Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
권호 표지
DOI QR코드

DOI QR Code

Effect of Diallyl Disulfide on Heme Oxygenase-1 Expression in Human Hepatoma Cell Line HepG2

인간 간암세포주 HepG2에서 heme oxygenase-1 발현에 대한 diallyl disulfide의 효과

  • Kim, Kang-Mi (Department of Microbiology & Immunology, Pusan National University School of Medicine) ;
  • Lee, Sang-Kwon (Cardiovascular Center, Pusan National University Yangsan Hospital) ;
  • Park, Young-Chul (Department of Microbiology & Immunology, Pusan National University School of Medicine)
  • 김강미 (부산대학교 의학전문대학원 미생물학 및 면역학교실) ;
  • 이상권 (양산부산대학교병원 심혈관센터) ;
  • 박영철 (부산대학교 의학전문대학원 미생물학 및 면역학교실)
  • Received : 2011.06.10
  • Accepted : 2011.07.15
  • Published : 2011.07.30
Download PDF
⟨ Previous Next ⟩

Abstract

Diallyl disulfide (DADS), the most prevalent oil-soluble organosulfur compound in garlic, is known to have diverse biological activities, including anticarcinogenic, antiatherosclerotic, antiinflammatory, and antioxidant actions. In this study, we investigated the effect of DADS on the expression of heme oxygenase-1 (HO-1) in human liver hepatoma cell line HepG2. Treatment of HepG2 cells by DADS evoked a dose-dependent growth inhibition without significant toxicity to the cells, and also induced the expression of transcription factor Nrf2. However, DADS did not have any enhancing effect on transcription and translation of HO-1 expression in HepG2 cells. In addition, DADS efficiently blocked protein synthesis of HO-1 in HepG2 cells stimulated by CoPP or hemin. But, DADS did not decrease the content of transcripts of HO-1 gene stimulated by CoPP, with accumulation of Nrf2 and small Maf in the nucleus. Based on these results, we conclude that DADS inhibits HO-1 expression by modulation of translational level of CoPP or hemin-induced HO-1 expression in HepG2 cells.

Dially disulfide (DADS)는 마늘의 주요한 유기 황화합물 성분으로서 다양한 약리 작용을 나타낸다. 최근 DADS가 항염증과 항동맥경화 작용뿐만 아니라 암세포의 증식을 억제하고 사멸을 유도한다는 보고가 이어지고 있고, 이에 관련된 연구가 활발히 진행되고 있는 실정이다. 한편, DADS가 세포 내 항산화 인자인 glutathione을 증가시킨다는 연구결과와 세포 내 항산화 효소의 일종인 HO-1의 발현을 직접 유도한다는 결과가 보고되었다. 그래서, 본 연구에서는 논란이 되고 있는 DADS의 세포 내 항산화 효소인 HO-1의 발현에서의 효과 및 그 전사인자들의 작용에 관여하는지를 인간 간암세포주 HepG2에서 조사하였다. 배양 중인 HepG2 세포에서 DADS는 독성이 없는 농도에서 세포의 증식을 크게 억제하였고, 전사인자 Nrf2의 발현을 약하게 유도하였으나 HO-1의 발현에는 영향을 미치지 못하는 것으로 나타났다. 또한, DADS는 HO-1 유도제인 CoPP와 hemin에 의해 자극된 HepG2 세포의 HO-1 발현의 증가를 단백질 수준에서 강력하게 억제시키는 것으로 나타났다. 그러나 DADS는 CoPP에 의한 HO-1 유전자의 mRNA 수준의 전사에는 억제 효과를 보이지 않았으며, 또한 Nrf2와 small Maf의 발현을 증가시키고 핵 내에 축적시키는 것으로 나타났다. 이를 종합해 볼 때 DADS는 단독으로 HO-1 발현을 유도하지 못하고, HO-1 유도제에 의한 HO-1 유전자의 발현과정에서는 전사단계가 아닌 번역단계에서 역할을 함으로써 HO-1의 단백질 합성을 억제하는 것으로 보인다. 결론적으로, 항산화 효소인 HO-1의 활성은 외부 자극으로부터 세포를 보호하고 사멸에 저항하게 하는데, DADS는 인간 간암세포주 HepG2에서 이 효소의 발현을 억제함으로써 항암제 및 redox 변화에 따른 암세포주의 성장을 억제하고 세포사멸을 촉진시킬 수 있다고 여겨진다.

Keywords

References

  1. Belloir, C., V. Singh, C. Daurat, M. H. Siess, and A. M. Le Bon. 2006. Protective effects of garlic sulfur compounds against DNA damage induced by direct- and indirect-acting genotoxic agents in HepG2 cells. Food Chem. Toxicol. 44, 827-834. https://doi.org/10.1016/j.fct.2005.11.005
  2. Bose, C., J. Guo, L. Zimniak, S. K. Srivastava, S. P. Singh, P. Zimniak, and S. V. Singh. 2002. Critical role of allyl groups and disulfide chain in induction of Pi class glutathione transferase in mouse tissues in vivo by diallyl disulfide, a naturally occuring chemopreventive agent in garlic. Carcinogenesis 23, 1661-1665. https://doi.org/10.1093/carcin/23.10.1661
  3. Cellini, L., E. Di Campli, M. Masulli, S. Di Bartolomeo, and N. Allocati. 1996. Inhibition of helicobacter pylori by garlic extract (Allium sativum). FEMS Immunol. Med. Microbiol. 13, 273-277. https://doi.org/10.1111/j.1574-695X.1996.tb00251.x
  4. Chen, C., D. Pung, V. Leong, V. Hebbar, G. Shen, S. Nair, W. Li, and A. N. Kong. 2004. Induction of detoxifying enzymes by garlic organosulfur compounds through transcription factor Nrf2: effect of chemical structure and stress signals. Free Radic. Biol. Med. 37, 1578-1590. https://doi.org/10.1016/j.freeradbiomed.2004.07.021
  5. Dausch, J. G. and D. W. Nixon. 1990. Garlic: a review of its relationship to malignant disease. Prev. Med. 19, 346-361. https://doi.org/10.1016/0091-7435(90)90034-H
  6. Durante, W., M. H. Kroll, N. Christodoulides, K. J. Peyton, and A. I. Schafer. 1997 Nitric oxide induces heme oxygenase- 1 gene expression and carbon monoxide production in vascular smooth muscle cells. Circ. Res. 80, 557-564. https://doi.org/10.1161/01.RES.80.4.557
  7. Fang, J., H. Nakamura, and A. K. Iyer. 2007. Tumor-targeted induction of oxystress for cancer therapy. J. Drug Target 15, 475-486. https://doi.org/10.1080/10611860701498286
  8. Fang, J., T. Akaike, and H. Maeda. 2004. Antiapoptotic role of heme oxygenase (HO) and the potential of HO as a target in anticancer treatment. Apoptosis 9, 27-35. https://doi.org/10.1023/B:APPT.0000012119.83734.4e
  9. Filomeni, G., K. Aquilano, G. Rotilio, and M. R. Ciriolo. 2003. Reactive oxygen species-dependent c-Jun NH2-terminal kinase/c-Jun signaling cascade mediates neuroblastoma cell deathinduced by diallyl disulfide. Cancer Res. 63, 5940-5949.
  10. Fleischauer, A. T. and L. Arab. 2001. Garlic and cancer: a critical review of the epidemiologic literature. J. Nutr. 131, 1032S-1040S.
  11. Gong, P., B. Hu, and A. I. Cederbaum. 2004. Diallyl sulfide induces heme oxygenase-1 through MAPK pathway. Arch. Biochem. Biophys. 15, 252-260.
  12. Jo, H. J., J. D. Song, K. M. Kim, Y. H. Cho, K. H. Kim, and Y. C. Park. 2008. Diallyl disulfide induces reversible G2/M phase arrest on a p53-independent mechanism in human colon cancer HCT-116 cells. Oncol. Rep. 19, 275-280.
  13. John, F. R., T. M. Gregory, and P. Alvaro. 2007. Heme oxygenase-1 induction by NRF2 requires inactivation of the transcription repressor BACH1. Nucleic Acids Res. 35, 7074-7086. https://doi.org/10.1093/nar/gkm638
  14. Keyse, S. M. and R. M. Tyrrell. 1989. Heme oxygenase is the major 32-kDa stress protein induced in human skin fibroblast by UVA radiation, hydrogen peroxide, and sodium arsenite. Proc. Natl. Acad. Sci. USA 86, 99-103. https://doi.org/10.1073/pnas.86.1.99
  15. Kwon, K. B., S. J. Yoo, D. G. Ryu, J. Y. Yang, H. W. Rho, J. S Kim, J. W. Park, H. R. Kim, and B. H. Park. 2002. Induction of apoptosis by diallyl disulfide through activation of caspase-3 in human leukemia HL-60 cells. Biochem. Pharmacol. 63, 41-47. https://doi.org/10.1016/S0006-2952(01)00860-7
  16. Lee, P. J., B. H. Jiang, B. Y. Chin, N. V. Iyer, J. Alam, G. L. Semenza, and A. M. Choi. 1997. Hypoxia-inducible factot-1 mediates transcriptional activation of the heme oxygenase-1 gene in response to hypoxia. J. Biol. Chem. 272, 5375-5381. https://doi.org/10.1074/jbc.272.9.5375
  17. Lin, M. C., E. J. Wang, C. Patten, M. J. Lee, F. Xiao, K. R. Reuhl, and C. S. Yang. 1996. Protective effect of diallyl sulfone against acetaminophen-induced hepatotoxicity in mice. J. Biochem. Toxicol. 11, 11-20. https://doi.org/10.1002/(SICI)1522-7146(1996)11:1<11::AID-JBT2>3.0.CO;2-Y
  18. McMahon, M., K. Itoh, M. Yamamoto, and J. D. Haye. 2003. Keap1-dependent proteasomal degradation of transcription factor Nrf2 contributes to the negative regulation of antioxidant response element-driven gene expression. J. Biol. Chem. 278, 21592-21600. https://doi.org/10.1074/jbc.M300931200
  19. Munday, J. S., K. A. James, L. M. Fray, S. W. Kirkwood, and K. G. Thompson. 1999. Daily supplementation with aged garlic extract, but not raw garlic, protects low density lipoprotein against in vitro oxidation. Atherosclerosis 143, 399-404. https://doi.org/10.1016/S0021-9150(98)00293-7
  20. Nakagawa, H., K. Tsuta, K. Kiuchi, H. Senzaki, K. Tanaka, K. Hioki, and A. Tsubura. 2001. Growth inhibitory effects of diallyl disulfide on human breast cancer cell lines. Carcinogenesis 22, 891-897. https://doi.org/10.1093/carcin/22.6.891
  21. Shan, Y., R. W. Lambrecht, S. E. Donohue, and H. L. Bonkovsky. 2006. Role of Bach1 and Nrf2 in up-regulation of the heme oxygenase-1 gene by cobalt protoporphyrin. FASEB J. 20, 2651-2653. https://doi.org/10.1096/fj.06-6346fje
  22. Song, J. D., S. K. Lee, K. M. Kim, S. E. Park, S. J. Park, K. H. Kim, S. C. Ahn, and Y. C. Park. 2009. Molecular mechanism of diallyl disulfide in cell cycle arrest and apoptosis in HCT-116 colon cancer cells. J. Biochem. Mol. Toxicol. 23, 71-79. https://doi.org/10.1002/jbt.20266
  23. Sundarm, S. G. and J. A. Milner. 1996. Diallyl disulfide induces apoptosis of human colon tumor cells. Carcinogenesis 17, 669-673. https://doi.org/10.1093/carcin/17.4.669
  24. Taketani, S., H. Kohno, T. Yoshinaga, and R. Tokunaga. 1989. The human 32-kDa stress protein induced by exposure to arsenite and cadmium ions is heme oxygenase. FEBS Lett. 245, 173-176. https://doi.org/10.1016/0014-5793(89)80215-7
  25. Tenhunen, R., H. S. Marver, and R. Schmid. 1968. The enzymatic conversion of heme to bilirubin by microsomal heme oxygenase. Proc. Natl. Acad. Sci. USA 61, 748-755. https://doi.org/10.1073/pnas.61.2.748
  26. Ryter, S. W., L. E. Otterbein, D. Morse, and A. M. Choi. 2002. Heme oxygenase/carbon monoxide signaling pathways: regulation and functional significance. Mol. Cell Biochem. 234, 249-263. https://doi.org/10.1023/A:1015957026924
  27. Wu, C. C., L. Y. Sheen, H. W. Chen, S. J. Tsai, and C. K. Lii. 2001. Effects of organosulfur compounds from garlic oil on the antioxidation system in rat liver and red blood cells. Food Chem. Toxicol. 39, 563-569. https://doi.org/10.1016/S0278-6915(00)00171-X
  28. Wu, C. C., L. Y. Sheen, H. W. Chen, W. W. Kuo, S. J. Tsai, and C. K. Lii. 2002. Differential effects of garlic oil and its three major organosulfur components on the hepatic detoxification system in rats. J. Agric. Food Chem. 50, 378-383. https://doi.org/10.1021/jf010937z
  29. Yang, C. S., S. K. Chhabra, J. Y. Hong, and T. J. Smith. 2001. Mechanisms of inhibition of chemical toxicity and carcinogenesis by diallyl sulfide (DAS) and related compounds from garlic. J. Nutr. 131, 1041S-1045S.
  30. Yu, R., C. Chen, Y. Y. Mo, V. Hebbar, E. D. Owuor, T. H. Tan, and A. N. Kong. 2000. Activation of mitogen-activated protein kinase pathways induces antioxidant response element- mediated gene expression via a Nrf2-dependent mechanism. J. Biol. Chem. 275, 39907-39913. https://doi.org/10.1074/jbc.M004037200
  31. Zhang, D. D. and M. Hannink. 2003. Distinct cysteine residues in Keap1 are required for Keap1-dependent ubiquitination of Nrf2 and for stabilization of Nrf2 by chemopreventive agents and oxidative stress. Mol. Cell Biol. 23, 8137-8151. https://doi.org/10.1128/MCB.23.22.8137-8151.2003