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


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.


  1. Antonsson, B. and J. C. Martinou. 2000. The Bcl-2 protein family. Exp. Cell Res. 256, 50-57
  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
  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
  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
  7. de Almeida, C. J. and R. Linden. 2005. Phagocytosis of apoptotic cells: a matter of balance. Cell Mol. Life Sci. 62, 1532-1546
  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
  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
  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
  11. Fulda, S. and K.M. Debatin. 2004. Apoptosis signaling in tumor therapy. Ann. N.Y. Acad. Sci. 1028, 150-156
  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
  13. Holcik, M., H. Gibson and R. G. Korneluk. 2001. XIAP: apoptotic brake and promising therapeutic targe. Apoptosis 6, 253-261
  14. Hussein, M. R., A. K. Haemel and G. S. Wood. 2003. Apoptosis and melanoma: molecular mechanisms. J. Pathol. 199, 275-288
  15. Jaeschke, H. and M. L. Bajt. 2006. Intracellular signaling mechanisms of acetaminophen-induced liver cell death. Toxicol. Sci. 89, 31-41
  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
  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
  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
  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
  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
  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
  25. Pittler, M. H. and E. Ernst. 2007. Clinical effectiveness of garlic (Allium sativum). Mol. Nutr. Food Res. 51, 1382-1385
  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
  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
  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
  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
  30. Shukla, Y. and N. Kalra. 2007. Cancer chemoprevention with garlic and its constituents. Cancer Lett. 247, 167-181
  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
  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
  35. Wylie, A. H., J. F. Kerr and A. R. Currie. 1980. Cell death: the significance of apoptosis. Int. Rev. Cytol. 68, 251-306
  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