Abstract
Cell death induced by menadione (vitamin K-3,2-methyl-1,4-naphthoquinone) has been investigated in human promyelocytic leukemia HL-60 cells. Menadione was found to induce both apoptosis and necrosis in HL-60 cells. Low concentration (1~50 µM) of menadione induced apoptotic cell death, which was demonstrated by typical DNA ladder patterns on agarose gel electrophoresis and flow cytometry analysis. In contrast, a high concentration of menadione (100 µM) induced necrotic cell death, which was demonstrated by DNA smear pattern in agarose gel electrophoresis. Necrotic cell death was accompanied with a great reduction of cell viability. Menadione activated caspase-3, as evidenced by both increased protease activity and proteolytic cleavage of 116 kDa poly(ADP-ribose) polymerase (PARP) into 85 kDa cleavage product. Caspase-3 activity was maximum at 50 µM of menadione, and very low at 100 µM of menadione. Taken together, our results showed that menadi-one induced mixed types of cell death, apoptosis at low concentrations and necrosis at high concentrations in HL-60 cells.
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Brown, P.C, Dulik, D.M and Jones, T.W (1991). The toxicity of menadione (2-methyl-1,4-naphthoquinone) and two thioether conjugates studied with isolated renal epithelial cells. Arch. Biochem. Biophys., 285, 187–196.
Chlebowski, R., Dietrich, M., Akman, S. and Block, J.B (1985). Vitamin K3 inhibition of malignant murine cell growth and human tumor colony formation. Cancer Treat. Rep., 69, 527–532.
Di Monte, D., Bellomo, G., Thor, H., Nicotera, P. and Orrenius, S. (1984). Menadione-induced cytotoxicity is associated with protein thiol oxidation and alteration in intracellular Ca2+ homeostasis. Arch. Biochem. Biophys., 235, 343–350.
Earnshaw, W.C, Martins, L.M and Kaufmann, S.H (1999). Mammalian caspases: structure, activation, substrates, and functions during apoptosis. Annu. Rev. Biochem., 68, 383–424.
Ferrer, I. and Planas, A.M (2003). Signaling of cell death and cell survival following focal cerebral ischemia: life and death struggle in the penumbra. J. Neuropathol. Exp. Neurol., 62, 329–339.
Hetts, S.W (1998). To die or not die: An overview of apopto-sis and its role in disease. J. AM. Med. Assoc., 279, 300–307.
Hengartner, M.O (2000). The biochemistry of apoptosis. Nature, 407, 770–776.
Hotti, A., Järvinen, K., Siivola, P. and Hölttä, E. (2000). Caspases and mitochondria in c-Myc-induced apoptosis: identification of ATM as a new target of caspases. Oncogene, 19, 2354–2362.
Janicke, R.U, Sprengart, M.L, Wati, M.R and Porter, A.G (1998). Caspase-3 is required for DNA fragmentation and morphological changes associated with apoptosis. J. Biol. Chem., 273, 9357–9360.
Kothakota, S., Azuma, T., Reinhard, C., Klippel, A., Tang, J., Chu, K., McGarry, T.J, Kirschner, M., Koths, K., Kwiat-kowski, D.J and Williams, L.T (1997). Caspase-3-gener-ated fragment of gelsolin: effector of morphological change in apoptosis. Science, 278, 294–298.
Los, M., Mozoluk, M., Ferrari, D., Stepczynska, A., Stroh, C., Renz, A., Herceq, Z., Wang, Z.Q and Schulze-Osthoff, K. (2002). Activation and caspase-mediated inhibition of PARP: a molecular switch between fibroblast necrosis and apoptosis in death receptor signaling. Mol. Biol. Cell, 13, 978–988.
Lippke, J., Gu, Y., Sarnecki, C., Caron, P.R and Su, M.S (1996). Identification and characterization of CPP32/Mch2 homolog 1, a novel cysteine protease similar to CPP32. J. Biol. Chem., 271, 1825–1828.
Majno, G. and Joris, I. (1995). Apoptosis, oncosis, and necrosis. An overview of cell death. Am. J. Pathol., 146, 3–15.
Margolin, A., Akman, S.A, Leong, L.A, Morgan, R., Somlo, G., Raschko, J.W, Ahn, C. and Doroshow, J.H (1995). Phase I study of mitomycin C and menadione in advanced solid tumors. Cancer Chemother. Pharmacol., 36, 293–298.
Morgan, W.A, Hartley, J.A and Cohen, G.M (1992). Quinone-induced DNA single strand breaks in rat hepatocytes and human chronic myelogenous leukaemic K562 cells. Bio-chem. Pharmacol., 44, 215–221.
Nagata, S., Nagase, H., Kawane, K., Mukae, N. and Fukuyama, H. (2003). Degradation of chromosomal DNA during apoptosis. Cell Death Differ., 10, 108–116.
Ngo, E.O, Sun, T.P, Chang, J.Y, Wang, C.C, Chi, K.H, Cheng, A.L and Nutter, L.M (1991). Menadione-induced DNA damage in a human tumor cell line. Biochem. Pharmacol., 42, 1961–1968.
Nicotera, P., McConkey, D., Svensson, S.A, Bellomo, G. and Orrenius, S. (1988). Correlation between cytosolic Ca+2 concentration and cytotoxicity in hepatocytes exposed to oxidative stress. Toxicology, 52, 55–63.
Packham, G., Ashmun, R.A and Cleveland, J.L (1996). Cytokines suppress apoptosis independent of increases in reactive oxygen levels. J. Immunol., 156, 2792–2800.
Scovassi, A.I and Poirier, G.G (1999). Poly (ADP-ribosyla-tion) and apoptosis. Mol. Cell. Biochem., 199, 125–137.
Stubberfield, C.R and Cohen, G.M (1988). NAD+ depletion and cytotoxicity in isolated hepatocytes. Biochem. Pharmacol., 37, 3967–3974.
Wyllie, A.H (1980). Glucocorticoid-induced thymocyte apopto-sis is associated with endogeneous endonuclease activation. Nature, 284, 555–556.
Yoo, B.S, Jung, K.H, Hana, S.B and Kim, H.M (1997). Apo-ptosis-mediated immunotoxicity of polychlorinated biphe-nyls (PCBs) in murine splenocytes. Toxicol. Lett., 91, 83–89.
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Sa, DJ., Lee, EJ. & Yoo, BS. Apoptosis Induction by Menadione in Human Promyelocytic Leukemia HL-60 Cells. Toxicol Res. 25, 113–118 (2009). https://doi.org/10.5487/TR.2009.25.3.113
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DOI: https://doi.org/10.5487/TR.2009.25.3.113