Abstract
Cryptotanshinone is a biologically active compound from the root of Salvia miltiorrhiza. In the present study, we investigated the molecular mechanisms by which cryptotanshinone is in synergy with tumor necrosis factor-alpha (TNF-α) for the induction of apoptosis in human chronic myeloid leukemia (CML) KBM-5 cells. The co-treatment of cryptotanshinone with TNF-α reduced the viability of the cells [combination index (CI) < 1]. Concomitantly, the co-treatment of cryptotanshinone and TNF-α elicited apoptosis, manifested by enhanced the number of terminal deoxynucleotide transferase-mediated dUTP-nick-end labeling (TUNEL)-positive cells, the sub-G1 cell populations, and the activation of caspase-8 and -3, in comparison with the treatment with either drug alone. The treatment with cryptotanshinone further suppressed TNF-α-mediated expression of c-FLIPL, Bcl-xL, but the increased level of tBid (a caspase-8 substrate). Furthermore, cryptotanshinone activated p38 but not NF-κB in TNF-α-treated KBM-5 cells. The addition of a specific p38 MAPK inhibitor SB203580 significantly attenuated cryptotanshinone/TNF-α-induced apoptosis. The combination treatment of cryptotanshinone and TNF-α also stimulated the reactive oxygen species (ROS) generation. N-acetyl-l-cysteine (NAC, a ROS scavenger) was not only able to block cryptotanshinone/TNF-α-induced ROS production but also the activation of caspase-8 and p38 MAPK. Overall, our findings suggest that cryptotanshinone can sensitize TNF-α-induced apoptosis in human myeloid leukemia KBM-5 cells, which appears through ROS-dependent activation of caspase-8 and p38.
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References
Chan KF, Siegel MR, Lenardo JM (2000) Signaling by the TNF receptor superfamily and T cell homeostasis. Immunity 13:419–422
Van Herreweghe F, Festjens N, Declercq W, Vandenabeele P (2010) Tumor necrosis factor-mediated cell death: to break or to burst, that’s the question. Cell Mol Life Sci 67:1567–1579
Rath PC, Aggarwal BB (1999) TNF-induced signaling in apoptosis. J Clin Immunol 19:350–364
Locksley RM, Killeen N, Lenardo MJ (2001) The TNF and TNF receptor superfamilies: integrating mammalian biology. Cell 104:487–501
Jiang Y, Woronicz JD, Liu W, Goeddel DV (1999) Prevention of constitutive TNF receptor 1 signaling by silencer of death domains. Science 283:543–546
Cho KH, Shin SY, Lee HW, Wolkenhauer O (2003) Investigations into the analysis and modeling of the TNF alpha-mediated NF-kappa B-signaling pathway. Genome Res 13:2413–2422
Vietor I, Schwenger P, Li W, Schlessinger J, Vilcek J (1993) Tumor necrosis factor-induced activation and increased tyrosine phosphorylation of mitogen-activated protein (MAP) kinase in human fibroblasts. J Biol Chem 268:18994–18999
Antosiewicz J, Ziolkowski W, Kaczor JJ, Herman-Antosiewicz A (2007) Tumor necrosis factor-alpha-induced reactive oxygen species formation is mediated by JNK1-dependent ferritin degradation and elevation of labile iron pool. Free Radic Biol Med 43:265–270
Izeradjene K, Douglas L, Tillman DM, Delaney AB, Houghton JA (2005) Reactive oxygen species regulate caspase activation in tumor necrosis factor-related apoptosis-inducing ligand-resistant human colon carcinoma cell lines. Cancer Res 65:7436–7445
Woo CH, Eom YW, Yoo MH et al (2000) Tumor necrosis factor-alpha generates reactive oxygen species via a cytosolic phospholipase A2-linked cascade. J Biol Chem 275:32357–32362
Wang Z, Kishimoto H, Bhat-Nakshatri P, Crean C, Nakshatri H (2005) TNFalpha resistance in MCF-7 breast cancer cells is associated with altered subcellular localization of p21CIP1 and p27KIP1. Cell Death Differ 12:98–100
Tomek S, Horak P, Pribill I et al (2004) Resistance to TRAIL-induced apoptosis in ovarian cancer cell lines is overcome by co-treatment with cytotoxic drugs. Gynecol Oncol 94:107–114
Khwaja A, Tatton L (1999) Resistance to the cytotoxic effects of tumor necrosis factor alpha can be overcome by inhibition of a FADD/caspase-dependent signaling pathway. J Biol Chem 274:36817–36823
Kim H, Kim EH, Eom YW et al (2006) Sulforaphane sensitizes tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-resistant hepatoma cells to TRAIL-induced apoptosis through reactive oxygen species-mediated up-regulation of DR5. Cancer Res 66:1740–1750
Moon DO, Kim MO, Lee JD, Choi YH, Kim GY (2010) Rosmarinic acid sensitizes cell death through suppression of TNF-alpha-induced NF-kappaB activation and ROS generation in human leukemia U937 cells. Cancer Lett 288:183–191
Ahn KS, Sethi G, Aggarwal BB (2007) Simvastatin potentiates TNF-alpha-induced apoptosis through the down-regulation of NF-kappaB-dependent antiapoptotic gene products: role of IkappaBalpha kinase and TGF-beta-activated kinase-1. J Immunol 178:2507–2516
Kim EJ, Jung SN, Son KH et al (2007) Antidiabetes and antiobesity effect of cryptotanshinone via activation of AMP-activated protein kinase. Mol Pharmacol 72:62–72
Chen W, Luo Y, Liu L et al (2010) Cryptotanshinone inhibits cancer cell proliferation by suppressing mammalian target of rapamycin-mediated cyclin D1 expression and Rb phosphorylation. Cancer Prev Res (Phila Pa) 3:1015–1025
Park IJ, Kim MJ, Park OJ et al (2010) Cryptotanshinone sensitizes DU145 prostate cancer cells to Fas(APO1/CD95)-mediated apoptosis through Bcl-2 and MAPK regulation. Cancer Lett 298(1):88–98
Shin DS, Kim HN, Shin KD et al (2009) Cryptotanshinone inhibits constitutive signal transducer and activator of transcription 3 function through blocking the dimerization in DU145 prostate cancer cells. Cancer Res 69:193–202
Hur JM, Shim JS, Jung HJ, Kwon HJ (2005) Cryptotanshinone but not tanshinone IIA inhibits angiogenesisin vitro. Exp Mol Med 37:133–137
Chou TC (2010) Drug combination studies and their synergy quantification using the Chou-Talalay method. Cancer Res 70:440–446
Tracey KJ, Cerami A (1993) Tumor necrosis factor: an updated review of its biology. Crit Care Med 21:S415–422
Fulda S (2010) Evasion of apoptosis as a cellular stress response in cancer. Int J Cell Biol 2010:370835
Garg AK, Aggarwal BB (2002) Reactive oxygen intermediates in TNF signaling. Mol Immunol 39:509–517
Li X, Moody MR, Engel D et al (2000) Cardiac-specific overexpression of tumor necrosis factor-alpha causes oxidative stress and contractile dysfunction in mouse diaphragm. Circulation 102:1690–1696
Bazzoni F, Beutler B (1996) The tumor necrosis factor ligand and receptor families. N Engl J Med 334:1717–1725
Chen B, Cao S, Zhang Y et al (2009) A novel peptide (GX1) homing to gastric cancer vasculature inhibits angiogenesis and cooperates with TNF alpha in anti-tumor therapy. BMC Cell Biol 10:63
Watanabe N, Niitsu Y, Sone H et al (1986) Therapeutic effect of endogenous tumor necrosis factor on ascites Meth A sarcoma. J Immunopharmacol 8:271–283
Chen W, Luo Y, Liu L et al (2010) Cryptotanshinone inhibits cancer cell proliferation by suppressing Mammalian target of rapamycin-mediated cyclin D1 expression and Rb phosphorylation. Cancer Prev Res (Phila) 3:1015–1025
Ye Y, Xu W, Zhong W (2010) Effects of cryptotanshinone on proliferation and apoptosis of Hela cell line of cervical cancer. Zhongguo Zhong Yao Za Zhi 35:118–121
Park IJ, Kim MJ, Park OJ et al (2010) Cryptotanshinone sensitizes DU145 prostate cancer cells to Fas(APO1/CD95)-mediated apoptosis through Bcl-2 and MAPK regulation. Cancer Lett 298:88–98
Aggarwal BB (2004) Nuclear factor-kappaB: the enemy within. Cancer Cell 6:203–208
Amran D, Sanchez Y, Fernandez C et al (2007) Arsenic trioxide sensitizes promonocytic leukemia cells to TNFalpha-induced apoptosis via p38-MAPK-regulated activation of both receptor-mediated and mitochondrial pathways. Biochim Biophys Acta 1773:1653–1663
Bajbouj K, Poehlmann A, Kuester D et al (2009) Identification of phosphorylated p38 as a novel DAPK-interacting partner during TNFalpha-induced apoptosis in colorectal tumor cells. Am J Pathol 175:557–570
Lo YY, Cruz TF (1995) Involvement of reactive oxygen species in cytokine and growth factor induction of c-fos expression in chondrocytes. J Biol Chem 270:11727–11730
Dolado I, Swat A, Ajenjo N, De Vita G, Cuadrado A, Nebreda AR (2007) p38alpha MAP kinase as a sensor of reactive oxygen species in tumorigenesis. Cancer Cell 11:191–205
Acknowledgments
The present study was supported by the Korea Science and Engineering Foundation (KOSEF) grant funded by the Korea government (MEST) (No. 2011-0063466). We thank to Dr. Aggarwal (The University of Texas M.D. Anderson Cancer Center) for providing KBM-5 cell line.
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Ji-Hyun Kim, Soo-** Jeong contributed equally to this work.
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Kim, JH., Jeong, SJ., Kwon, TR. et al. Cryptotanshinone enhances TNF-α-induced apoptosis in chronic myeloid leukemia KBM-5 cells. Apoptosis 16, 696–707 (2011). https://doi.org/10.1007/s10495-011-0605-1
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DOI: https://doi.org/10.1007/s10495-011-0605-1