Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Water Extract of Allium sativum L. Induces Apoptosis in Human Leukemia U937 Cells through Reactive Oxygen Species Generation

마늘 열수 추출물의 활성산소종 생성을 통한 인체백혈병세포의 apoptosis 유발

  • Choi, Woo-Young (Department of Biochemistry, Dongeui University) ;
  • Chung, Kyung-Tae (Department of Laboratory Science Dongeui University) ;
  • Yoon, Tae-Kyung (Department of Environmental Engineering Major Dongeui University) ;
  • Choi, Byung-Tae (Department of Anatomy, Graduate School of Oriental Medicine, Pusan National University) ;
  • Lee, Yong-Tae (Department of Physiology, College of Oriental Medicine Dongeui University) ;
  • Lee, Won-Ho (Department of Biology, College of Natural Sciences, Pusan National University) ;
  • Ryu, Chung-Ho (Division of Applied Life Science, Institute of Agriculture and Life Science, Gyeongsang National University) ;
  • Choi, Yung-Hyun (Department of Biochemistry, Dongeui University)
  • 최우영 (동의대학교 한의과대학 생화학교실) ;
  • 정경태 (동의대학교 자연과학대학 임상병리학과) ;
  • 윤태경 (동의대학교 공과대학 환경공학과) ;
  • 최병태 (부산대학교 한의학전문대학원 해부학교실) ;
  • 이용태 (동의대학교 한의과대학 생리학교실) ;
  • 이원호 (부산대학교 자연과학대학 생물학과) ;
  • 류충호 (경상대학교 농업생명과학대학 응용생명공학과) ;
  • 최영현 (동의대학교 한의과대학 생화학교실)
  • Published : 2007.12.30
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Abstract

The health benefits of garlic (Allium sativum L.) are derived from a wide variety of components and from the different ways it is administered. The known health benefits of garlic include cardiovascular protective effects, stimulation of immune function, reduction of blood glucose level, protection against microbial, viral and fungal infections, as well as anticancer effects. In the present study, it was examined the effects of water extract of A. sativum (WEAS) on the growth of cultured human tumor cells in order to investigate its anti-proliferative mechanism. Treatment of WEAS to tumor cells resulted in the growth inhibition, especially in leukemia cells, which was associated with induction of G2/M arrest of the cell cycle and apoptosis. In order to further explore the critical events leading to apoptosis in WEAS-treated U937 human leukemia cells, the following effects of WEAS on components of the mitochondrial apoptotic pathway were examined: generation of reactive oxygen species (ROS), alteration of the mitochondrial membrane potential (MMP), and the expression changes of Bcl-2 and IAP family proteins. The cytotoxic effect of WEAS was mediated by its induction of apoptosis as characterized by the occurrence of DNA ladders, apoptotic bodies and chromosome condensation in U937 cells. The WEAS-induced apoptosis in U937 cells was correlated with the generation of intracellular ROS, collapse of MMP, activation of caspase-3 and down-regulation of anti-apoptotic proteins. The quenching of ROS generation with antioxidant N-acetyl-L-cysteine conferred significant protection against WEAS-elicited ROS generation, caspase-3 activation, G2/M arrest and apoptosis. In conclusion, the present study reveals that the cellular ROS generation plays a pivotal role in the initiation of WEAS-triggered apoptotic death in U937 cells.

본 연구에서는 몇 가지 암세포를 대상으로 마늘 열수추출물(WEAS)의 항암활성을 조사하였으며, 비교적 높은 감수성을 보인 백혈병세포를 대상으로 세포증식억제가 apoptosis 유도와 연관성이 있음을 제시하였다. WEAS에 의한 U937 세포의 apoptosis 유도는 세포주기 G2/M arrest와 연관성이 있었으며, 다양한 apoptosis조절 유전자들의 발현 변화를 동반하였음을 확인하였다. 이러한 apoptosis조절 인자들의 발현변화는 미토콘드리아 막 전위의 소실과 연관성이 있었으며, WEAS에 의한 암세포의 G2/M arrest에 의한 apoptosis 유발에 ROS가 중요한 조절자로 작용함을 알 수 있었다.

Keywords

References

  1. Antonsson, B. and J. C. Martinou. 2000. The Bcl-2 protein family. Exp. Cell Res. 256, 50-57 https://doi.org/10.1006/excr.2000.4839
  2. Arunkumar, A., M. R., Vijayababu, N. Srinivasan, M. M. Aruldhas and J. Arunakaran. 2006. Garlic compound, diallyl disulfide induces cell cycle arrest in prostate cancer cell line PC-3. Mol. Cell Biochem. 288, 107-113 https://doi.org/10.1007/s11010-006-9126-6
  3. Barisic, K., J. Petrik and L. Rumora. 2003. Biochemistry of apoptotic cell death. Acta. Pharm. 53, 151-164
  4. Borek, C. 2006. Garlic reduces dementia and heart-disease risk. J. Nutr. 136, 810S-812S https://doi.org/10.1093/jn/136.3.810S
  5. Chiarugi, V., L. Magnelli, M. Cinelli and G. Basi. 1994. Apoptosis and the cell cycle. Cell Mol. Biol. Res. 40, 603-612
  6. Das, A., N. L. Banik and S. K. Ray. 2007. Garlic compounds generate reactive oxygen species leading to activation of stress kinases and cysteine proteases for apoptosis in human glioblastoma T98G and U87MG cells. Cancer 110, 1083-1095 https://doi.org/10.1002/cncr.22888
  7. de Almeida, C. J. and R. Linden. 2005. Phagocytosis of apoptotic cells: a matter of balance. Cell Mol. Life Sci. 62, 1532-1546 https://doi.org/10.1007/s00018-005-4511-y
  8. de Martino, A., G. Filomeni, K. Aquilano,, M. R. Ciriolo and G. Rotilio. 2006. Effects of water garlic extracts on cell cycle and viability of HepG2 hepatoma cells. J. Nutr. Biochem. 17, 742-749 https://doi.org/10.1016/j.jnutbio.2005.12.005
  9. Frantz, D. J., B. G. Hughes, D. R. Nelson, B. K. Murray and M. J. Christensen. 2000. Cell cycle arrest and differential gene expression in HT-29 cells exposed to an aqueous garlic extract. Nutr. Cancer 38, 255-264 https://doi.org/10.1207/S15327914NC382_15
  10. Fruehauf, J. P. and F. L. Meyskens Jr. 2007. Reactive oxygen species: a breath of life or death? Clin. Cancer Res. 13, 789-794 https://doi.org/10.1158/1078-0432.CCR-06-2082
  11. Fulda, S. and K.M. Debatin. 2004. Apoptosis signaling in tumor therapy. Ann. N.Y. Acad. Sci. 1028, 150-156 https://doi.org/10.1196/annals.1322.016
  12. Herman-Antosiewicz, A., A. A. Powolny and S. V. Singh. 2007. Molecular targets of cancer chemoprevention by garlic-derived organosulfides. Acta. Pharmacol. Sin. 28, 1355-1364 https://doi.org/10.1111/j.1745-7254.2007.00682.x
  13. Holcik, M., H. Gibson and R. G. Korneluk. 2001. XIAP: apoptotic brake and promising therapeutic targe. Apoptosis 6, 253-261 https://doi.org/10.1023/A:1011379307472
  14. Hussein, M. R., A. K. Haemel and G. S. Wood. 2003. Apoptosis and melanoma: molecular mechanisms. J. Pathol. 199, 275-288 https://doi.org/10.1002/path.1300
  15. Jaeschke, H. and M. L. Bajt. 2006. Intracellular signaling mechanisms of acetaminophen-induced liver cell death. Toxicol. Sci. 89, 31-41 https://doi.org/10.1093/toxsci/kfi336
  16. Jakubikova, J. and J. Sedlak. 2006. Garlic-derived organosulfides induce cytotoxicity, apoptosis, cell cycle arrest and oxidative stress in human colon carcinoma cell lines. Neoplasma 53, 191-199
  17. Kaufmann, S. H., S. Desnoyers, Y. Ottaviano, N. E. Davidson and G. G. Poirier. 1993. Specific proteolytic cleavage of poly(ADP-ribose) polymerase: an early marker of chemotherapy-induced apoptosis. Cancer Res. 53, 3976-3985
  18. Khanum, F., K. R. Anilakumar and K. R. Viswanathan. 2004. Anticarcinogenic properties of garlic: a review. Crit. Rev. Food Sci. Nutr. 44, 479-488 https://doi.org/10.1080/10408690490886700
  19. Kim, R., M. Emi, K. Tanabe, S. Murakami, Y. Uchida and K. Arihiro. 2006. Regulation and interplay of apoptotic and non-apoptotic cell death. J. Pathol. 208, 319-326 https://doi.org/10.1002/path.1885
  20. Kwon, S. K. 2003. Organosulfur compounds from and physiological activities. J. App. Pharmacol. 11, 8-32
  21. Lazebnik, Y. A., S. H. Kaufmann, S. Desnoyers, G. G. Poirier and W. C. Earnshaw. 1994. Cleavage of poly (ADP-ribose) polymerase by a proteinase with properties like ICE. Nature 371, 346-347 https://doi.org/10.1038/371346a0
  22. Lin, H. L., J. S. Yang, J. H. Yang, S. S. Fan, W. C. Chang, Y. C. Li and J. G. Chung. 2006. The role of $Ca^{2+}$ on the DADS-induced apoptosis in mouse-rat hybrid retina ganglion cells (N18). Neurochem. Res. 31, 383-393 https://doi.org/10.1007/s11064-005-9035-1
  23. Lu, H. F., C. C. Sue, C. S. Yu, S. C. Chen, G. W. Chen and J. G. Chung. 2004. Diallyl disulfide (DADS) induced apoptosis undergo caspase-3 activity in human bladder cancer T24 cells. Food Chem. Toxicol. 42, 1543-1552 https://doi.org/10.1016/j.fct.2003.06.001
  24. Ngo, S. N., D.B. Williams, L. Cobiac and R. J. Head. 2007. Does garlic reduce risk of colorectal cancer? A systematic review. J. Nutr. 137, 2264-2269 https://doi.org/10.1093/jn/137.10.2264
  25. Pittler, M. H. and E. Ernst. 2007. Clinical effectiveness of garlic (Allium sativum). Mol. Nutr. Food Res. 51, 1382-1385 https://doi.org/10.1002/mnfr.200700073
  26. Ryter, S. W., H. P. Kim, A. Hoetzel, J. W. Park, K. Nakahira, X. Wang and A. M. Choi. 2007. Mechanisms of cell death in oxidative stress. Antioxid. Redox Signal. 9, 49-89 https://doi.org/10.1089/ars.2007.9.49
  27. Sabayan, B., F. Foroughinia and A. Chohedry. 2007. postulated role of garlic organosulfur compounds in prevention of valproic acid hepatotoxicity. Med. Hypotheses 68, 512-514 https://doi.org/10.1016/j.mehy.2006.07.055
  28. Salvesen, G. S. and C. S. Duckett. 2002. IAP proteins: blocking the road to death's door. Nat. Rev. Mol. Cell Biol. 3, 401-410 https://doi.org/10.1038/nrm830
  29. Schulze-Osthoff, K., D. Ferrari, M. Los, S. Wesselborg and M. E. Peter. 1998. Apoptosis signaling by death receptors. Eur. J. Biochem. 254, 439-459 https://doi.org/10.1046/j.1432-1327.1998.2540439.x
  30. Shukla, Y. and N. Kalra. 2007. Cancer chemoprevention with garlic and its constituents. Cancer Lett. 247, 167-181 https://doi.org/10.1016/j.canlet.2006.05.009
  31. Su, C. C., G. W. Chen, T. W. Tan, J. G. Lin and J. G. Chung. 2006. Crude extract of garlic induced caspase-3 gene expression leading to apoptosis in human colon cancer cells. In Vivo 20, 85-90
  32. Tattelman, E. 2005. Health effects of garlic. Am. Fam. Physician 72, 103-106
  33. Wu, X. J., F. Kassie and V. Mersch-Sundermann. 2005. The role of reactive oxygen species (ROS) production on diallyl disulfide (DADS) induced apoptosis and cell cycle arrest in human A549 lung carcinoma cells. Mutat. Res. 579, 115-124 https://doi.org/10.1016/j.mrfmmm.2005.02.026
  34. Wu, X. J., T. Stahl, Y. Hu, F. Kassie and V. Mersch-Sundermann. 2006. The production of reactive oxygen species and the mitochondrial membrane potential are modulated during onion oil-induced cell cycle arrest and apoptosis in A549 cells. J. Nutr. 136, 608-613 https://doi.org/10.1093/jn/136.3.608
  35. Wylie, A. H., J. F. Kerr and A. R. Currie. 1980. Cell death: the significance of apoptosis. Int. Rev. Cytol. 68, 251-306 https://doi.org/10.1016/S0074-7696(08)62312-8
  36. Xiao, D., J. T. Pinto, G. G. Gundersen and I. B. Weinstein. 2005. Effects of a series of organosulfur compounds on mitotic arrest and induction of apoptosis in colon cancer cells. Mol. Cancer Ther. 4, 1388-1398 https://doi.org/10.1158/1535-7163.MCT-05-0152