Abstract
3,5-Dimethyl-7H-furo[3,2-g]chromen-7-one (DMFC) is a coumarin derivative with anti-cancer activity against human hepatoma cells, but the mechanisms underlying DMFC function in cancer suppression is unknown. In this study, we aimed at elucidating the molecular mechanisms underlying DMFC anti-cancer activity and determining whether DMFC is effective in suppression of drug-resistant human hepatocellular carcinoma. We show here that DMFC effectively suppresses both the parent and the multidrug-resistant hepatoma cell growth in vitro and DMFC suppresses hepatoma cell growth at least in part through inducing tumor cell apoptosis. In the molecular level, we observed that DMFC treatment decreases Bcl-2 level by a post-transcriptional mechanism and activates Bim transcription to increase Bim mRNA and protein level in hepatoma cells. Furthermore, co-immunoprecipitation studies revealed that DMFC-induced Bim interrupts interactions between Bcl-2 and Bax and between Mcl-1 and Bak, resulting in dissociation of Bax from Bcl-2 and Bak from Mcl-1 and subsequent activation of both Bax and Bak. Activation of Bax and Bak leads to mitochondrial outer membrane permeabilization and cytochrome c release. Consistent with its potent apoptosis-inducing activity, DMFC exhibited potent activity against the multidrug-resistant hepatoma xenograft growth in vivo. Therefore, we determine that DMFC suppresses hepatoma growth through decreasing Bcl-2 and increasing Bim to induce tumor cell apoptosis and hold great promise for further development as a therapeutic agent to treat chemoresistant hepatoma.
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References
El-Serag HB (2011) Hepatocellular carcinoma. N Engl J Med 365:1118–1127
Lise M, Pilati P, Da PP, Mocellin S, Nitti D, Corazzino S (2003) Treatment options for liver metastases from colorectal cancer. J Exp Clin Cancer Res 22:149–156
van den Hoven AF, Lam MG, Jernigan S, van den Bosch MA, Buckner GD (2015) Innovation in catheter design for intra-arterial liver cancer treatments results in favorable particle–fluid dynamics. J Exp Clin Cancer Res 34:74
Wang CK, Lin YF, Tai CJ et al (2015) Integrated treatment of aqueous extract of Solanum nigrum-potentiated cisplatin-and doxorubicin-induced cytotoxicity in human hepatocellular carcinoma cells. Evid Based Complement Alternat Med 2015:675270
Wang Z, Li J, Ji Y, An P, Zhang S, Li Z (2013) Traditional herbal medicine: a review of potential of inhibitory hepatocellular carcinoma in basic research and clinical trial. Evid Based Complement Alternat Med 2013:268963–268969
Kerr JF, Wyllie AH, Currie AR (1972) Apoptosis: a basic biological phenomenon with wide-ranging implications in tissue kinetics. Br J Cancer 26:239–257
Danial NN (2007) BCL-2 family proteins: critical checkpoints of apoptotic cell death. Clin Cancer Res 13:7254–7263
Jin Z, El-Deiry WS (2005) Overview of cell death signaling pathways. Cancer Biol Ther 4:139–163
Cory S, Adams JM (2002) The Bcl2 family: regulators of the cellular life-or-death switch. Nat Rev Cancer 2:647–656
Elmore S (2007) Apoptosis: a review of programmed cell death. Toxicol Pathol 35:495–516
Wang K, Yin XM, Chao DT, Milliman CL, Korsmeyer SJ (1996) BID: a novel BH3 domain-only death agonist. Genes Dev 10:2859–2869
Danial NN, Korsmeyer SJ (2004) Cell death: critical control points. Cell 116:205–219
Liu X, Dai S, Zhu Y, Marrack P, Kappler JW (2003) The structure of a Bcl-xL/Bim fragment complex: implications for Bim function. Immunity 19:341–352
Petros AM, Nettesheim DG, Wang Y et al (2000) Rationale for Bcl-xL/Bad peptide complex formation from structure, mutagenesis, and biophysical studies. Protein Sci 9:2528–2534
Reed JC, Zha H, Aime-Sempe C, Takayama S, Wang HG (1996) Structure–function analysis of Bcl-2 family proteins. Regulators of programmed cell death. Adv Exp Med Biol 406:99–112
Hamacher-Brady A, Brady NR (2015) Bax/Bak-dependent, Drp1-independent targeting of XIAP into inner-mitochondrial compartments counteracts Smac-dependent effector caspase activation. J Biol Chem 290:22005–22018
Zerp SF, Stoter TR, Hoebers FJ et al (2015) Targeting anti-apoptotic Bcl-2 by AT-101 to increase radiation efficacy: data from in vitro and clinical pharmacokinetic studies in head and neck cancer. Radiat Oncol 10:158
Li X, Zeng X, Sun J et al (2014) Imperatorin induces Mcl-1 degradation to cooperatively trigger Bax translocation and Bak activation to suppress drug-resistant human hepatoma. Cancer Lett 348:146–155
Sun JG, Chen CY, Luo KW et al (2011) 3,5-Dimethyl-H-furo[3,2-g]chromen-7-one as a potential anticancer drug by inducing p53-dependent apoptosis in human hepatoma HepG2 cells. Chemotherapy 57:162–172
Cuvillier O, Rosenthal DS, Smulson ME, Spiegel S (1998) Sphingosine 1-phosphate inhibits activation of caspases that cleave poly(ADP-ribose) polymerase and lamins during Fas- and ceramide-mediated apoptosis in Jurkat T lymphocytes. J Biol Chem 273:2910–2916
Reed JC (2006) Proapoptotic multidomain Bcl-2/Bax-family proteins: mechanisms, physiological roles, and therapeutic opportunities. Cell Death Differ 13:1378–1386
Lucken-Ardjomande S, Martinou JC (2005) Newcomers in the process of mitochondrial permeabilization. J Cell Sci 118:473–483
Chen S, Dai Y, Harada H, Dent P, Grant S (2007) Mcl-1 down-regulation potentiates ABT-737 lethality by cooperatively inducing Bak activation and Bax translocation. Cancer Res 67:782–791
Cuconati A, Mukherjee C, Perez D, White E (2003) DNA damage response and MCL-1 destruction initiate apoptosis in adenovirus-infected cells. Genes Dev 17:2922–2932
Antonsson B, Conti F, Ciavatta A et al (1997) Inhibition of Bax channel-forming activity by Bcl-2. Science 277:370–372
Chen L, Willis SN, Wei A et al (2005) Differential targeting of prosurvival Bcl-2 proteins by their BH3-only ligands allows complementary apoptotic function. Mol Cell 17:393–403
van Delft MF, Wei AH, Mason KD et al (2006) The BH3 mimetic ABT-737 targets selective Bcl-2 proteins and efficiently induces apoptosis via Bak/Bax if Mcl-1 is neutralized. Cancer Cell 10:389–399
Cai J, Yang J, Jones DP (1998) Mitochondrial control of apoptosis: the role of cytochrome c. Biochim Biophys Acta 1366:139–149
Itchaki G, Brown JR (2016) The potential of venetoclax (ABT-199) in chronic lymphocytic leukemia. Ther Adv Hematol 7(5):270–287
Khaw S, Merino D, Anderson M, Glaser S, Bouillet P, Roberts A et al (2014) Both leukaemic and normal peripheral B lymphoid cells are highly sensitive to the selective pharmacological inhibition of prosurvival Bcl-2 with ABT-199. Leukemia 28:1207–1215
Vogler M, Butterworth M, Majid A, Walewska R, Sun X, Dyer M et al (2009) Concurrent up-regulation of BCL-XL and BCL2A1 induces approximately 1000-fold resistance to ABT-737 in chronic lymphocytic leukemia. Blood 113:4403–4413
Vogler M, Dinsdale D, Dyer M, Cohen G (2013) ABT-199 selectively inhibits BCL2 but not BCL2L1 and efficiently induces apoptosis of chronic lymphocytic leukaemic cells but not platelets. Br J Haematol 163:139–142
Congreve M, Carr R, Murray C, Jhoti H (2003) A ‘rule of three’ for fragment-based lead discovery. Drug Discov Today 8(19):876–877
Sarosiek KA, Chi X, Bachman JA et al (2013) BID preferentially activates BAK while BIM preferentially activates BAX, affecting chemotherapy response. Mol Cell 51:751–765
Acknowledgements
This study is supported by National Natural Science Foundation of China (No. 31570811) Natural Science Foundation of Zhejiang Province (LY15C020001) and Si-Yuan Foundation.
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All applicable international, national, and institutional guidelines for the care and use of animals were followed. This article does not contain any studies with human participants performed by any of the authors.
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Jun Xiang, Zhe Wang and Qianqia Liu have contributed equally to this work.
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Xiang, J., Wang, Z., Liu, Q. et al. DMFC (3,5-dimethyl-7H-furo[3,2-g]chromen-7-one) regulates Bim to trigger Bax and Bak activation to suppress drug-resistant human hepatoma. Apoptosis 22, 381–392 (2017). https://doi.org/10.1007/s10495-016-1331-5
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DOI: https://doi.org/10.1007/s10495-016-1331-5