craves, D. Tara, J. C. Ansel and J. T. Rosenbaum. 1992. Retinal pigment epithelial cells secrete interleukin-6 in response to interleukin-1. Invest. Ophthalmol. Vis. Sci. 33, 78-82
  28. Planck, S. R., X. N. Huang, J. E. Robertson and J. T. Rosenbaum. 1993. Retinal pigment epithelial cells produce interleukin-1 beta and granulocyte-macrophage colony-stimulating factor in response to interleukin-1 alpha. Curr. Eye Res. 12, 205-212 https://doi.org/10.3109/02713689308999465
  29. Platts, K. E., M. T. Benson, I. G. Rennie, R. M. Sharrard and R. C. Rees. 1995. Cytokine modulation of adhesion molecule expression on human retinal pigment epithelial cells. Invest. Ophthalmol. Vis. Sci. 36, 2262-2269
  30. Poss, K. D. and S. Tonegawa. 1997. Reduced stress defense in heme oxygenase-1-deficient cells. Proc. Natl. Acad. Sci. USA 94, 10925-10930 https://doi.org/10.1073/pnas.94.20.10925
  31. Schwartzman, M. L., J. Masferrer, M. W. Dunn, J. C. McGiff and N. G. Abraham. 1987. Cytochrome P450, drug metabolizing enzymes and arachidonic acid metabolism in bovine ocular tissues. Curr. Eye Res. 6, 623-630 https://doi.org/10.3109/02713688709025223
  32. Steinhorst, U. H., E. P. Chen, D. L. Hatchell, C. P. Samsa, P. Y. Saloupis, J. Westendorf and R. Machemer. 1993. Aclacinomycin A in the treatment of experimental proliferative vitreoretinopathy. Efficacy and toxicity in the rabbit eye. Invest. Ophthalmol. Vis. Sci. 34, 1753-1760
  33. Tate, D. J., D. A. Newsome and P. D. Oliver. 1993. Metallothionein shows an age-related decrease in human macular retinal pigment epithelium. Invest. Ophthalmol. Vis. Sci. 34, 2348-2351
  34. Tate, D. J., M. V. Miceli and D. A. Newsome. 1995. Phagocytosis and hydrogen peroxide induce catalase and metallothionein gene expression in human retinal pigment epithelial cells. Invest. Ophthalmol. Vis. Sci. 36, 1271-1279
  35. Varma, S. D. and R. D. Richards. 1988. Visual disorders and oxidative reactions. pp. 71-88, In Chow, C. K. (eds), Cellular antioxidant defense mechanisms, CRC Press, Boca Raton
  36. Velikay, M., U. Stolba, A. Wedrich, P. Datlinger, J. Akramian and S. Binder. 1994. The antiproliferative effect of fractionized radiation therapy: optimization of dosage. Doc. Ophthalmol. 87, 265-269 https://doi.org/10.1007/BF01203855
  37. Wiegund, R. D., J. G. Jose, L. M. Rapp and R. E. Anderson. 1984. Free radicals and damage to ocular tissues. pp. 317-353, In Armstrong, D., R. S. Sohal, R. C. Cutler and T. F. Slater (eds), Free radicals in molecular biology, aging, and disease, Raven Press, New York
  38. Wu, W. C, Y. H. Kao and D. N. Hu. 2002. A comparative study of effects of antiproliferative drugs on human retinal pigment epithelial cells in vitro. J. Ocul. Pharmacol. Ther. 18, 251-264 https://doi.org/10.1089/108076802760116179
  39. Yoon, H. S., J. H. Rho, K. W. Yoo, W. C. Park, S. H. Rho, Y. H. Choi, H. Suh, N. D. Kim, K. S. Yoo and Y. H. Yoo. 2001. Synthetic bile acid derivatives induce nonapoptotic death of human retinal pigment epithelial cells. Curr. Eye Res. 22, 367-374 https://doi.org/10.1076/ceyr.22.5.367.5499
  40. Yoon, H. S., S. C. Moon, N. D. Kim, B. S. Park and Y. H. Yoo. 2000. Genistein induces apoptosis of RPE-J cells by opening mitochondrial PTP. Biochem. Biophys. Res. Commun. 276, 151-156 https://doi.org/10.1006/bbrc.2000.3445
  41. Zinn, K. and J. V. Benjamin-Henkind. 1979. Anatomy of the human retinal pigment epithelium, pp. 3-31, In Zinn, K. M. and M. Marmor (eds), The retinal pigment epithelium, Harvard University Press, Cambridge