Establishment of an Arsenic Trioxide—Resistant Human Leukemia Cell Line That Shows Multidrug Resistance
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We have established an arsenic trioxide (As2O3)-resistant cell line (K562/AS-3) derived from the human leukemia cell line K562. K562/AS-3 was sequentially cultured with increasing concentrations of As2O3 up to 3.5 µM and then cloned by the limiting dilution method. K562/AS-3 was found to be about 7-fold more resistant to As2O3 than the parent cells (IC50 = 12.9 µM for K562/AS-3 and 1.8 µM for K562), and also showed cross resistance to VP-16 and vincristine. The multidrug resistance—associated protein (MRP1) gene was found to be overexpressed, but the MDR gene was not detected. MRP1 function was evaluated by measuring calcein acetoxymethyl ester (calcein-AM) efflux, and by verifying its inhibition by MK571, a potent MRP inhibitor. In addition, an increase of the total intracellular glutathione content was found in K562/AS-3. The resistance of K562/AS-3 to As2O3 was reversed by the addition of MK571, but not by verapamil. K562/AS-3 may be useful for studying the mechanism of the anticancer effect of As2O3 and how to overcome As2O3-resistance.
- Seo T, Fukushima T, Inoue H, et al. Long-term follow-up of the clinical efficacy of chemotherapy for acute myeloid leukemia at a single institute. J Infect Chemother. 2001;7:156–162. CrossRef
- Soignet SL, Maslak P, Wang ZG, et al. Complete remission after treatment of acute promyelocytic leukemia with arsenic trioxide. N Engl J Med. 1998;339:1341–1348. CrossRef
- Shen ZX, Chen GQ, Ni JH, et al. Use of arsenic trioxide (As2O3) in the treatment of acute promyelocytic leukemia (APL): II. Clinical efficacy and pharmacokinetics in relapsed patients. Blood. 1997;89:3354–3360.
- Niu C, Yan H, Yu T, et al. Studies on treatment of acute promyelocytic leukemia with arsenic trioxide: remission induction, follow- up, and molecular monitoring in 11 newly diagnosed and 47 relapsed acute promyelocytic leukemia patients. Blood. 1999;94:3315–3324.
- Chen GQ, Zhu J, Shi XG, et al. In vitro studies on cellular and molecular mechanisms of arsenic trioxide (As2O3) in the treatment of acute promyelocytic leukemia: As2O3 induces NB4 cell apoptosis with downregulation of Bcl-2 expression and modulation of PML-RAR alpha/PML proteins. Blood. 1996;88:1052–1061.
- Shao W, Fanelli M, Ferrara FF, et al. Arsenic trioxide as an inducer of apoptosis and loss of PML/RAR alpha protein in acute promyelocytic leukemia cells. J Natl Cancer Inst. 1998;90:124–133. CrossRef
- Zhu J, Koken MH, Quignon F, et al.Arsenic-induced PML targeting onto nuclear bodies: implications for the treatment of acute promyelocytic leukemia. Proc Natl Acad Sci U S A. 1997;94:3978–3983. CrossRef
- Seo T, Urasaki Y, Takemura H, Ueda T. Arsenic trioxide circumvents multidrug resistance based on different mechanisms in human leukemia cell lines. Anticancer Res. 2005;25:991–998.
- Zhang Y, Nie L. Studies of apoptosis of malignant lymphoma cells induced by arsenic trioxide. Cell Biol Int. 2001;25:1003–1006. CrossRef
- Zhang TC, Cao EH, Li JF, Ma W, Qin JF. Induction of apoptosis and inhibition of human gastric cancer MGC-803 cell growth by arsenic trioxide. Eur J Cancer. 1999;35:1258–1263. CrossRef
- Akao Y, Nakagawa Y, Akiyama K. Arsenic trioxide induces apoptosis in neuroblastoma cell lines through the activation of caspase 3 in vitro. FEBS Lett. 1999;455:59–62. CrossRef
- Ravandi F, van Besien K. Clinical activity of arsenic trioxide in Burkitt-like lymphoma. Leukemia. 2003;17:271–272. CrossRef
- Bahlis NJ, McCafferty-Grad J, Jordan-McMurry I, et al. Feasibility and correlates of arsenic trioxide combined with ascorbic acid- mediated depletion of intracellular glutathione for the treatment of relapsed/refractory multiple myeloma. Clin Cancer Res. 2002;8:3658–3668.
- Oketani M, Kohara K, Tuvdendorj D, et al. Inhibition by arsenic trioxide of human hepatoma cell growth. Cancer Lett. 2002;183:147–153. CrossRef
- Brambila EM, Achanzar WE, Qu W, Webber MM, Waalkes MP. Chronic arsenic-exposed human prostate epithelial cells exhibit stable arsenic tolerance: mechanistic implications of altered cellular glutathione and glutathione S-transferase. Toxicol Appl Pharmacol. 2002;183:99–107. CrossRef
- Kitamura K, Minami Y, Yamamoto K, et al. Involvement of CD95- independent caspase 8 activation in arsenic trioxide-induced apoptosis. Leukemia. 2000;14:1743–1750. CrossRef
- Lee TC, Wei ML, Chang WJ, et al. Elevation of glutathione levels and glutathione S-transferase activity in arsenic-resistant Chinese hamster ovary cells. In Vitro Cell Dev Biol. 1989;25:442–448. CrossRef
- Flens MJ, Izquierdo MA, Scheffer GL, et al. Immunochemical detection of the multidrug resistance-associated protein MRP in human multidrug-resistant tumor cells by monoclonal antibodies. Cancer Res. 1994;54:4557–4563.
- Feller N, Kuiper CM, Lankelma J, et al. Functional detection of MDR1/P170 and MRP/P190-mediated multidrug resistance in tumour cells by flow cytometry. Br J Cancer. 1995;72:543–549.
- Zhou DC, Zittoun R, Marie JP. Expression of multidrug resistance- associated protein (MRP) and multidrug resistance (MDR1) genes in acute myeloid leukemia. Leukemia. 1995;9:1661–1666.
- Salerno M, Petroutsa M, Garnier-Suillerot A. The MRP1-mediated effluxes of arsenic and antimony do not require arsenic-glutathione and antimony-glutathione complex formation. J Bioenerg Biomembr. 2002;34:135–145. CrossRef
- Vernhet L, Allain N, Payen L, Anger JP, Guillouzo A, Fardel O. Resistance of human multidrug resistance-associated protein 1-overexpressing lung tumor cells to the anticancer drug arsenic trioxide. Biochem Pharmacol. 2001;61:1387–1391. CrossRef
- Takeshita A, Shinjo K, Naito K, et al. P-glycoprotein (P-gp) and multidrug resistance-associated protein 1 (MRP1) are induced by arsenic trioxide (As(2)O(3)), but are not the main mechanism of As(2)O(3)-resistance in acute promyelocytic leukemia cells. Leukemia. 2003;17:648–650. CrossRef
- Lorico A, Bertola A, Baum C, Fodstad O, Rappa G. Role of the Multidrug Resistance Protein 1 in protection from heavy metal oxyanions: investigations in vitro and in MRP1-deficient mice. Biochem Biophys Res Commun. 2002;291:617–622. CrossRef
- Dai J, Weinberg RS, Waxman S, Jing Y. Malignant cells can be sensitized to undergo growth inhibition and apoptosis by arsenic trioxide through modulation of the glutathione redox system. Blood. 1999;93:268–277.
- Jing Y, Dai J, Chalmers-Redman RM, Tatton WG, Waxman S. Arsenic trioxide selectively induces acute promyelocytic leukemia cell apoptosis via a hydrogen peroxide-dependent pathway. Blood. 1999;94:2102–2111.
- Scott N, Hatlelid KM, MacKenzie NE, Carter DE. Reactions of arsenic(III) and arsenic(V) species with glutathione. Chem Res Toxicol. 1993;6:102–106. CrossRef
- Yang CH, Kuo ML, Chen JC, Chen YC.Arsenic trioxide sensitivity is associated with low level of glutathione in cancer cells. Br J Cancer. 1999;81:796–799. CrossRef
- Nakagawa Y, Akao Y, Morikawa H, et al. Arsenic trioxide-induced apoptosis through oxidative stress in cells of colon cancer cell lines. Life Sci. 2002;70:2253–2269. CrossRef
- Davison K, Cote S, Mader S, Miller WH. Glutathione depletion overcomes resistance to arsenic trioxide in arsenic-resistant cell lines. Leukemia. 2003;17:931–940. CrossRef
- Kala SV, Neely MW, Kala G, et al. The MRP2/cMOAT transporter and arsenic-glutathione complex formation are required for biliary excretion of arsenic. J Biol Chem. 2000;275:33404–33408. CrossRef
- Establishment of an Arsenic Trioxide—Resistant Human Leukemia Cell Line That Shows Multidrug Resistance
International Journal of Hematology
Volume 85, Issue 1 , pp 26-31
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- Arsenic trioxide
- Arsenic resistance
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- Author Affiliations
- 1. First Department of Internal Medicine, University of Fukui, Fukui, Japan
- 2. Division of Transfusion Medicine, Faculty of Medical Sciences, University of Fukui, 23-3 Matuokashimoaizuki, Eiheiji-cho, Yoshida-gun, 910-1193, Fukui, Japan