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Selective induction of apoptosis in various cancer cells irrespective of drug sensitivity through a copper chelate, copper N-(2 hydroxy acetophenone) glycinate: crucial involvement of glutathione

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Abstract

Drug induced toxicity and drug resistance are the major impediments to successful application of cancer chemotherapy. Therefore, selective targeting of the key biochemical events of the malignant cells may have a great therapeutic potential in specifically kill the cancer cells. We have evaluated in vitro the cytotoxic efficacy of a previously reported copper complex viz. copper N-(2-hydroxy acetophenone) glycinate (CuNG) on different drug sensitive and resistant cancer cell lines by MTT, annexin V positivity and caspase 3 activation assays. We have also investigated the underlying signalling events in CuNG mediated apoptosis of cancer cells by Western blotting technique. We have found that CuNG preferentially induces apoptosis to malignant cells irrespective of drug sensitivity and spares the normal cells. Our studies disclose that CuNG causes cellular redox imbalance in cancer cells through depletion of intracellular GSH level. CuNG mediated depletion of intracellular GSH level induces mitochondrial superoxide generation, which detaches cyto C from mitochondrial membrane through lipid peroxidation. The detached cyto C then release into the extra mitochondrial milieu in Bax mediated pathway where CuNG facilitates the binding of Bax through dissociation of hexokinase II from mitochondrial membrane. The present study opens the possibility of developing effective chemotherapeutic drugs by synthesizing numerous chemical compounds capable of targeting cellular redox environment and thus specifically kills cancer cells of broad spectrum.

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Abbreviations

CuNG:

Copper N-(2-hydroxy acetophenone) glycinate

GSH:

Glutathione

Cytochrome C:

Cyto C

HK II:

Hexokinase II

References

  • Armstrong JS, Steinauer KK, Hornung B, Irish JM, Lecane P, Birrell GW et al (2002) Role of glutathione depletion and reactive oxygen species generation in apoptotic signaling in a human B lymphoma cell line. Cell Death Differ 9:252–263

    Article  CAS  PubMed  Google Scholar 

  • Arora KK, Pedersen PL (1988) Functional significance of mitochondrial bound hexokinase in tumor cell metabolism. Evidence for preferential phosphorylation of glucose by intra mitochondrially generated ATP. J Biol Chem 263:17422–17428

    CAS  PubMed  Google Scholar 

  • Arrick BA, Nathan CF (1984) Glutathione metabolism as a determinant of therapeutic efficacy: a review. Cancer Res 44:4224–4232

    CAS  PubMed  Google Scholar 

  • Arrigo AP (1999) Gene expression and the thiol redox state. Free Radic Biol Med 27:936–944

    Article  CAS  PubMed  Google Scholar 

  • Ashkenazi A, Dixit VM (1998) Death receptors: signalling and modulation. Science 281:1305–1308

    Article  CAS  PubMed  Google Scholar 

  • Asumendi A, Morales MC, Alvarez1 A, Aréchaga J, Pérez-Yarza G (2002) Implication of mitochondria-derived ROS and cardiolipin peroxidation in N-(4-hydroxyphenyl) retinamide-induced apoptosis. Br J Cancer 86:1951–1956

    Google Scholar 

  • Basu S, Majumder S, Chatterjee S, Ganguly A, Efferth T, Choudhuri SK (2009) Detection and characterization of a glutathione conjugate of a novel copper complex. In vivo 23:401–4082

    CAS  PubMed  Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

    Article  CAS  PubMed  Google Scholar 

  • Canada A, Herman L, Kidd K, Robertson C, Trump D (1993) Glutathione depletion increases the cytotoxicity of melphalan to PC-3 an androgen-insensitive prostate cancer cell line. Cancer Chemother Pharmaco 32:73–77

    Article  CAS  Google Scholar 

  • Canesi L, Ciacci C, Piccoli G, Stocchi V, Viarengo A, Gallo G (1998) In vitro and in vivo effects of heavy metals on mussel digestive gland hexokinase activity: the role of glutathione. Comp Biochem Physiol C: Pharmacol Toxicol Endocrinol 120:261–268

    Article  CAS  Google Scholar 

  • Chatterjee S, Mookerjee A, Mookerjee Basu J, Chakraborty P, Ganguly A, Adhikary A et al (2009) A novel copper chelate modulates tumor associated macrophages to promote anti-tumor response of T cells. PLoS ONE 4:e7048

    Article  PubMed  Google Scholar 

  • Chen Z, Zhang H, Lu W, Huang P (2009) Role of mitochondria-associated hexokinase II in cancer cell death induced by 3-bromopyruvate. Biochim Biophys Acta 1787:553–560

    Article  CAS  PubMed  Google Scholar 

  • Choudhuri SK (2005) Indian patent entitled ‘‘non toxic anti cancer drug resistance modifying agent and an anticancer kit including the same’.’ Classification no A61 K 31/00. Bigpatents India 1:1–2

    Google Scholar 

  • Drímal J, Zúrová-Nedelcevová J, Knezl V, Sotníková R, Navarová J (2006) Cardiovascular toxicity of the first line cancer chemotherapeutic agents: doxorubicin, cyclophosphamide, streptozotocin and bevacizumab. Neuro Endocrinol Lett 27:176–179

    PubMed  Google Scholar 

  • Efferth T, Sauerbrey A, Olbrich A, Gebhart E, Rauch P, Weber HO et al (2003) Molecular modes of action of artesunate in tumor cell lines. Mol Pharmacol 64:382–394

    Article  CAS  PubMed  Google Scholar 

  • Elo H (1987) Reaction of the antiproliferative and antineoplastic agent trans bis(salicylaldoximato)copper(II) and related chelates with glutathione and cysteine. Correlation between reactivity and biological activity. Inorg Chim Acta 136:L33–L35

    Article  CAS  Google Scholar 

  • Fine RL, Chabner BA (1986) Multidrug resistance. In: Pinedo HM, Chabner BA (eds) Cancer chemotherapy, vol 8. Elsevier, Amsterdam, pp 117–128

    Google Scholar 

  • Ganguly A, Basu S, Chakraborty P, Chatterjee S, Sarkar A, Chatterjee M, Choudhuri SK (2010) Targeting mitochondrial cell death pathway to overcome drug resistance with a newly developed iron chelate. PLoS ONE 5:e11253

    Article  PubMed  Google Scholar 

  • Ganguly A, Basu S, Banerjee K, Chakraborty P, Sarkar A, Chatterjee M, Chaudhuri SK (2011) Redox active copper chelate overcomes multidrug resistance in T-lymphoblastic leukemia cell by triggering apoptosis. Mol BioSyst 7:1701–1712

    Article  CAS  PubMed  Google Scholar 

  • Gillet JP, Efferth T, Steinbach D, Hamels J, de Longueville F, Bertholet V, Remacle J (2004) Microarray-based detection of multidrug resistance in human tumor cells by expression profiling of ATP-binding cassette transporter genes. Cancer Res 64:8987–8993

    Article  CAS  PubMed  Google Scholar 

  • Gurtoo HL, Hipkens JH, Sharma CD (1981) Role of glutathione in the metabolism dependent toxicity and chemotherapy of cyclophosphamide. Cancer Res 41:3584–3591

    CAS  PubMed  Google Scholar 

  • Hamilton TC, Winkler C, Louie K, Batist G, Behrens BC, Tsuruo T et al (1985) Augmentation of adriamycin, melphalan and cisplatin cytotoxicity in drug-resistant and sensitive human ovarian carcinoma cell lines by buthionine sulfoximine mediated glutathione depletion. Biochem Pharmacol 34:2583–2586

    Article  CAS  PubMed  Google Scholar 

  • Hammond CL, Lee TK, Ballatori N (2001) Novel roles for glutathione in gene expression cell death, and membrane transport of organic solutes. J Hepatol 34:946–954

    Article  CAS  PubMed  Google Scholar 

  • Huang ZZ, Chen C, Zeng Z, Yang H, Oh J, Chen L, Lu SC (2001) Mechanism and significance of increased glutathione level in human hepatocellular carcinoma and liver regeneration. FASEB J 15:19–21

    PubMed  Google Scholar 

  • Hutter D, Greene JJ (2000) Influence of the cellular redox state on NF kappa B-regulated gene expression. J Cell Physiol 183:45–52

    Article  CAS  PubMed  Google Scholar 

  • Kuwana T, Mackey MR, Perkins G, Ellisman MH, Latterich M, Schneiter R et al (2002) Bid, Bax, and lipids cooperate to form supramolecular openings in the outer mitochondrial membrane. Cell 111:331–342

    Article  CAS  PubMed  Google Scholar 

  • Latchoumycandane C, Seah QM, Tan RCH, Sattabongkot J, Beerheide W, Boelsterli UA (2006) Leflunomide or A77 1726 protect from acetaminophen-induced cell injury through inhibition of JNK-mediated mitochondrial permeability transition in immortalized human hepatocytes. Toxicol Appl Pharmacol 217:125–133

    Article  CAS  PubMed  Google Scholar 

  • Liu X, Kim CN, Yang J, Jemmerson R, Wang X (1996) Induction of apoptotic program in cell-free extracts: requirement for dATP and cytochrome-c. Cell 86:147–157

    Article  CAS  PubMed  Google Scholar 

  • Majumder S, Panda GS, Choudhuri SK (2003) Synthesis, characterization and biological properties of a novel copper complex. European J Med Chem 38:893–898

    Article  CAS  Google Scholar 

  • Majumder S, Dutta P, Mookerjee A, Choudhuri SK (2006) The role of a novel copper complex in overcoming doxorubicin resistance in Ehrlich ascites carcinoma cells in vivo. Chem Biol Interact 159:90–103

    Article  CAS  PubMed  Google Scholar 

  • Meister A (1983) Selective modification of glutathione metabolism. Science 220:472–477

    Article  CAS  PubMed  Google Scholar 

  • Mookerjee A, Basu JM, Majumder S, Chatterjee S, Panda GS, Dutta P et al (2006) A novel copper complex induces ROS generation in doxorubicin resistant Ehrlich ascitis carcinoma cells and increases activity of antioxidant enzymes in vital organs in vivo. BMC Cancer 6:267–277

    Article  PubMed  Google Scholar 

  • Müller M, Meijer C, Zaman GJ, Borst P, Scheper RJ, Mulder NH et al (1994) Overexpression of the gene encoding the multidrug resistance-associated protein results in increased ATP-dependent glutathione S-conjugate transport. Proc Natl Acad Sci U S A 91:13033–13037

    Article  PubMed  Google Scholar 

  • Nakashima RA, Mangan PS, Colombini M, Pedersen PL (1986) Hexokinase receptor complex in hepatoma mitochondria: evidence from N,N′-dicyclohexylcarbodiimide-labeling studies for the involvement of the pore-forming protein VDAC. Biochemistry 25:1015–1021

    Article  CAS  PubMed  Google Scholar 

  • Nechushtan A, Smith CL, Hsu YT, Youle RJ (1999) Conformation of the Bax C-terminus regulates subcellular location and cell death. EMBO J 18:2330–2341

    Article  CAS  PubMed  Google Scholar 

  • Nomura K, Imai H, Koumura T, Kobayashi T, Nakagawa Y (2000) Mitochondrial phospholipid hydroperoxide glutathione peroxidase inhibits the release of cytochrome c from mitochondria by suppressing the peroxidation of cardiolipin in hypoglycaemia-induced apoptosis. Biochem J 351:183–193

    Article  CAS  PubMed  Google Scholar 

  • Ott M, Robertson JD, Gogvadze V, Zhivotovsky B, Orrenius S (2002) Cytochrome c release from mitochondria proceeds by a two-step process. Proc Natl Acad Sci USA 99:1259–1263

    Article  CAS  PubMed  Google Scholar 

  • Ozols RF, O’Dwyer PJ, Hamilton TC, Young RC (1990) The role of glutathione in drug resistance. Cancer Treat Rev 17:45–50

    Article  CAS  PubMed  Google Scholar 

  • Pastorino JG, Shulga N, Hoek JB (2002) Mitochondrial binding of hexokinase II inhibits Bax-induced cytochrome C release and apoptosis. J Biol Chem 277:7610–7618

    Article  CAS  PubMed  Google Scholar 

  • Rae TD, Schmidt PJ, Pufahl RA, Culotta VC, O’Halloran TV (1999) Undetectable intracellular free copper: the requirement of a copper chaperone for superoxide dismutase. Science 284:805–808

    Article  CAS  PubMed  Google Scholar 

  • Rhodes T, Twentyman PR (1992) A study of ethacrynic acid as a potential modifier of melphalan and cisplatin sensitivity in human lung cancer parental and drug-resistant cell lines. Br J Cancer 65:684–690

    Article  CAS  PubMed  Google Scholar 

  • Sargent DJ, Niedzwiecki D, O’Connell MJ, Schilsky RL (2001) Recommendation for caution with irinotecan, fluorouracil, and leucovorin for colorectal cancer. N Engl J Med 345:144–145

    Article  CAS  PubMed  Google Scholar 

  • Schneider E, Yamazaki H, Sinha BK, Cowan KH (1995) Buthionine sulphoximine-mediated sensitisation of etoposide-resistant human breast cancer MCF7 cells overexpressing the multidrug resistance-associated protein involves increased drug accumulation. Br J Cancer 71:738–743

    Article  CAS  PubMed  Google Scholar 

  • Schreck R, Rieberl P, Baeuerle PA (1991) Reactive oxygen intermediates as apparently widely used messengers in the activation of the NF-κB transcription factor and HIV-1. EMBO J 10:2247–2258

    CAS  PubMed  Google Scholar 

  • Suzukake K, Vistica BP, Vistica DT (1983) Dechlorination of l-phenylalanine mustard by sensitive and resistant tumor cells and its relationship to intracellular glutathione content. Biochem Pharmacol 32:165–167

    Article  CAS  PubMed  Google Scholar 

  • Szatrowski TP, Nathan CF (1991) Production of large amounts of hydrogen peroxide by human tumor cells. Cancer Res 51:794–798

    CAS  PubMed  Google Scholar 

  • Trachootham D, Zhou Y, Zhang H, Demizu Y, Chen Z, Pelicano H et al (2006) Selective killing of oncogenically transformed cells through a ROS mediated mechanism by β-phenylethyl isothiocyanate. Cancer Cell 10:421–452

    Article  Google Scholar 

  • Wang W, Ballatori N (1998) Endogenous glutathione conjugates: occurrence and biological functions. Pharmacol Rev 50:335–356

    CAS  PubMed  Google Scholar 

  • Wang CY, Mayo MW, Korneluk RG, Goeddel DV, Baldwin AS Jr (1998) NF-kappaB anti apoptosis: induction of TRAF1 and TRAF2 and c-IAP1 and c-IAP2 to suppress caspase-8 activation. Science 281:1680–1683

    Article  CAS  PubMed  Google Scholar 

  • Wright SC, Wang H, Wei QS, Kinder DH, Larrick JW (1998) Bcl-2-mediated resistance to apoptosis is associated with glutathione induced inhibition of AP24 activation of nuclear DNA fragmentation. Cancer Res 58:5570–5576

    CAS  PubMed  Google Scholar 

  • Wu HY, Kang YJ (1998) Inhibition of buthionine sulfoximine-enhanced doxorubicin toxicity in metallothionein overexpressing transgenic mouse heart. J Pharmacol Exp Ther 287:515–520

    CAS  PubMed  Google Scholar 

  • Wu MX, Ao Z, Prasad KV, Wu R, Schlossman SF (1998) IEX-1L, an apoptosis inhibitor involved in NF-kappaB-mediated cell survival. Science 281:998–1001

    Article  CAS  PubMed  Google Scholar 

  • Xu L, Koumenis IL, Tilly JL, Giffard RG (1999) Overexpression of bcl-xL protects astrocytes from glucose deprivation and is associated with higher glutathione, ferritin, and iron levels. Anesthesiology 91:1036–1046

    Article  CAS  PubMed  Google Scholar 

  • Yamamoto K, Masubuchi Y, Narimatsu S, Kobayashi S, Horie T (2002) Toxicity of ethacrynic acid in isolated rat hepatocytes. Toxicol In Vitro 16:151–158

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

Authors acknowledge the help of Prof. Marian Valko, Slovak Technical University, Bratislava, Slovakia for doing EPR work. This investigation received financial support from Indian Council of Medical Research (ICMR), New Delhi, No. 5/13/18/2007/NCD-III, 3/2/2/177/2009-NCDIII and 5/13/53/2008/NCDIII.

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Department of In Vitro Carcinogenesis and Cellular Chemotherapy, Chittaranjan National Cancer Institute, S.P. Mukherjee Road, Kolkata, 700026, India

    Shilpak Chatterjee, Paramita Chakraborty, Kaushik Banerjee, Abhinaba Sinha & Soumitra Kumar Choudhuri

  2. Department of Molecular Medicine, Bose Institute, Kolkata, India

    Arghya Adhikary & Tanya Das

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  1. Shilpak Chatterjee
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  2. Paramita Chakraborty
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  3. Kaushik Banerjee
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  5. Arghya Adhikary
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  6. Tanya Das
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  7. Soumitra Kumar Choudhuri
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Correspondence to Soumitra Kumar Choudhuri.

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Chatterjee, S., Chakraborty, P., Banerjee, K. et al. Selective induction of apoptosis in various cancer cells irrespective of drug sensitivity through a copper chelate, copper N-(2 hydroxy acetophenone) glycinate: crucial involvement of glutathione. Biometals 26, 517–534 (2013). https://doi.org/10.1007/s10534-013-9637-z

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