Cancer Chemotherapy and Pharmacology

, Volume 83, Issue 4, pp 681–692 | Cite as

In vitro and in vivo anticancer effects of two quinoline–platinum(II) complexes on human osteosarcoma models

  • Maria Carolina Ruiz
  • Agustina Resasco
  • Ana Laura Di Virgilio
  • Miguel Ayala
  • Isabel Cavaco
  • Silvia Cabrera
  • Jose Aleman
  • Ignacio Esteban LeónEmail author
Original Article


Platinum-based drugs, mainly cisplatin, are used for the treatment of several solid tumors such as OS. However, cisplatin treatment often results in the development of chemoresistance, leading therapeutic failure. We have previously reported that platinum complexes containing 8-hydroxyquinoline ligands have good antitumor activity against different cancer cell lines and with a different and better cytotoxic profile than cisplatin. Here, the anticancer properties of two different quinoline–platinum complexes [Pt(Cl)2(quinoline)(dmso)] (1) [PtCl(8-O-quinoline)(dmso)] (2) on in vitro (2D and 3D) and in vivo models (xenograft tumor of human osteosarcoma in mice) are presented. In this order, [PtCl(8-O-quinoline)(dmso)] (2) impaired cell viability to have a more pronounced antitumor effect than cisplatin on MG-63 osteosarcoma cells (IC50 4 µM vs. 39 µM). Besides, [PtCl(8-O-quinoline)(dmso)] (2) increased ROS production in a dose-manner response and this compound induced early and late apoptotic fractions of human osteosarcoma cells. Finally, [PtCl(8-O-quinoline)(dmso)] (2) decreased the cell viability of multicellular spheroids and reduced the tumor volume on athymic nude mice N:NIH(S) Fox1nu without inducing side effects. In this way, [PtCl(8-O-quinoline)(dmso)] (2) did not alter the normal cytoarchitecture of liver and kidney and the blood biomarkers (GPT, GOT, uremia, and creatinine) did not suffer modifications. Taken together, our data indicate that these compounds showed a better anticancer performance than cisplatin on in vitro and in vivo studies. These results showed the importance of chelation in the antitumor properties, suggesting that the [PtCl(8-O-quinoline)(dmso)] (2) might be a promising agent for the treatment of human osteosarcoma tumors resistant to cisplatin.


Platinum Osteosarcoma Spheroids Apoptosis 



SBE and IEL are members of the Research Carrer, CONICET, Argentina. JFCV and MCR have a fellowship from CONICET and ANPCyT, Argentina, respectively. We also gratefully acknowledge to Dra Ortiz Mayor from Hospital Padilla, Tucuman, Argentina, to help with the Histopathology studies. Moreover, the authors would like to thank MC. Bernal for her careful revision of the manuscript.


This work was partly supported by UNLP (11X/690), CONICET (PIP 00340), and ANPCyT (PICT 2014-2223) from Argentina and CTQ2015-64561-R from Spain.

Compliance with ethical standards

Conflict of interest

All the authors declare that they have no conflict of interest.

Ethical approval

The animal study was conducted in accordance with the Guide for the Care and Use of Laboratory Animals (National Research Council, USA 2002. Laboratory Animals. A.A Tuffery, 1995. London: John Wiley). All efforts were made to minimize animal suffering, to decrease the number of animals used, and to utilize possible alternatives to in vivo techniques.

Research involving human participants

This article does not contain any studies with human participants performed by any of the authors.

Supplementary material

280_2019_3773_MOESM1_ESM.tif (276 kb)
Figure SM1 Effect of cisplatin on the externalization of PS by flow cytometry in MG-63 cells. Cells were incubated with 25, 50 and 100 μM of cisplatin during 6 h (A). Graphical bars show the percentage of Annexin V (+) and Annexin V (+)/ PI (+) cells. Results are expressed as the mean ± SEM, n = 9, *significant differences vs. control (p < 0.01) (B). (C) Effect of complex 2 on the MMP by flow cytometry in MG-63 cells (TIF 276 KB)
280_2019_3773_MOESM2_ESM.tif (75 kb)
Figure SM2 Effects of cisplatin and compound 2 on survival rate. The Kaplan–Meier survival curve (A) Control vs cisplatin (B) Compound 2 vs cisplatin. The figure shows improvement of life span of xenograft-bearing mice treated compound 2 (6 mg/Kg) in comparison with cisplatin (6 mg/Kg) (n = 9 per group). Mice were treated as indicated in Figure 9 and were sacrificed throughout the study period upon reaching our study end point (TIF 74 KB)
280_2019_3773_MOESM3_ESM.tif (1.1 mb)
Figure SM3 Histopathology of tumor samples. Upper panel male tumor control samples, (A) Arrow, osteoid substance, Arrow-dash, atypical osteocytes cells, Circle, necrosis zone. (B) Tumor coagulative necrosis. Lower panel male tumor treatment samples. (C) Arrow, fibrotic tissue. (D) Circles, apoptotic focuses. Tumor samples were dissected by scalpel and stained with hematoxylin/eosin. Magnifications ×40, and ×100 are indicated (TIF 1161 KB)
280_2019_3773_MOESM4_ESM.tif (1.7 mb)
Figure SM4 Histopathology of the liver (A, B, D, E) and kidney samples (C, F). Upper panel male liver and kidney control samples, (A) Control liver architecture (B) Control liver Arrow: sinusoids, Dashed arrow: kupffer cells. (C) Control kidney architecture, Arrow: glomeruli, Dash arrow: tubules. Lower panel liver and kidney treatment samples (D) Treatment liver Arrow: microvacuolar (E) Treatment liver Arrow: Councilman hyaline bodies. (F) Treatment kidney, Arrow: glomeruli, Dash arrow: tubules. Samples were stained with hematoxylin/eosin. Magnifications ×40, and ×100 are indicated (TIF 1708 KB)


  1. 1.
    Kelland L (2007) The resurgence of platinum-based cancer chemotherapy. Nat Rev Cancer 7:573–584. CrossRefGoogle Scholar
  2. 2.
    Yamamoto N, Tsuchiya H (2013) Chemotherapy for osteosarcoma—where does it come from? What is it? Where is it going? Expert Opin Pharmacother 14:2183–2193. CrossRefPubMedGoogle Scholar
  3. 3.
    Gorlick R, Khanna C (2010) Osteosarcoma. J Bone Miner Res 25:683–691. CrossRefPubMedGoogle Scholar
  4. 4.
    Tsuchiya H, Tomita K, Mori Y et al (1999) Marginal excision for osteosarcoma with caffeine assisted chemotherapy. Clin Orthop Relat Res 358:27–35CrossRefGoogle Scholar
  5. 5.
    Bode AM, Dong Z (2007) The enigmatic effects of caffeine in cell cycle and cancer. Cancer Lett 247:26–39. CrossRefGoogle Scholar
  6. 6.
    Hattinger CM, Fanelli M, Tavanti E et al (2015) Advances in emerging drugs for osteosarcoma. Expert Opin Emerg Drugs. CrossRefPubMedGoogle Scholar
  7. 7.
    Durnali A, Alkis N, Cangur S et al (2013) Prognostic factors for teenage and adult patients with high-grade osteosarcoma: an analysis of 240 patients. Med Oncol 30:624. CrossRefPubMedGoogle Scholar
  8. 8.
    Holen I, Coleman RE (2010) Bisphosphonates as treatment of bone metastases. Curr Pharm Des 16:1262–1271CrossRefGoogle Scholar
  9. 9.
    McClung M, Harris ST, Miller PD et al (2013) Bisphosphonate therapy for osteoporosis: benefits, risks, and drug holiday. Am J Med 126:13–20. CrossRefPubMedGoogle Scholar
  10. 10.
    Xue Z, Lin M, Zhu J et al (2010) Platinum(II) compounds bearing bone-targeting group: synthesis, crystal structure and antitumor activity. Chem Commun 46:1212. CrossRefGoogle Scholar
  11. 11.
    Igarashi K, Yamamoto N, Hayashi K et al (2015) Effectiveness of two novel anionic and cationic platinum complexes in the treatment of osteosarcoma. Anticancer Agents Med Chem 15:390–399CrossRefPubMedGoogle Scholar
  12. 12.
    Johnstone TC, Suntharalingam K, Lippard SJ (2016) The next generation of platinum drugs: targeted Pt(II) agents, nanoparticle delivery, and Pt(IV) prodrugs. Chem Rev 116:3436–3486. CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Prachayasittikul V, Prachayasittikul S, Ruchirawat S, Prachayasittikul V (2013) 8-Hydroxyquinolines: a review of their metal chelating properties and medicinal applications. Drug Des Devel Ther 7:1157–1178. CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Afzal O, Kumar S, Haider MR et al (2015) A review on anticancer potential of bioactive heterocycle quinoline. Eur J Med Chem 97:871–910. CrossRefGoogle Scholar
  15. 15.
    Albrecht M, Fiege M, Osetska O (2008) 8-Hydroxyquinolines in metallosupramolecular chemistry. Coord Chem Rev 252:812–824. CrossRefGoogle Scholar
  16. 16.
    Martín Santos C, Cabrera S, Ríos-Luci C et al (2013) Novel clioquinol and its analogous platinum complexes: importance, role of the halogen substitution and the hydroxyl group of the ligand. Dalton Trans 42:13343–13348. CrossRefPubMedGoogle Scholar
  17. 17.
    Page H, Flood P, Reynaud EG (2013) Three-dimensional tissue cultures: current trends and beyond. Cell Tissue Res 352:123–131. CrossRefPubMedGoogle Scholar
  18. 18.
    Ho WY, Yeap SK, Ho CL et al (2012) Development of multicellular tumor spheroid (MCTS) culture from breast cancer cell and a high throughput screening method using the MTT assay. PLoS One 7:e44640. CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Friedrich J, Seidel C, Ebner R, Kunz-Schughart L (2009) Spheroid-based drug screen: considerations and practical approach. Nat Protoc 4:309–324. CrossRefPubMedGoogle Scholar
  20. 20.
    Alderden RA, Mellor HR, Modok S et al (2007) Elemental tomography of cancer-cell spheroids reveals incomplete uptake of both platinum(II) and platinum(IV) complexes. J Am Chem Soc 129:13400–13401. CrossRefPubMedGoogle Scholar
  21. 21.
    Modok S, Scott R, Alderden RA et al (2007) Transport kinetics of four- and six-coordinate platinum compounds in the multicell layer tumour model. Br J Cancer 97:194–200. CrossRefPubMedPubMedCentralGoogle Scholar
  22. 22.
    Price JH, Williamson AN, Schramm RF, Wayland BB (1972) Palladium(II) and platinum(II) alkyl sulfoxide complexes. Examples of sulfur-bonded, mixed sulfur- and oxygen-bonded, and totally oxygen-bonded complexes. Inorg Chem 11:1280–1284. CrossRefGoogle Scholar
  23. 23.
    Okajima T, Nakamura K, Zhang H et al (1992) Sensitive colorimetric bioassays for insulin-like growth factor (IGF) stimulation of cell proliferation and glucose consumption: use in studies of IGF analogs. Endocrinology 130:2201–2212. CrossRefPubMedGoogle Scholar
  24. 24.
    Mosmann T (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 65:55–63CrossRefPubMedPubMedCentralGoogle Scholar
  25. 25.
    Singh NP, McCoy MT, Tice RR, Schneider EL (1988) A simple technique for quantitation of low levels of DNA damage in individual cells. Exp Cell Res 175:184–191CrossRefGoogle Scholar
  26. 26.
    Butenko N, Isabel A, Nouri O et al (2009) DNA cleavage activity of V IV O (acac) 2 and derivatives. 103:622–632.
  27. 27.
    Bernadou J, Pratviel G, Bennis F et al (1989) Potassium monopersulfate and a water-soluble manganese porphyrin complex, [Mn(TMPyP)](OAc)5, as an efficient reagent for the oxidative cleavage of DNA. Biochemistry 28:7268–7275CrossRefPubMedGoogle Scholar
  28. 28.
    Leon IE, Di Virgilio a L, Porro V et al (2013) Antitumor properties of a vanadyl(IV) complex with the flavonoid chrysin [VO(chrysin)2EtOH]2 in a human osteosarcoma model: the role of oxidative stress and apoptosis. Dalton Trans 42:11868–11880. CrossRefPubMedGoogle Scholar
  29. 29.
    León IE, Cadavid-Vargas JF, Resasco A et al (2016) In vitro and in vivo antitumor effects of the VO-chrysin complex on a new three-dimensional osteosarcoma spheroids model and a xenograft tumor in mice. JBIC J Biol Inorg Chem 21:1009–1020. CrossRefPubMedGoogle Scholar
  30. 30.
    ElBayoumi TA, Torchilin VP (2009) Tumor-targeted nanomedicines: enhanced antitumor efficacy in vivo of doxorubicin-loaded, long-circulating liposomes modified with cancer-specific monoclonal antibody. Clin Cancer Res 15:1973–1980. CrossRefPubMedPubMedCentralGoogle Scholar
  31. 31.
    Pregosin PS (1982) Platinum-195 nuclear magnetic resonance. Coord Chem Rev 44:247–291. CrossRefGoogle Scholar
  32. 32.
    León IE, Di Virgilio AL, Barrio DA et al (2012) Hydroxylamido-amino acid complexes of oxovanadium(V). Toxicological study in cell culture and in a zebrafish model. Metallomics 4:1287–1296. CrossRefPubMedGoogle Scholar
  33. 33.
    Flocke LS, Trondl R, Jakupec MA, Keppler BK (2016) Molecular mode of action of NKP-1339—a clinically investigated ruthenium-based drug—involves ER- and ROS-related effects in colon carcinoma cell lines. Invest New Drugs 34:261–268. CrossRefPubMedPubMedCentralGoogle Scholar
  34. 34.
    Terenzi A, Pirker C, Keppler BK, Berger W (2016) Anticancer metal drugs and immunogenic cell death. J Inorg Biochem 165:71–79. CrossRefPubMedGoogle Scholar
  35. 35.
    Kamogashira T, Fujimoto C, Yamasoba T (2015) Reactive oxygen species, apoptosis, and mitochondrial dysfunction in hearing loss. Biomed Res Int 2015:617207. CrossRefPubMedPubMedCentralGoogle Scholar
  36. 36.
    Slimen IB, Najar T, Ghram A et al (2014) Reactive oxygen species, heat stress and oxidative-induced mitochondrial damage. A review. Int J Hyperthermia 30:513–523. CrossRefPubMedGoogle Scholar
  37. 37.
    Herr I, Debatin KM (2001) Cellular stress response and apoptosis in cancer therapy. Blood 98:2603–2614CrossRefPubMedGoogle Scholar
  38. 38.
    Mayer B, Oberbauer R (2003) Mitochondrial regulation of apoptosis. News Physiol Sci 18:89–94PubMedGoogle Scholar
  39. 39.
    Walzl A, Unger C, Kramer N et al (2014) The resazurin reduction assay can distinguish cytotoxic from cytostatic compounds in spheroid screening assays. J Biomol Screen 19:1047–1059. CrossRefPubMedGoogle Scholar
  40. 40.
    Chen JL, Steele TWJ, Stuckey DC (2015) Modeling and application of a rapid fluorescence-based assay for biotoxicity in anaerobic digestion. Environ Sci Technol 49:13463–13471. CrossRefPubMedGoogle Scholar
  41. 41.
    Schreiber-Brynzak E, Klapproth E, Unger C et al (2015) Three-dimensional and co-culture models for preclinical evaluation of metal-based anticancer drugs. Invest New Drugs 33:835–847. CrossRefPubMedGoogle Scholar
  42. 42.
    Shahid F, Farooqui Z, Khan F (2018) Cisplatin-induced gastrointestinal toxicity: an update on possible mechanisms and on available gastroprotective strategies. Eur J Pharmacol 827:49–57. CrossRefPubMedGoogle Scholar
  43. 43.
    Perše M, Večerić-Haler Ž (2018) Cisplatin-induced rodent model of kidney injury: characteristics and challenges. Biomed Res Int 2018:1–29. CrossRefGoogle Scholar
  44. 44.
    Qin Q-P, Chen Z-F, Qin J-L et al (2015) Studies on antitumor mechanism of two planar platinum(II) complexes with 8-hydroxyquinoline: synthesis, characterization, cytotoxicity, cell cycle and apoptosis. Eur J Med Chem 92:302–313. CrossRefPubMedGoogle Scholar
  45. 45.
    Dilruba S, Kalayda GV (2016) Platinum-based drugs: past, present and future. Cancer Chemother Pharmacol 77:1103–1124. CrossRefPubMedGoogle Scholar
  46. 46.
    Roberts NB, Wadajkar AS, Winkles JA et al (2016) Repurposing platinum-based chemotherapies for multi-modal treatment of glioblastoma. Oncoimmunology 5:e1208876. CrossRefPubMedPubMedCentralGoogle Scholar
  47. 47.
    Ruggiero A, Trombatore G, Triarico S et al (2013) Platinum compounds in children with cancer. Anticancer Drugs 24:1007–1019. CrossRefPubMedGoogle Scholar
  48. 48.
    Brozovic A, Ambriović-Ristov A, Osmak M (2010) The relationship between cisplatin-induced reactive oxygen species, glutathione, and BCL-2 and resistance to cisplatin. Crit Rev Toxicol 40:347–359. CrossRefGoogle Scholar
  49. 49.
    Fujii H, Honoki K, Tsujiuchi T et al (2009) Sphere-forming stem-like cell populations with drug resistance in human sarcoma cell lines. Int J Oncol 34:1381–1386PubMedGoogle Scholar
  50. 50.
    Langdon SP (2012) Animal modeling of cancer pathology and studying tumor response to therapy. Curr Drug Targets 13:1535–1547CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Chair of Pathologic Biochemistry, Exact School SciencesNational University of La PlataLa PlataArgentina
  2. 2.Inorganic Chemistry Center (CONICET-UNLP) Exact School SciencesNational University of La PlataLa PlataArgentina
  3. 3.Lab Experimental Animals, Veterinary School SciencesNational University of La PlataLa PlataArgentina
  4. 4.Chemistry, Biochemistry and Pharmacy DepartmentAlgarve UniversityFaroPortugal
  5. 5.Inorganic Chemistry DepartmentAutonomous University of MadridMadridSpain
  6. 6.Organic Chemistry DepartmentUniversidad Autónoma de MadridMadridSpain

Personalised recommendations