pp 1–11 | Cite as

Cellular responses of BRCA1-defective HCC1937 breast cancer cells induced by the antimetastasis ruthenium(II) arene compound RAPTA-T

  • Tidarat Nhukeaw
  • Khwanjira Hongthong
  • Paul J. Dyson
  • Adisorn RatanaphanEmail author


An organometallic ruthenium(II) arene compound, Ru(η6-toluene)(PTA)Cl2 (PTA = 1,3,5-triaza-7-phosphaadamantane), termed RAPTA-T, exerts promising antimetastatic properties. In this study, the effects of RAPTA-T on BRCA1-defective HCC1937 breast cancer cells have been investigated, and compared to its effects on BRCA1-competent MCF-7 breast cancer cells. RAPTA-T showed a very low cytotoxicity against both tested cells. Ruthenium is found mostly in the cytoplasmic compartment of both cells. Flow cytometric analysis reveals that the compound arrests the growth of both cells by triggering the G2/M phase that led to the induction of apoptosis. At equimolar concentrations, RAPTA-T causes much more cellular BRCA1 damage in HCC1937 than in MCF-7 cells, suppressing the expression of BRCA1 mRNA in both cell lines with the subsequent down-regulation of the BRCA1 protein. Interestingly, RAPTA-T exhibits an approximately fivefold greater ability to suppress the expression of the BRCA1 protein in HCC1937 than in MCF-7 cells. These data provide insights into the molecular mechanisms by which RAPTA-T exerts its effects on BRCA1-associated breast cancer cells.


Ruthenium complexes BRCA1 Breast cancer Cell cycle Apoptosis BRCA1 expression 



This research work was supported by the National Research Council of Thailand (PHA610093S and PHA590396S), Prince of Songkla University (PHA6202079S), and the Graduate school, Prince of Songkla University. We would like to thank the Pharmaceutical Laboratory Service Center and Department of Pharmaceutical Chemistry, Faculty of Pharmaceutical Sciences, Prince of Songkla University for providing research facilities.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflicts of interest.

Human and animal rights

The article does not contain any studies with human participants or animals performed by any of the authors.


  1. 1.
    Siegel RL, Miller KD, Jemal A (2016) Cancer statistics: 2016. CA Cancer J Clin 66:7–30. CrossRefGoogle Scholar
  2. 2.
    Li Y, Li S, Chen J et al (2014) Comparative epigenetic analyses reveal distinct patterns of oncogenic pathways activation in breast cancer subtypes. Hum Mol Genet 23:5378–5393. CrossRefGoogle Scholar
  3. 3.
    Perou CM, Sørlie T, Eisen MB et al (2000) Molecular portraits of human breast tumours. Nature 406:747–752. CrossRefGoogle Scholar
  4. 4.
    Severson TM, Peeters J, Majewski I et al (2015) BRCA1-like signature in triple negative breast cancer: molecular and clinical characterization reveals subgroups with therapeutic potential. Mol Oncol 9:1528–1538. CrossRefGoogle Scholar
  5. 5.
    Bhattacharyya A, Ear US, Koller BH, Weichselbaum RR, Bishop DK (2000) The breast cancer susceptibility gene BRCA1 is required for subnuclear assembly of Rad51 and survival following treatment with the DNA cross-linking agent cisplatin. J Biol Chem 275:23899–23903. CrossRefGoogle Scholar
  6. 6.
    Elstrodt F, Hollestelle A, Nagel JH et al (2006) BRCA1 mutation analysis of 41 human breast cancer cell lines reveals three new deleterious mutants. Cancer Res 66:41–45. CrossRefGoogle Scholar
  7. 7.
    Tassone P, Tagliaferri P, Perricelli A et al (2003) BRCA1 expression modulates chemosensitivity of BRCA1-defective HCC1937 human breast cancer cells. Br J Cancer 88:1285–1291. CrossRefGoogle Scholar
  8. 8.
    Domagala P, Huzarski T, Lubinski J, Gugala K, Domagala W (2011) Immunophenotypic predictive profiling of BRCA1-associated breast cancer. Virchows Arch 458:55–64. CrossRefGoogle Scholar
  9. 9.
    Esteller M, Silva JM, Dominguez G et al (2000) Promoter hypermethylation and BRCA1 inactivation in sporadic breast and ovarian tumors. J Natl Cancer Inst 92:564–569. CrossRefGoogle Scholar
  10. 10.
    Stefansson OA, Villanueva A, Vidal A, Martí L, Esteller M (2012) BRCA1 epigenetic inactivation predicts sensitivity to platinum-based chemotherapy in breast and ovarian cancer. Epigenetics 7:1225–1229. CrossRefGoogle Scholar
  11. 11.
    Veeck J, Ropero S, Setien F et al (2010) BRCA1 CpG island hypermethylation predicts sensitivity to poly(adenosine diphosphate)-ribose polymerase inhibitors. J Clin Oncol 28:563–564. CrossRefGoogle Scholar
  12. 12.
    Font A, Taron M, Gago JL et al (2011) BRCA1 mRNA expression and outcome to neoadjuvant cisplatin-based chemotherapy in bladder cancer. Ann Oncol 22:139–144. CrossRefGoogle Scholar
  13. 13.
    Isakoff SJ, Mayer EL, He L et al (2015) TBCRC009: a multicenter phase II clinical trial of platinum monotherapy with biomarker assessment in metastatic triple-negative breast cancer. J Clin Oncol 33:1902–1909. CrossRefGoogle Scholar
  14. 14.
    Carozzi VA, Marmiroli P, Cavaletti G (2010) The role of oxidative stress and anti-oxidant treatment in platinum-induced peripheral neurotoxicity. Curr Cancer Drug Targets 10:670–682. CrossRefGoogle Scholar
  15. 15.
    Tezcan S, Izzettin FV, Sancar M, Yumuk PF, Turhal S (2013) Nephrotoxicity evaluation in outpatients treated with cisplatin-based chemotherapy using a short hydration method. Pharmacol Pharm 4:296–302. CrossRefGoogle Scholar
  16. 16.
    Dhillon KK, Swisher EM, Taniguchi T (2011) Secondary mutations of BRCA1/2 and drug resistance. Cancer Sci 102:663–669. CrossRefGoogle Scholar
  17. 17.
    Swisher EM, Sakai W, Karlan BY, Wurz K, Urban N, Taniguchi T (2008) Secondary BRCA1 mutations in BRCA1-mutated ovarian carcinomas with platinum resistance. Cancer Res 68:2581–2586. CrossRefGoogle Scholar
  18. 18.
    Hongthong K, Ratanaphan A (2016) BRCA1-associated triple-negative breast cancer and potential treatment for ruthenium-based compounds. Curr Cancer Drug Targets 16:606–617. CrossRefGoogle Scholar
  19. 19.
    Bergamo A, Gaiddon C, Schellens JH, Beijnen JH, Sava G (2012) Approaching tumour therapy beyond platinum drugs: status of the art and perspectives of ruthenium drug candidates. J Inorg Biochem 106:90–99. CrossRefGoogle Scholar
  20. 20.
    Leijen S, Burgers SA, Baas P et al (2015) Phase I/II study with ruthenium compound NAMI-A and gemcitabine in patients with non-small cell lung cancer after first line therapy. Invest New Drugs 33:201–214. CrossRefGoogle Scholar
  21. 21.
    Murray BS, Babakb MV, Hartingerb CG, Dyson PJ (2016) The development of RAPTA compounds for the treatment of tumors. Coord Chem Rev 306:86–114. CrossRefGoogle Scholar
  22. 22.
    Scolaro C, Bergamo A, Brescacin L et al (2005) In vitro and in vivo evaluation of ruthenium(II)-arene PTA complexes. J Med Chem 48:4161–4171. CrossRefGoogle Scholar
  23. 23.
    Chatterjee S, Kundu S, Bhattacharyya A, Hartinger CG, Dyson PJ (2008) The ruthenium(II)-arene compound RAPTA-C induces apoptosis in EAC cells through mitochondrial and p53-JNK pathways. J Biol Inorg Chem 13:1149–1155. CrossRefGoogle Scholar
  24. 24.
    Weiss A, Berndsen RH, Dubois M et al (2014) In vivo anti-tumor activity of the organometallic ruthenium(II)-arene complex [Ru(η6-p-cymene)Cl2(pta)] (RAPTA-C) in human ovarian and colorectal carcinomas. Chem Sci 5:4742–4748. CrossRefGoogle Scholar
  25. 25.
    Weiss A, Bonvin D, Berndsen RH et al (2015) Angiostatic treatment prior to chemo-or photodynamic therapy improves anti-tumor efficacy. Sci Rep 5:8990. CrossRefGoogle Scholar
  26. 26.
    Weiss A, Ding X, van Beijnum JR et al (2015) Rapid optimization of drug combinations for the optimal angiostatic treatment of cancer. Angiogenesis 18:233–244. CrossRefGoogle Scholar
  27. 27.
    Adhireksan Z, Davey GE, Campomanes P et al (2014) Ligand substitutions between ruthenium-cymene compounds can control protein versus DNA targeting and anticancer activity. Nat Commun 5:3462–3475. CrossRefGoogle Scholar
  28. 28.
    Wu B, Ong MS, Groessl M et al (2011) A ruthenium antimetastasis agent forms specific histone protein adducts in the nucleosome core. Chemistry 17:3562–3566. CrossRefGoogle Scholar
  29. 29.
    Groessl M, Tsybin YO, Hartinger CG, Keppler BK, Dyson PJ (2010) Ruthenium versus platinum: interactions of anticancer metallodrugs with duplex oligonucleotides characterised by electrospray ionisation mass spectrometry. J Biol Inorg Chem 15:677–688. CrossRefGoogle Scholar
  30. 30.
    Nowak-Sliwinska P, van Beijnum JR, Casini A et al (2011) Organometallic ruthenium(II) arene compounds with antiangiogenic activity. J Med Chem 54:3895–3902. CrossRefGoogle Scholar
  31. 31.
    Bergamo A, Masi A, Dyson PJ, Sava G (2008) Modulation of the metastatic progression of breast cancer with an organometallic ruthenium compound. Int J Oncol 33:1281–1289. Google Scholar
  32. 32.
    Lee RFS, Chernobrovkin A, Rutishauser D et al (2017) Expression proteomics study to determine metallodrug targets and optimal drug combinations. Sci Rep 7:1590. CrossRefGoogle Scholar
  33. 33.
    Ratanaphan A, Nhukeaw T, Hongthong K, Dyson PJ (2017) Differential cytotoxicity, cellular uptake, apoptosis and inhibition of BRCA1 expression of BRCA1-defective and sporadic breast cancer cells induced by an anticancer ruthenium(II)-arene compound, RAPTA-EA1. Anticancer Agents Med Chem 17:212–220. CrossRefGoogle Scholar
  34. 34.
    Koch RB (1969) Fractionation of olfactory tissue homogenates. Isolation of a concentrated plasma membrane fraction. J Neurochem 16:145–157. CrossRefGoogle Scholar
  35. 35.
    Nhukeaw T, Temboot P, Hansongnern K, Ratanaphan A (2014) Cellular responses of BRCA1-defective and triple-negative breast cancer cells and in vitro BRCA1 interactions induced by metallo-intercalator ruthenium(II) complexes containing chloro-substituted phenylazopyridine. BMC Cancer 14:73. CrossRefGoogle Scholar
  36. 36.
    Ratanaphan A, Canyuk B, Wasiksiri S, Mahasawat P (2005) In vitro platination of human breast cancer suppressor gene1 (BRCA1) by the anticancer drug carboplatin. Biochim Biophys Acta 1725:145–151. CrossRefGoogle Scholar
  37. 37.
    Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Method 25:402–408. CrossRefGoogle Scholar
  38. 38.
    Babak MV, Meier SM, Huber KVM et al (2015) Target profiling of an antimetastatic RAPTA agent by chemical proteomics: relevance to the mode of action. Chem Sci 6:2449–2456. CrossRefGoogle Scholar
  39. 39.
    Chakree K, Ovatlarnporn C, Dyson PJ, Ratanaphan A (2012) Altered DNA binding and amplification of human breast cancer suppressor gene BRCA1 induced by a novel antitumor compound, [Ru(η6-p-phenylethacrynate)Cl2(pta)]. Int J Mol Sci 13:13183–13202. CrossRefGoogle Scholar
  40. 40.
    Ratanaphan A, Temboot P, Dyson PJ (2010) In vitro ruthenation of human breast cancer suppress or gene 1 (BRCA1) by the antimetastasis compound RAPTA-C and its analogue CarboRAPTA-C. Chem Biodivers 7:1290–1302. CrossRefGoogle Scholar
  41. 41.
    Groessl M, Terenghi M, Casini A, Elviri L, Lobinski R, Dyson PJ (2010) Reactivity of anticancer metallodrugs with serum proteins: new insights from size exclusion chromatography-ICP-MS and ESI-MS. J Anal At Spectrom 25:305–313. CrossRefGoogle Scholar
  42. 42.
    Wolters DA, Stefanopoulou M, Dyson PJ, Groessl M (2012) Combination of metallomics and proteomics to study the effects of the metallodrug RAPTA-T on human cancer cells. Metallomics 4:1185–1196. CrossRefGoogle Scholar
  43. 43.
    Gavande NS, VanderVere-Carozza PS, Hinshaw HD et al (2016) DNA repair targeted therapy: the past or future of cancer treatment? Pharmacol Ther 160:65–83. CrossRefGoogle Scholar
  44. 44.
    Mendes F, Groessl M, Nazarov AA et al (2011) Metal-based inhibition of poly(ADP-ribose) polymerase-the guardian angel of DNA. J Med Chem 54:2196–2206. CrossRefGoogle Scholar
  45. 45.
    Temboot P, Lee RFS, Menin L, Patiny L, Dyson PJ, Ratanaphan A (2017) Biochemical and biophysical characterization of ruthenation of BRCA1 RING protein by RAPTA complexes and its E3 ubiquitin ligase activity. Biochem Biophys Res Commun 488:355–361. CrossRefGoogle Scholar
  46. 46.
    Adhireksan Z, Palermo G, Riedel T et al (2017) Allosteric cross-talk in chromatin can mediate drug-drug synergy. Nat Commun 8:14806–14817. CrossRefGoogle Scholar
  47. 47.
    Bergamo A, Gagliardi R, Scarcia V et al (1999) In vitro cell cycle arrest, in vivo action on solid metastasizing tumors, and host toxicity of the antimetastatic drug NAMI-A and cisplatin. J Pharmacol Exp Ther 289:559–564Google Scholar
  48. 48.
    Zajac M, Moneo MV, Carnero A, Benitez J, Martínez-Delgado B (2008) Mitotic catastrophe cell death induced by heat shock protein 90 inhibitor in BRCA1-deficient breast cancer cell lines. Mol Cancer Ther 7:2358–2366. CrossRefGoogle Scholar
  49. 49.
    Tassone P, Di Martino MT, Ventura M et al (2009) Loss of BRCA1 function increases the antitumor activity of cisplatin against human breast cancer xenografts in vivo. Cancer Biol Ther 8:648–653. CrossRefGoogle Scholar
  50. 50.
    Brabec V, Nováková O (2006) DNA binding mode of ruthenium complexes and relationship to tumor cell toxicity. Drug Resist Updat 9:111–122. CrossRefGoogle Scholar
  51. 51.
    James CR, Quinn JE, Mullan PB, Johnston PG, Harkin DP (2007) BRCA1, a potential predictive biomarker in the treatment of breast cancer. Oncologist 12:142–150. CrossRefGoogle Scholar
  52. 52.
    Lohse I, Borgida A, Cao P et al (2015) BRCA1 and BRCA2 mutations sensitize to chemotherapy in patient-derived pancreatic cancer xenografts. Br J Cancer 113:425–432. CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Laboratory of Pharmaceutical Biotechnology, Department of Pharmaceutical Chemistry, Faculty of Pharmaceutical SciencesPrince of Songkla UniversitySongkhlaThailand
  2. 2.Institute of Chemical Sciences and EngineeringSwiss Federal Institute of Technology Lausanne (EPFL)LausanneSwitzerland

Personalised recommendations