Induction of apoptosis by a hexane extract of aged black garlic in the human leukemic U937 cells
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BACKGROUND/OBJECTIVES: In this study, the apoptogenic activity and mechanisms of cell death induced by hexane extractof aged black garlic (HEABG) were investigated in human leukemic U937 cells. MATERIALS/METHODS: Cytotoxicity was evaluated by MTT (3-(4, 5-dimethyl-thiazol-2-yl)-2, 5-diphenyl tetrazoliumbromide) assay. Apoptosis was detected using 4,6-diamidino-2-phenyllindile (DAPI) staining, agarose gel electrophoresis and flow cytometry. The protein levels were determined by Western blot analysis. Caspase activity was measured using a colorimetric assay. RESULTS: Exposure to HEABG was found to result in a concentration- and time-dependent growth inhibition by inductionof apoptosis, which was associated with an up-regulation of death receptor 4 and Fas legend, and an increase in the ratioof Bax/Bcl-2 protein expression. Apoptosis-inducing concentrations of HEABG induced the activation of caspase-9, an initiatorcaspase of the mitochodrial mediated intrinsic pathway, and caspase-3, accompanied by proteolytic degradation ofpoly(ADP-ribose)-polymerase. HEABG also induced apoptosis via a death receptor mediated extrinsic pathway by caspase-8activation, resulting in the truncation of Bid, and suggesting the existence of cross-talk between the extrinsic and intrinsicpathways. However, pre-treatment of U937 cells with the caspase-3 inhibitor, z-DEVD-fmk, significantly blocked the HEABG-inducedapoptosis of these cells, and increased the survival rate of HEABG-treated cells, confirming that HEABG-induced apoptosis ismediated through activation of caspase cascade. CONCLUSIONS: Based on the overall results, we suggest that HEABG reduces leukemic cell growth by inducing caspase-dependentapoptosis through both intrinsic and extrinsic pathways, implying its potential therapeutic value in the treatment of leukemia.
Abstract
BACKGROUND/OBJECTIVES: In this study, the apoptogenic activity and mechanisms of cell death induced by hexane extractof aged black garlic (HEABG) were investigated in human leukemic U937 cells. MATERIALS/METHODS: Cytotoxicity was evaluated by MTT (3-(4, 5-dimethyl-thiazol-2-yl)-2, 5-diphenyl tetrazoliumbromide) assay. Apoptosis was detected using 4,6-diamidino-2-phenyllindile (DAPI) staining, agarose gel electrophoresis and flow cytometry. The protein levels were determined by Western blot analysis. Caspase activity was measured using a colorimetric assay. RESULTS: Exposure to HEABG was found to result in a concentration- and time-dependent growth inhibition by inductionof apoptosis, which was associated with an up-regulation of death receptor 4 and Fas legend, and an increase in the ratioof Bax/Bcl-2 protein expression. Apoptosis-inducing concentrations of HEABG induced the activation of caspase-9, an initiatorcaspase of the mitochodrial mediated intrinsic pathway, and caspase-3, accompanied by proteolytic degradation ofpoly(ADP-ribose)-polymerase. HEABG also induced apoptosis via a death receptor mediated extrinsic pathway by caspase-8activation, resulting in the truncation of Bid, and suggesting the existence of cross-talk between the extrinsic and intrinsicpathways. However, pre-treatment of U937 cells with the caspase-3 inhibitor, z-DEVD-fmk, significantly blocked the HEABG-inducedapoptosis of these cells, and increased the survival rate of HEABG-treated cells, confirming that HEABG-induced apoptosis ismediated through activation of caspase cascade. CONCLUSIONS: Based on the overall results, we suggest that HEABG reduces leukemic cell growth by inducing caspase-dependentapoptosis through both intrinsic and extrinsic pathways, implying its potential therapeutic value in the treatment of leukemia.
참고문헌 (25)
- Abramson N, "Leukocytosis: basics of clinical assessment", Am Fam Physician, 62, pp.2053~2060 (2000)
- Gilliland DG, "The molecular basis of leukemia", Hematology Am Soc Hematol Educ Program, pp.80~97 (2004)
- Voliotis D, "Challenges in treating hematologic malignancies", Semin Oncol, 29, pp.30~39 (2002)
- Leone G, "Therapy-related myeloid neoplasms", Curr Opin Oncol, 23, pp.672~680 (2011)
- Cotter TG, "Apoptosis and cancer: the genesis of a research field", Nat Rev Cancer, 9, pp.501~507 (2009)
- Jin Z, "Overview of cell death signaling pathways", Cancer Biol Ther, 4, pp.139~163 (2005)
- Elmore S, "Apoptosis: a review of programmed cell death", Toxicol Pathol, 35, pp.495~516 (2007)
- Lazebnik YA, "Cleavage of poly(ADP-ribose) polymerase by a proteinase with properties like ICE", Nature, 371, pp.346~347 (1994)
- Kroemer G, "Mitochondrial control of cell death", Nat Med, 6, pp.513~519 (2000)
- Kutuk O, "Regulation of Bcl-2 family proteins by posttranslational modifications", Curr Mol Med, 8, pp.102~118 (2008)
- Eun-Woo Lee, "The roles of FADD in extrinsic apoptosis and necroptosis", BMB Reports, 45(9), pp.496~508 (2012), 생화학분자생물학회
- Ashkenazi A, "Death receptors: signaling and modulation", Science, 281, pp.1305~1308 (1998)
- Lee EN, "Chloroform extract of aged black garlic attenuates TNF-α-induced ROS generation, VCAM-1 expression, NF-κB activation and adhesiveness for monocytes in human umbilical vein endothelial cells", Phytother Res, 25, pp.92~100 (2011)
- Dolara P, "Antioxidant vitamins and mineral supplementation, life span expansion and cancer incidence: a critical commentary", Eur J Nutr, 51, pp.769~781 (2012)
- : a review of its potential use as an anti-cancer agent, "Curr Cancer Drug Targets", 2003, 67~81
- Butt MS, "Garlic: nature's protection against physiological threats", Crit Rev Food Sci Nutr, 49, pp.538~551 (2009)
- Imai J, "Antioxidant and radical scavenging effects of aged garlic extract and its constituents", Planta Med, 60, pp.417~420 (1994)
- 이영민, "Antioxidant effect of garlic and aged black garlic in animal model of type 2 diabetes mellitus", Nutrition Research and Practice, 3(2), pp.156~161 (2009), 한국영양학회
- Kim HK, "5-Hydroxymethylfurfural from black garlic extract prevents TNFα-induced monocytic cell adhesion to HUVECs by suppression of vascular cell adhesion molecule-1 expression, reactive oxygen species generation and NF-κB activation", Phytother Res, 25, pp.965~974 (2011)
- Jeong YY, "Aged Red Garlic Extract Reduces Cigarette Smoke Extract-induced Cell Death in Human Bronchial Smooth Muscle Cells by Increasing Intracellular Glutathione Levels", PHYTOTHERAPY RESEARCH, 26(1), pp.18~25 (2012), WILEY-BLACKWELL
- Colín-González AL, "The antioxidant mechanisms underlying the aged garlic extract- and S-allylcysteine-induced protection", Oxid Med Cell Longev, 2012, pp.907162~ (2012)
- Wang X, "Aged black garlic extract induces inhibition of gastric cancer cell growth in vitro and in vivo", Mol Med Rep, 5, pp.66~72 (2012)
- Kim KH, "Hexane extract of aged black garlic reduces cell proliferation and attenuates the expression of ICAM-1 and VCAM-1 in TNF-α-activated human endometrial stromal cells", Int J Mol Med, 32, pp.67~78 (2013)
- Hongxi Zhao, "Triptolide inhibits ovarian cancer cell invasion by repression of matrix metalloproteinase 7 and 19 and upregulation of E-cadherin", Experimental and Molecular Medicine, 44(11), pp.633~641 (2012), 생화학분자생물학회
- Zhaohui Li, "Induction of apoptosis in HT-29 cells by quercetin through mitochondria-mediated apoptotic pathway", Animal Cells and Systems, 17(3), pp.147~153 (2013), 한국통합생물학회