Skip to main content

Advertisement

SpringerLink
Log in

Identification of the molecular mechanisms underlying the cytotoxic action of a potent platinum metallointercalator

  • Original Article
  • Published:
Journal of Chemical Biology

Abstract

Platinum-based DNA metallointercalators are structurally different from the covalent DNA binders such as cisplatin and its derivatives but have potent in vitro activity in cancer cell lines. However, limited understanding of their molecular mechanisms of cytotoxic action greatly hinders their further development as anticancer agents. In this study, a lead platinum-based metallointercalator, [(5,6-dimethyl-1,10-phenanthroline) (1S,2S-diaminocyclohexane)platinum(II)]2+ (56MESS) was found to be 163-fold more active than cisplatin in a cisplatin-resistant cancer cell line. By using transcriptomics in a eukaryotic model organism, yeast Saccharomyces cerevisiae, we identified 93 genes that changed their expressions significantly upon exposure of 56MESS in comparison to untreated controls (p ≤ 0.05). Bioinformatic analysis of these genes demonstrated that iron and copper metabolism, sulfur-containing amino acids and stress response were involved in the cytotoxicity of 56MESS. Follow-up experiments showed that the iron and copper concentrations were much lower in 56MESS-treated cells compared to controls as measured by inductively coupled plasma optical emission spectrometry. Deletion mutants of the key genes in the iron and copper metabolism pathway and glutathione synthesis were sensitive to 56MESS. Taken together, the study demonstrated that the cytotoxic action of 56MESS is mediated by its ability to disrupt iron and copper metabolism, suppress the biosynthesis of sulfur-containing amino acids and attenuate cellular defence capacity. As these mechanisms are in clear contrast to the DNA binding mechanism for cisplatin and its derivative, 56MESS may be able to overcome cisplatin-resistant cancers. These findings have provided basis to further develop the platinum-based metallointercalators as anticancer agents.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  1. Alic N, Felder T, Temple MD, Gloeckner C, Higgins VJ, Briza P, Dawes IW (2004) Genomewide transcriptional responses to a lipid hydroperoxide: adaptation occurs without induction of oxidant defenses. Free Radic Biol Med 37:23–35

    Article  CAS  Google Scholar 

  2. Ang WH, Khalaila I, Allardyce CS, Juillerat-Jeanneret L, Dyson PJ (2005) Rational design of platinum(IV) compounds to overcome glutathione-S-transferase mediated drug resistance. J Am Chem Soc 127:1382–1383

    Article  CAS  Google Scholar 

  3. Askwith C, Kaplan J (1998) Iron and copper transport in yeast and its relevance to human disease. Trends Biochem Sci 23:135–138

    Article  CAS  Google Scholar 

  4. Balendiran G, Dabur R, Fraser D (2004) The role of glutathione in cancer. Cell Biochem and Funct 22:343–352

    Article  CAS  Google Scholar 

  5. Barabas K, Milner R, Lurie D, Adin C (2008) Cisplatin: a review of toxicities and therapeutic applications. Vet Comp Oncol 6:1–18

    Article  CAS  Google Scholar 

  6. Basu A, Krishnamurthy S (2010) Cellular responses to cisplatin-induced DNA damage. J Nucleic Acids. doi:10.4061/2010/201367

  7. Boyer J, Allen WL, McLean EG, Wilson PM, McCulla A, Moore S, Longley DB, Caldas C, Johnston PG (2006) Pharmacogenomic identification of novel determinants of response to chemotherapy in colon cancer. Cancer Res 66:2765–2777

    Article  CAS  Google Scholar 

  8. Brodie CR, Grant Collins J, Aldrich-Wright JR (2004) DNA binding and biological activity of some platinum(II) intercalating compounds containing methyl-substituted 1,10-phenanthrolines. Dalton Trans:1145–1152

  9. Caba E, Dickinson DA, Warnes GR, Aubrecht J (2005) Differentiating mechanisms of toxicity using global gene expression analysis in Saccharomyces cerevisiae. Mutat Res 575:34–46

    Article  CAS  Google Scholar 

  10. Calsou P, Barret J-M, Cros S, Salles B (1993) DNA excision–repair synthesis is enhanced in a murine leukemia L1210 cell line resistant to cisplatin. Eur J Biochem 211:403–409

    Article  CAS  Google Scholar 

  11. Charizanis C, Juhnke H, Krems B, Entian K (1999) The mitochondrial cytochrome c peroxidase Ccp1 of Saccharomyces cerevisiae is involved in conveying an oxidative stress signal to the transcription factor Pos9 (Skn7). Mol Gen Genet 262:437–447

    Article  CAS  Google Scholar 

  12. De Virgilio C, Bürckert N, Bell W, Jenö P, Boller TAW (1993) Disruption of TPS2, the gene encoding the 100-kDa subunit of the trehalose-6-phosphate synthase/phosphatase complex in Saccharomyces cerevisiae, causes accumulation of trehalose-6-phosphate and loss of trehalose-6-phosphate phosphatase activity. Eur J Biochem 212:315–323

    Article  Google Scholar 

  13. Dillon CT (2011) Synchrontron radiation x-ray spectroscopy for investigations of introcellular metallointercalators: x-ray fluorescence imaging and x-ray absorption spectroscopy. In: Aldrich-Wright JR (ed) Metallointercalators, synthesis and techniques to probe their interactions with biomolecules. Springer, New York, pp 273–298

    Google Scholar 

  14. Djinovic K, Gatti G, Coda A, Antolini L, Pelosi G, Desideri A, Falconi M, Marmocchi F, Rotilio GMB (1992) Crystal structure of yeast Cu, Zn superoxide dismutase. Crystallographic refinement at 2.5 A resolution. J Mol Biol 225:791–809

    Article  CAS  Google Scholar 

  15. Eastman A (1999) The mechanism of action of cisplatin: from adducts to apoptosis. In: Lippert B (ed) Cisplatin: chemistry and biochemistry of a leading anticancer drug. VHCA, Zürich, pp 111–134

    Google Scholar 

  16. Einhorn LH (2002) Curing metastatic testicular cancer. Proc Natl Acad Sci USA 99:4592–4595

    Article  CAS  Google Scholar 

  17. Farquharson MJ, Al-Ebraheem A, Falkenberg G, Leek R, Harris AL, Bradley DA (2008) The distribution of trace elements Ca, Fe, Cu and Zn and the determination of copper oxidation state in breast tumour tissue using muSRXRF and muXANES. Phys Med Biol 53:3023–3037

    Article  CAS  Google Scholar 

  18. Fisher DM, Bednarski PJ, Grünert R, Turner P, Fenton RR, Aldrich-Wright JR (2007) Chiral platinum(II) metallointercalators with potent in vitro cytotoxic activity. ChemMedChem 2:488–495

    Article  CAS  Google Scholar 

  19. Fisher DM, Fenton RR, Aldrich-Wright JR (2008) In vivo studies of a platinum(II) metallointercalator. Chem Commun 43:5613–5615

    Article  Google Scholar 

  20. Freshney R (2005) Culture of animal cells: a manual of basic technique, 5th edn. Wiley-Liss, New York, pp 359–373

    Book  Google Scholar 

  21. Fu D, Beeler TJ, Dunn TM (1995) Sequence, mapping and disruption of CCC2, a gene that cross complements the Ca2 + 517 -sensitive phenotype of csg1 mutants and encodes a P-type ATPase belonging to the Cu2 + 518 -ATPase subfamily. Yeast 11:283–292

    Article  CAS  Google Scholar 

  22. Gasch A, Spellman P, Kao C, Carmel-Harel O, Eisen M, Storz G, Botstein D, Brown P (2000) Genomic expression programs in the response of yeast cells to environmental changes. Mol Biol Cell 11:4241–4257

    CAS  Google Scholar 

  23. Gasch AP, Huang M, Metzner S, Botstein D, Elledge SJ, Brown PO (2001) Genomic expression responses to DNA-damaging agents and the regulatory role of the yeast ATR homolog Mec1p. Mol Biol Cell 12:2987–3003

    CAS  Google Scholar 

  24. Georgatsou E, Alexandraki D (1994) Two distinctly regulated genes are required for ferric reduction, the first step of iron uptake in Saccharomyces cerevisiae. Mol Cell Biol 14:3065–3073

    CAS  Google Scholar 

  25. Gibson G, Muse SV (2009) A prime of genome science, 3rd edn. Sinauer, Sunderland, pp 191–258

    Google Scholar 

  26. Godwin AK, Meister A, O'dwyer PJ, Huang CS, Hamilton TC, Anderson ME (1992) High resistance to cisplatin in human ovarian cancer cell lines is associated with marked increase of glutathione synthesis. Proc Natl Acad Sci USA 89:3070–3074

    Article  CAS  Google Scholar 

  27. Grant CM (2001) Role of the glutathione/glutaredoxin and thioredoxin systems in yeast growth and response to stress conditions. Mol Microbiol 39:533–541

    Article  CAS  Google Scholar 

  28. Hansen J, Johannesen PF (2000) Cysteine is essential for transcriptional regulation of the sulfur assimilation genes in Saccharomyces cerevisiae. Mol Gen Genet 263:535–542

    Article  CAS  Google Scholar 

  29. Head JF, Wang F, Elliott RL (1997) Antineoplastic drugs that interfere with iron metabolism in cancer cells. Adv Enzyme Regul 37:147–169

    Article  CAS  Google Scholar 

  30. Heymann P, Ernst JF, Winkelmann G (2000) Identification and substrate specificity of a ferrichrome-type siderophore transporter (Arn1p) in Saccharomyces cerevisiae. FEMS Microbiol Lett 186:221–227

    Article  CAS  Google Scholar 

  31. Ho PS, Hoffman BM, Solomon N, Kang CH, Margoliash E (1984) Kinetics and energetics of intramolecular electron transfer in yeast cytochrome c peroxidase. Biochemistry 23:4122–4128

    Article  CAS  Google Scholar 

  32. Hu GF (1998) Copper stimulates proliferation of human endothelial cells under culture. J Cell Biochem 69:326–335

    Article  CAS  Google Scholar 

  33. Ionescu JG, Novotny J, Stejskal V, Latsch A, Blaurock-Busch E, Eisenmann-Klein M (2006) Increased levels of transition metals in breast cancer tissue. Neuroendocrin Lett 21:36–39

    Google Scholar 

  34. Ishida S, McCormick F, Smith-McCune K, Hanahan D (2010) Enhancing tumor-specific uptake of the anticancer drug cisplatin with a copper chelator. Cancer Cell 17:574–583

    Article  CAS  Google Scholar 

  35. Jaramillo D, Buck DP, Collins JG, Fenton RR, Stootman FH, Wheate NJ, Aldrich-Wright JR (2006) Synthesis, characterisation and biological activity of chiral platinum(II) complexes. Eur J Inorg Chem 2006:839–849

    Article  Google Scholar 

  36. Jauniaux JC, Urrestarazu LA, Wiame JM (1978) Arginine metabolism in Saccharomyces cerevisiae: subcellular localization of the enzymes. J Bacteriol 133:1096–1107

    CAS  Google Scholar 

  37. Johnson SW, Ferry KV, Hamilton TC (1998) Recent insights into platinum drug resistance in cancer. Drug Resist Update 1:243–254

    Article  CAS  Google Scholar 

  38. Jones EW, Fink GR (1982) Regulation of amino acid and nucleotide biosynthesis in yeast. In: Strathern JN, Jones EW, Broach JR (eds) The molecular biology of the yeast Saccharomyces: metabolism and gene expression. CSHL, New York, pp 181–299

    Google Scholar 

  39. Kabat GC, Rohan T (2007) Does excess iron play a role in breast carcinogenesis? An unresolved hypothesis. Cancer Cause Control 18:1047–1053

    Article  Google Scholar 

  40. Kelland LR (2007) The resurgence of platinum-based cancer chemotherapy. Nat Rev Cancer 7:573–584

    Article  CAS  Google Scholar 

  41. Kemp JD (1997) Iron deprivation and cancer: a view beginning with studies of monoclonal antibodies against the transferrin receptor. Histol Histopathol 12:291–296

    CAS  Google Scholar 

  42. Kemp S, Wheate NJ, Buck DP, Nikac M, Collins JG, Aldrich-Wright JR (2007) The effect of ancillary ligand chirality and phenanthroline functional group substitution on the cytotoxicity of platinum(II)-based metallointercalators. J Inorg Biochem 101:1049–1058

    Article  CAS  Google Scholar 

  43. Kemp S, Wheate NJ, Wang S, Collins J, Ralph S, Day A, Higgins VJ, Aldrich-Wright JR (2007) Encapsulation of platinum(II)-based DNA intercalators within cucurbit[6,7,8]urils. J Biol Inorg Chem 12:969–979

    Article  CAS  Google Scholar 

  44. Kicic A, Chua ACG, Baker E (2001) Effect of iron chelators on proliferation and iron uptake in hepatoma cells. Cancer 92:3093–3110

    Article  CAS  Google Scholar 

  45. Krause-Heuer AM, Grünert R, Kühne S, Buczkowska M, Wheate NJ, Le Pevelen DD, Boag LR, Fisher DM, Kasparkova J, Malina J, Bednarski PJ, Brabec V, Aldrich-Wright JR (2009) Studies of the mechanism of action of platinum(II) complexes with potent cytotoxicity in human cancer cells. J Med Chem 52:5474–5484

    Article  CAS  Google Scholar 

  46. Kuras L, Barbey R, Thomas D (1997) Assembly of a bZIP–bHLH transcription activation complex: formation of the yeast Cbf1–Met4–Met28 complex is regulated through Met28 stimulation of Cbf1 DNA binding. EMBO J 16:2441–2451

    Article  CAS  Google Scholar 

  47. Le NT, Richardson DR (2002) The role of iron in cell cycle progression and the proliferation of neoplastic cells. Biochim Biophys Acta 1603:31–46

    CAS  Google Scholar 

  48. Lee J, Straffon MJ, Jang T, Higgins VJ, Grant CM, Dawes IW (2001) The essential 582 and ancillary role of glutathione in Saccharomyces cerevisiae analysed using a grande gsh1 disruptant strain. FEMS Yeast Res 1:57–65

    CAS  Google Scholar 

  49. Lesuisse E, Blaiseau PL, Dancis A, Camadro J (2001) Siderophore uptake and use by the yeast Saccharomyces cerevisiae. Microbiology 147:289–298

    CAS  Google Scholar 

  50. Lim AL, Powers-Lee SG (1996) Requirement for the carboxyl-terminal domain of Saccharomyces cerevisiae carbamoyl-phosphate synthetase. J Biol Chem 271:11400–11409

    Article  CAS  Google Scholar 

  51. Mai B, Breeden L (1997) Xbp1, a stress-induced transcriptional repressor of the Saccharomyces cerevisiae Swi4/Mbp1 family. Mol Cell Biol 17:6491–6501

    CAS  Google Scholar 

  52. Margiotta N, Ostuni R, Gandin V, Marzano C, Piccinonna S, Natile G (2009) Synthesis, characterization, and cytotoxicity of dinuclear platinum-bisphosphonate complexes to be used as prodrugs in the local treatment of bone tumours. Dalton Trans 48:10904–10913

    Article  Google Scholar 

  53. McFadyen WD, Wakelin LPG, Roos IAG, Leopold VA (1985) Activity of platinum(II) intercalating agents against murine leukemia L1210. J Med Chem 28:1113–1116

    Article  CAS  Google Scholar 

  54. Meister A, Anderson ME (1983) Glutathione. Annu Rev Biochem 52:711–760

    Article  CAS  Google Scholar 

  55. Moretto J, Chauffert B, Ghiringhelli F, Aldrich-Wright JR, Bouyer F (2010) Discrepancy between in vitro and in vivo antitumor effect of a new platinum(II) metallointercalator. Invest New Drugs 29:1164–1176

    Article  Google Scholar 

  56. Odani T, Shimma Y, Wang XHYJ (1997) Mannosylphosphate transfer to cell wall mannan is regulated by the transcriptional level of the MNN4 gene in Saccharomyces cerevisiae. FEBS Lett 420:186–190

    Article  CAS  Google Scholar 

  57. Protchenko O, Philpott C (2003) Regulation of intracellular heme levels by HMX1, a homologue of heme oxygenase, in Saccharomyces cerevisiae. J Biol Chem 278:36582–36587

    Article  CAS  Google Scholar 

  58. Rabik CA, Dolan ME (2007) Molecular mechanisms of resistance and toxicity associated with platinating agents. Cancer Treat Rev 33:9–23

    Article  CAS  Google Scholar 

  59. Reizenstein P (1991) Iron, free radicals and cancer: a review. Med Oncol Tumor Pharmacother 8:229–233

    CAS  Google Scholar 

  60. Richardson J, Thomas KA, Rubin BH, Richardson DC (1975) Crystal structure of bovine Cu, Zn superoxide dismutase at 3 A resolution: chain tracing and metal ligands. Proc Nat Acad Sci 72:1349–1353

    Article  CAS  Google Scholar 

  61. Richon VM, Schulte N, Eastman A (1987) Multiple mechanisms of resistance to cis diamminedichloroplatinum(II) in murine leukemia L1210 Cells. Cancer Res 47:2056–2061

    CAS  Google Scholar 

  62. Robbins E, Pederson T (1970) Iron: its intracellular localization and possible role in cell division. Proc Natl Acad Sci USA 66:1244–1251

    Article  CAS  Google Scholar 

  63. Rodriguez-Manzaneque MT, Ros J, Cabiscol E, Sorribas A, Herrero E (1999) Grx5 glutaredoxin plays a central role in protection against protein oxidative damage in Saccharomyces cerevisiae. Mol Cell Biol 19:8180–8190

    CAS  Google Scholar 

  64. Rutherford JC, Jaron S, Winge DR (2003) Aft1p and Aft2p mediate iron-responsive gene expression in yeast through related promoter elements. J Biol Chem 278:27636–27643

    Article  CAS  Google Scholar 

  65. Seligman P, Schleicher R, Siriwardana G, Domenico J, Gelfand E (1993) Effects of agents that inhibit cellular iron incorporation on bladder cancer cell proliferation. Blood 82:1608–1617

    CAS  Google Scholar 

  66. Siddik Z (2003) Cisplatin: mode of cytotoxic action and molecular basis of 623 resistance. Oncogene 22:7265–7279

    Article  CAS  Google Scholar 

  67. Sies H (1999) Glutathione and its role in cellular functions. Free Radic Biol Med 27:916–921

    Article  CAS  Google Scholar 

  68. Simon J, Bedalov A (2004) Yeast as a model system for anticancer drug discovery. Nat Rev Cancer 4:1–8

    Article  Google Scholar 

  69. Stearman R, Yuan DS, Yamaguchi-Iwai Y, Klausner RD, Dancis A (1996) A permease–oxidase complex involved in high-affinity iron uptake in yeast. Science 271:1552–1557

    Article  CAS  Google Scholar 

  70. Teixeira M, Monteiro P, Jain P, Tenreiro S, Fernandes A, Mira N, Alenquer M, Freitas A, Oliveira A, Sa-Correia I (2006) The YEASTRACT database: a tool for the analysis of transcription regulatory associations in Saccharomyces cerevisiae. Nucleic Acids Res 34:D446–451

    Article  CAS  Google Scholar 

  71. Thomas-Chollier M, Sand O, Turatsinze J, Janky R, Defrance M, Vervisch E, Brohe’e S, van Helden J (2008) RSAT: regulatory sequence analysis tools. Nucleic Acids Res 36:W119–W127

    Article  CAS  Google Scholar 

  72. Toyokuni S (1996) Iron-induced carcinogenesis: the role of redox regulation. Free Radic Bio Med 20:553–566

    Article  CAS  Google Scholar 

  73. van Helden J (2003) Regulatory sequence analysis tools. Nucleic Acids Res 31:3593–3596

    Article  Google Scholar 

  74. Van Ho A, Ward DM, Kaplan J (2002) Transition metal transport in yeast. Annu Rev Microbiol 56:237–261

    Article  Google Scholar 

  75. van Loon AP, Pesold-Hunt B, Schatz G (1986) A yeast mutant lacking mitochondrial manganese superoxide dismutase is hypersensitive to oxygen. Proc Natl Acad Sci USA 83:3820–3824

    Article  Google Scholar 

  76. Wang D, Lippard SJ (2005) Cellular processing of platinum anticancer drugs. Nat Rev Drug Discov 4:308–320

    Article  Google Scholar 

  77. Weinberg ED (1999) Iron therapy and cancer. Kidney Int 55:S131–S134

    Article  Google Scholar 

  78. Wheate NJ, Collins JG (2005) Multi-nuclear platinum drugs: a new paradigm in chemotherapy. Curr Med Chem - Anti-Cancer Agents 5:267–279

    Article  CAS  Google Scholar 

  79. Wheate NJ, Taleb RI, Krause-Heuer AM, Cook RL, Wang S, Higgins VJ, Aldrich-Wright JR (2007) Novel platinum(II)-based anticancer complexes and molecular hosts as their drug delivery vehicles. Dalton Trans 43:5055–5064

    Article  Google Scholar 

  80. Yamaguchi-Iwai Y, Ueta R, Fukunaka A, Sasaki R (2002) Subcellular localization Aft1 transcription factor responds to iron status in Saccharomyces cerevisiae. J Biol Chem 277:18914–18918

    Article  CAS  Google Scholar 

Download references

Acknowledgements

The authors thank the Australian Government for the International Science Linkage Grant and the University of Western Sydney for financial support through internal research grants (JRAW and MW). S.W. was supported by APA Award and UWS Top-Up Award from the University of Western Sydney. We also thank Ramaciotti Centre at UNSW for Affymetrix microarray, Dr. C. Cullinane from the Peter MacCallum Cancer Centre for supplying the L1210cisR cancer cell line and K. McNamara for technical assistance.

Author information

Authors and Affiliations

  1. School of Biomedical and Health Science, University of Western Sydney, Locked Bag 1797, Sydney, Penrith, NSW 2751, Australia

    Shaoyu Wang, Vincent J. Higgins, Janice R. Aldrich-Wright & Ming J. Wu

  2. School of Medicine, University of Western Sydney, Locked Bag 1797, Sydney, Penrith, NSW 2751, Australia

    Shaoyu Wang

  3. Ramaciotti Centre for Gene Function Analysis, School of Biotechnology and Biomolecular Sciences, University of New South Wales, Paddington, NSW 2052, Australia

    Vincent J. Higgins

Authors
  1. Shaoyu Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Vincent J. Higgins
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Janice R. Aldrich-Wright
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ming J. Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Janice R. Aldrich-Wright or Ming J. Wu.

Electronic Supplementary Materials

Below is the link to the electronic supplementary material.

ESM 1

(PDF 30 kb)

ESM 2

(PDF 126 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Wang, S., Higgins, V.J., Aldrich-Wright, J.R. et al. Identification of the molecular mechanisms underlying the cytotoxic action of a potent platinum metallointercalator. J Chem Biol 5, 51–61 (2012). https://doi.org/10.1007/s12154-011-0070-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12154-011-0070-x

Keywords

Search

Navigation