Esculetin Induces Apoptosis in Human Leukemia Kasumi-1 cells through Caspase 3 Activation
DOI:
https://doi.org/10.56511/JIPBS.2015.v2.i03.pp273-289Abstract
Objective: AML alone accounts for 20% cases of acute leukemia in children and 80% cases in adults. Among many genetic alterations, the most frequently reported translocation is between chromosomes 8 and 21, t(8; 21) or AML1-ETO. In the past decade, the anti–proliferative effect of various natural compounds including esculetin (a coumarin), has been reported in several leukemic cell lines. We have recently shown that esculetin reduces the half-life of AML1-ETO chimeric mRNA as well as mutated c-Kit transcripts in human monocytic leukemia kasumi-1 cell line harbouring the t(8;21) translocation. In the present study we established the antiproliferating activity of esculetin on kasumi-1 cell line. Methods: In this study, cytotoxicity of the esculetin on kausmi-1 cells was investigated. The ability of esculetin to induce cell cycle arrest, alter mitochondrial potential, activate Caspase cascade and to express apoptotic markers was investigated through western blotting analysis and flow cytometry methods.Results and conclusion: The half maximal inhibitory concentration of esculetin in kasumi-1 cells was found to be 100 μM. Ecsuletin arrested the cell cycle at G0/G1 phase in kasumi-1 cells. A significant increase in mitochondrial membrane potential and cytosolic release of cytochrome C was observed in ecsuletin treated kasumi-1 cells. This was accompanied with activation of Caspase 3 and Caspase 8 and enhanced cleavage of phospholipase C (PLC)γ-1. Annexin V apoptotic assay further corrobrated our results that esculetin mediated cytoxicity which is accompanied by cleavage of Caspase 3 is due to apoptosis in kausmi-1 cells.
Keywords:
Esculetin, AML1-ETO, Kasumi-1, Apoptosis, Caspase 3, Caspase 8Downloads
References
Peng XM, Damu GL, Zhou C: Current developments of coumarin compounds in medicinal chemistry. Curr Pharm Des 2013, 19(21):3884-3930.
Chang WS, Lin CC, Chuang SC, Chiang HC: Superoxide anion scavenging effect of coumarins. Am J Chin Med 1996, 24(1):1117.
JM Y, J X, Y Z, HD S, ZW L: Chemical components from Ceratostigma willmottianum. Journal of Natural Products 1997, 60(10):1031-1033.
Kemal C, Louis-Flamberg P, KrupinskiOlsen R, Shorter AL: Reductive inactivation of soybean lipoxygenase 1 by catechols: a possible mechanism for regulation of lipoxygenase activity. Biochemistry 1987, 26(22):7064-7072.
Shin E, Choi KM, Yoo HS, Lee CK, Hwang BY, Lee MK: Inhibitory effects of coumarins from the stem barks of Fraxinus rhynchophylla on adipocyte differentiation in 3T3-L1 cells. Biol Pharm Bull 2010, 33(9):1610-1614.
Yang JY, Della-Fera MA, Baile CA: Esculetin induces mitochondria-mediated apoptosis in 3T3-L1 adipocytes. Apoptosis 2006, 11(8):1371-1378.
Kim SH, Kang KA, Zhang R, Piao MJ, Ko DO, Wang ZH, Chae SW, Kang SS, Lee KH, Kang HK et al : Protective effect of esculetin against oxidative stress-induced cell damage via scavenging reactive oxygen species. Acta Pharmacol Sin 2008, 29(11):1319-1326.
Venugopala KN, Rashmi V, Odhav B: Review on natural coumarin lead compounds for their pharmacological activity. Biomed Res Int 2013, 2013:963248.
Park SS, Park SK, Lim JH, Choi YH, Kim WJ, Moon SK: Esculetin inhibits cell proliferation through the Ras/ERK1/2 pathway in human colon cancer cells. Oncol Rep 2011, 25(1):223-230.
Anand JR, Rijhwani H, Malapati K, Kumar P, Saikia K, Lakhar M: Anticancer activity of esculetin via-modulation of Bcl-2 and NF- ?B expression in benzo[a]pyrene induced lung carcinogenesis in mice. . Biomedicine & Preventive Nutrition 2013, 3(2):107-102.
Kok SH, Yeh CC, Chen ML, Kuo MY: Esculetin enhances TRAIL-induced apoptosis through DR5 upregulation in human oral cancer SAS cells. Oral Oncol 2009, 45(12):1067-1072.
Al-Jorany KHA, Saeed NA-HAAH: Cytotoxic effect of esculetin on myeloma and myelogenous leukemic cells. Iraqi Journal of Cancer and Medical Genetics 2012, 5(2):147152.
Chu CY, Tsai YY, Wang CJ, Lin WL, Tseng TH: Induction of apoptosis by esculetin in human leukemia cells. Eur J Pharmacol 2001, 416(1-2):25-32.
Wang CJ, Hsieh YJ, Chu CY, Lin YL, Tseng TH: Inhibition of cell cycle progression in human leukemia HL-60 cells by esculetin. Cancer Lett 2002, 183(2):163-168.
Park C, Jin CY, Kim GY, Choi IW, Kwon TK, Choi BT, Lee SJ, Lee WH, Choi YH: Induction of apoptosis by esculetin in human leukemia U937 cells through activation of JNK and ERK. Toxicol Appl Pharmacol 2008, 227(2):219-228.
Lee SH, Park C, Jin CY, Kim GY, Moon SK, Hyun JW, Lee WH, Choi BT, Kwon TK, Yoo YH et al : Involvement of extracellular signal-related kinase signaling in esculetin induced G1 arrest of human leukemia U937 cells. Biomed Pharmacother 2008, 62(10):723-729.
Park C, Jin CY, Kwon HJ, Hwang HJ, Kim GY, Choi IW, Kwon TK, Kim BW, Kim WJ, Choi YH: Induction of apoptosis by esculetin in human leukemia U937 cells: roles of Bcl-2 and extracellular-regulated kinase signaling. Toxicol In Vitro 2010, 24(2):486494.
Park C, Hyun S-K, Shin W-J, Chung K-T, Choi B-T, Kwon H-J, Hwang H-J, Kim B-W, Park D-I, Lee W-H et al : Esculetin Induces Apoptosis through Caspase-3 Activation in Human Leukemia U937 Cells. Journal of Life Science 2009, 19(2):249-255.
Miyoshi H, Shimizu K, Kozu T, Maseki N, Kaneko Y, Ohki M: t(8;21) breakpoints on chromosome 21 in acute myeloid leukemia are clustered within a limited region of a single gene, AML1. Proc Natl Acad Sci U S A 1991, 88(23):10431-10434.
Erickson P, Gao J, Chang KS, Look T, Whisenant E, Raimondi S, Lasher R, Trujillo J, Rowley J, Drabkin H: Identification of breakpoints in t(8;21) acute myelogenous leukemia and isolation of a fusion transcript, AML1/ETO, with similarity to Drosophila segmentation gene, runt. Blood 1992, 80(7):1825-1831.
Nisson PE, Watkins PC, Sacchi N: Transcriptionally active chimeric gene derived from the fusion of the AML1 gene and a novel gene on chromosome 8 in t(8;21) leukemic cells. Cancer Genet Cytogenet 1992, 63(2):81-88.
Miyoshi H, Kozu T, Shimizu K, Enomoto K, Maseki N, Kaneko Y, Kamada N, Ohki M: The t(8;21) translocation in acute myeloid leukemia results in production of an AML1MTG8 fusion transcript. Embo j 1993, 12(7):2715-2721.
Klampfer L, Zhang J, Zelenetz AO, Uchida H, Nimer SD: The AML1/ETO fusion protein activates transcription of BCL-2. Proc Natl Acad Sci U S A 1996, 93(24):14059-14064.
Westendorf JJ, Yamamoto CM, Lenny N, Downing JR, Selsted ME, Hiebert SW: The t(8;21) fusion product, AML-1-ETO, associates with C/EBP-alpha, inhibits C/EBP-alpha-dependent transcription, and blocks granulocytic differentiation. Mol Cell Biol 1998, 18(1):322-333.
Sinenko SA, Hung T, Moroz T, Tran QM, Sidhu S, Cheney MD, Speck NA, Banerjee U: Genetic manipulation of AML1-ETOinduced expansion of hematopoietic precursors in a Drosophila model. Blood 2010, 116(22):4612-4620.
Li X, Xu YB, Wang Q, Lu Y, Zheng Y, Wang YC, Lubbert M, Zhao KW, Chen GQ: Leukemogenic AML1-ETO fusion protein upregulates expression of connexin 43: the role in AML 1-ETO-induced growth arrest in leukemic cells. J Cell Physiol 2006, 208(3):594-601.
Elsasser A, Franzen M, Kohlmann A, Weisser M, Schnittger S, Schoch C, Reddy VA, Burel S, Zhang DE, Ueffing M et al : The fusion protein AML1-ETO in acute myeloid leukemia with translocation t(8;21) induces c-jun protein expression via the proximal AP-1 site of the c-jun promoter in an indirect, JNK-dependent manner. Oncogene 2003, 22(36):5646-5657.
Dohner H, Estey EH, Amadori S, Appelbaum FR, Buchner T, Burnett AK, Dombret H, Fenaux P, Grimwade D, Larson RA et al : Diagnosis and management of acute myeloid leukemia in adults: recommendations from an international expert panel, on behalf of the European LeukemiaNet. Blood 2010, 115(3):453-474.
Fey MF, Greil R, Jost LM: ESMO Minimum Clinical Recommendations for the diagnosis, treatment and follow-up of acute myeloblastic leukemia (AML) in adult patients. Ann Oncol 2005, 16 Suppl 1:i4849.
Kolitz JE: Current therapeutic strategies for acute myeloid leukaemia. Br J Haematol 2006, 134(6):555-572.
Sawney S, Arora R, Aggarwal KK, Saluja D: Esculetin Downregulates the Expression of AML1-ETO and C-Kit in Kasumi-1 Cell Line by Decreasing Half-Life of mRNA. Journal of Oncology 2015, 2015:8.
Holden P, Horton WA: Crude subcellular fractionation of cultured mammalian cell lines. BMC research notes 2009, 2:243.
Vieira L, Oliveira V, Ambrosio AP, Marques B, Pereira AM, Hagemeijer A, Boavida MG: Translocation (8;17;15;21)(q22;q23;q15;q22) in acute myeloid leukemia (M2). a four-way variant of t(8;21). Cancer Genet Cytogenet 2001, 128(2):104-107.
Rubnitz JE, Raimondi SC, Halbert AR, Tong X, Srivastava DK, Razzouk BI, Pui CH, Downing JR, Ribeiro RC, Behm FG: Characteristics and outcome of t(8;21)- positive childhood acute myeloid leukemia: a single institution's experience. Leukemia 2002, 16(10):2072-2077.
Reikvam H, Hatfield KJ, Kittang AO, Hovland R, Bruserud O: Acute myeloid leukemia with the t(8;21) translocation: clinical consequences and biological implications. J Biomed Biotechnol 2011, 2011:104631.
Specchia G, Albano F, Anelli L, Zagaria A, Liso A, La Starza R, Mancini M, Sebastio L, Giugliano E, Saglio G et al : Insertions generating the 5'RUNX1/3'CBFA2T1 gene in acute myeloid leukemia cases show variable breakpoints. Genes Chromosomes Cancer 2004, 41(1):86-91.
Elagib KE, Goldfarb AN: Oncogenic pathways of AML1-ETO in acute myeloid leukemia: multifaceted manipulation of marrow maturation. Cancer Lett 2007, 251(2):179-186.
Sun D, Urrabaz R, Buzello C, Nguyen M: Effects of cisplatin on expression of DNA ligases in MiaPaCa human pancreatic cancer cells. Biochem Biophys Res Commun 2002, 298(4):537-544.
Yoshikawa R, Kusunoki M, Yanagi H, Noda M, Furuyama JI, Yamamura T, HashimotoTamaoki T: Dual antitumor effects of 5fluorouracil on the cell cycle in colorectal carcinoma cells: a novel target mechanism concept for pharmacokinetic modulating chemotherapy. Cancer Res 2001, 61(3):1029-1037.
Gallagher BM, Jr.: Microtubule-stabilizing natural products as promising cancer therapeutics. Curr Med Chem 2007, 14(28):2959-2967.
Singh RP, Agarwal R: Natural flavonoids targeting deregulated cell cycle progression in cancer cells. Curr Drug Targets 2006, 7(3):345-354.
Rasul A, Khan M, Yu B, Ma T, Yang H: Xanthoxyletin, a coumarin induces S phase arrest and apoptosis in human gastric adenocarcinoma SGC-7901 cells. Asian Pac J Cancer Prev 2011, 12(5):1219-1223.
Murphy AN, Bredesen DE, Cortopassi G, Wang E, Fiskum G: Bcl-2 potentiates the maximal calcium uptake capacity of neural cell mitochondria. Proc Natl Acad Sci U S A 1996, 93(18):9893-9898.
Indran IR, Tufo G, Pervaiz S, Brenner C: Recent advances in apoptosis, mitochondria and drug resistance in cancer cells. Biochim Biophys Acta 2011, 1807(6):735-745.
Park JW, Choi YJ, Suh SI, Baek WK, Suh MH, Jin IN, Min DS, Woo JH, Chang JS, Passaniti A et al : Bcl-2 overexpression attenuates resveratrol-induced apoptosis in U937 cells by inhibition of caspase-3 activity. Carcinogenesis 2001, 22(10):1633-1639.
Lawen A: Apoptosis-an introduction. Bioessays 2003, 25(9):888-896.
Vermes I, Haanen C, Steffens-Nakken H, Reutelingsperger C: A novel assay for apoptosis. Flow cytometric detection of phosphatidylserine expression on early apoptotic cells using fluorescein labelled Annexin V. J Immunol Methods 1995, 184(1):39-51.
Aubry JP, Blaecke A, Lecoanet-Henchoz S, Jeannin P, Herbault N, Caron G, Moine V, Bonnefoy JY: Annexin V used for measuring apoptosis in the early events of cellular cytotoxicity. Cytometry 1999, 37(3):197204.
Tao D, Wu J, Feng Y, Qin J, Hu J, Gong J: New method for the analysis of cell cyclespecific apoptosis. Cytometry A 2004, 57(2):70-74.
Zhou GB, Kang H, Wang L, Gao L, Liu P, Xie J, Zhang FX, Weng XQ, Shen ZX, Chen J et al : Oridonin, a diterpenoid extracted from medicinal herbs, targets AML1-ETO fusion protein and shows potent antitumor activity with low adverse effects on t(8;21) leukemia in vitro and in vivo. Blood 2007, 109(8):3441-3450.
Corsello SM, Roti G, Ross KN, Chow KT, Galinsky I, DeAngelo DJ, Stone RM, Kung AL, Golub TR, Stegmaier K: Identification of AML1-ETO modulators by chemical genomics. Blood 2009, 113(24):6193-6205.
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