The Journal of Membrane Biology

, Volume 247, Issue 12, pp 1239–1251 | Cite as

Modulation of Activity of Known Cytotoxic Ruthenium(III) Compound (KP418) with Hampered Transmembrane Transport in Electrochemotherapy In Vitro and In Vivo

  • Rosana Hudej
  • Damijan Miklavcic
  • Maja Cemazar
  • Vesna Todorovic
  • Gregor Sersa
  • Alberta Bergamo
  • Gianni Sava
  • Anze Martincic
  • Janez Scancar
  • Bernhard K. Keppler
  • Iztok Turel
Article

Abstract

To increase electrochemotherapy (ECT) applicability, the effectiveness of new drugs is being tested in combination with electroporation. Among them two ruthenium(III) compounds, (imH)[trans-RuCl4(im)(DMSO-S)] (NAMI-A) and Na[trans-RuCl4(ind)2] (KP1339), proved to possess increased antitumor effectiveness when combined with electroporation. The objective of our experimental work was to determine influence of electroporation on the cytotoxic and antitumor effect of a ruthenium(III) compound with hampered transmembrane transport, (imH)[trans-RuCl4(im)2] (KP418) in vitro and in vivo and to determine changes in metastatic potential of cells after ECT with KP418 in vitro. In addition, platinum compound cisplatin (CDDP) and ruthenium(III) compound NAMI-A were included in the experiments as reference compounds. Our results show that electroporation leads to increased cellular accumulation and cytotoxicity of KP418 in murine melanoma cell lines with low and high metastatic potential, B16-F1 and B16-F10, but not in murine fibrosarcoma cell line SA-1 in vitro which is probably due to variable effectiveness of ECT in different cell lines and tumors. Electroporation does not potentiate the cytotoxicity of KP418 as prominently as the cytotoxicity of CDDP. We also showed that the metastatic potential of cells which survived ECT with KP418 or NAMI-A does not change in vitro: resistance to detachment, invasiveness, and re-adhesion of cells after ECT is not affected. Experiments in murine tumor models B16-F1 and SA-1 showed that ECT with KP418 does not have any antitumor effect while ECT with CDDP induces significant dose-dependent tumor growth delay in the two tumor models used in vivo.

Keywords

KP418 Electrochemotherapy Ruthenium Metastatic potential In vitro In vivo 

References

  1. Antonarakis ES, Emadi A (2010) Ruthenium-based chemotherapeutics: are they ready for prime time? Cancer Chemoth Pharm 66:1–9CrossRefGoogle Scholar
  2. Bergamo A, Masi A, Jakupec MA, Keppler BK, Sava G (2009) Inhibitory effect of the ruthenium complex KP1019 in models of mammary cancer cell migration and invasion. Met-Based Drugs. doi:10.1155/2009/681270 PubMedCentralPubMedGoogle Scholar
  3. Berger MR, Garzon FT, Keppler BK, Schmähl D (1989) Efficacy of new ruthenium complexes against chemically induced autochthonous colorectal carcinoma in rats. Anticancer Res 9:761–766PubMedGoogle Scholar
  4. Bicek A, Turel I, Kanduser M, Miklavcic D (2007) Combined therapy of the antimetastatic compound NAMI-A and electroporation on B16F1 tumour cells in vitro. Bioelectrochemistry 71:113–117CrossRefPubMedGoogle Scholar
  5. Brincker H (1993) Direct intratumoral chemotherapy. Crit Rev Oncol Hemat 15:91–98CrossRefGoogle Scholar
  6. Cemazar M, Miklavcic D, Vodovnik L, Jarm T, Rudolf Z, Stabuc B, Cufer T, Sersa G (1995) Improved therapeutic effect of electrochemotherapy with cisplatin by intratumoral drug administration and changing of electrode orientation for electropermeabilization on EAT tumor model in mice. Radiol Oncol 29:121–127Google Scholar
  7. Cemazar M, Jarm T, Miklavcic D, Macek Lebar A, Ihan A, Kopitar NA, Sersa G (1998) Effect of electric field intensity on electropermeabilization and electrosensitivity of various tumor-cell lines in vitro. Electro Magnetobiol 17:263–272CrossRefGoogle Scholar
  8. Cemazar M, Miklavcic D, Scancar J, Dolzan V, Golouh R, Sersa G (1999) Increased platinum accumulation in SA-1 tumor cells after in vivo electrochemotherapy with cisplatin. Br J Cancer 79:1386–1391PubMedCentralCrossRefPubMedGoogle Scholar
  9. Cemazar M, Parkins CS, Holder AL, Chaplin DJ, Tozer GM, Sersa G (2001) Electroporation of human microvascular endothelial cells: evidence for an anti-vascular mechanism of electrochemotherapy. Brit J Cancer 84:565–570PubMedCentralCrossRefPubMedGoogle Scholar
  10. Cepeda V, Fuertes MA, Castilla J, Alonso C, Quevedo C, Pérez JM (2007) Biochemical machanisms of cisplatin cytotoxicity. Anticancer Agents Med Chem 7:3–18CrossRefPubMedGoogle Scholar
  11. Corovic S, Lackovic I, Sustaric P, Sustar T, Rodic T, Miklavcic D (2013) Modeling of electric field distribution in tissues during electroporation. Biomed Eng Online 12:16PubMedCentralCrossRefPubMedGoogle Scholar
  12. Edhemovic I, Gadzijev EM, Brecelj E, Miklavcic D, Kos B, Zupanic A, Mali B, Jarm T, Pavliha D, Marcan M, Gasljevic G, Gorjup V, Music M, Pecnik Vavpotic T, Cemazar M, Snoj M, Sersa G (2011) Electrochemotherapy: a new technological approach in treatment of metastases in the liver. Technol Cancer Res Treat 10:475–485PubMedCentralPubMedGoogle Scholar
  13. Gava B, Zorzet P, Spessotto P, Cocchietto M, Sava G (2006) Inhibition of B16 melanoma metastases with the ruthenium complex imidazolium trans-imidazoledimethylsulfoxide-tetrachlororuthenate and down-regulation of tumor cell invasion. J Pharmacol Exp Ther 317(1):284–291CrossRefPubMedGoogle Scholar
  14. Haberl S, Miklavcic D, Sersa G, Frey W, Rubinsky B (2013) Cell membrane electroporation—Part 2: The applications. IEEE Electr Insul M 29(1):29–37CrossRefGoogle Scholar
  15. Hartinger CG, Jakupec MA, Zorbas-Seifried S, Groessl M, Egger A, Berger W, Zorbas H, Dyson PJ, Keppler BK (2008) KP1019, a new redox-active anticancer agent-preclinical development and results of a clinical phase I study in tumor patients. Chem Biodivers 5:2140–2155CrossRefPubMedGoogle Scholar
  16. Heffeter P, Böck K, Atil B, Hoda MAR, Körner W, Bartel C, Jungwirth U, Keppler BK, Micksche M, Berger W, Koellensperger G (2010) Intracellular protein binding patterns of the anticancer ruthenium drugs KP1019 and KP1339. J Biol Inorg Chem 15:737–748PubMedCentralCrossRefPubMedGoogle Scholar
  17. Heller R, Gilbert R, Jaroszeski MJ (2000) Clinical trials for solid tumors using electrochemotherapy. In: Electrochemotherapy, Electrogenetherapy, and Transdermal Drug Delivery. Electrically Mediated Delivery of Molecules to Cells. Jaroszeski MJ, Heller R and Gilbert R (eds.). New Jersey: Humana Press, pp. 137-156Google Scholar
  18. Hudej R, Turel I, Kanduser M, Scancar J, Kranjc S, Sersa G, Miklavcic D, Jakupec MA, Keppler BK, Cemazar M (2010) The influence of electroporation on cytotoxicity of anticancer ruthenium(III) complex KP1339 in vitro and in vivo. Anticancer Res 30:2055–2064PubMedGoogle Scholar
  19. Hudej R, Kljun J, Kandioller W, Repnik U, Turk B, Hartinger CG, Keppler BK, Miklavcic D, Turel I (2012) Synthesis and Biological Evaluation of the Thionated Antibacterial Agent Nalidixic Acid and its Organoruthenium(II) Complex. Organometallics 31:5867–5874CrossRefGoogle Scholar
  20. Jaroszeski MJ, Dang V, Pottinger C, Hickey J, Gilbert R, Heller R (2000) Toxicity of anticancer agents mediated by electroporation in vitro. Anti-Cancer Drug 11:201–208CrossRefGoogle Scholar
  21. Kapitza S, Pongratz M, Jakupec MA, Heffeter P, Berger W, Lackinger L, Keppler BK, Marian B (2005) Heterocyclic complexes of ruthenium(III) induce apoptosis in colorectal carcinoma cells. J Cancer Res Clin Oncol 131:101–110CrossRefPubMedGoogle Scholar
  22. Kersten L, Bräunlich H, Keppler BK, Gliesing C, Wendelin M, Westphal J (1998) Comparative nephrotoxicity of some antitumour-active platinum and ruthenium complexes in rats. J Appl Toxicol 18:93–101CrossRefPubMedGoogle Scholar
  23. Kljun J, Petricek S, Zigon D, Hudej R, Miklavcic D, Turel I (2010) Synthesis and Characterization of Novel Ruthenium(III) Complexes with Histamine. Bioinorg Chem Appl 183097:1–6CrossRefGoogle Scholar
  24. Mali B, Jarm T, Snoj M, Sersa G, Miklavcic D (2013a) Antitumor effectiveness of electrochemotherapy: a systematic review and meta-analysis. Eur J SurgOncol 39:4–16CrossRefGoogle Scholar
  25. Mali B, Miklavcic D, Campana LG, Cemazar M, Sersa G, Snoj M, Jarm T (2013b) Tumor size and effectiveness of electrochemotherapy. Radiol Oncol 47:32–41PubMedCentralCrossRefPubMedGoogle Scholar
  26. Marty M, Sersa G, Garbay JR, Gehl J, Collins CG, Snoj M, Billard V, Geertsen PF, Larkin JO, Miklavcic D (2006) Electrochemotherapy—an easy, highly effective and safe treatment of cutaneous and subcutaneous metastases: results of ESOPE (European Standard Operating Procedures of Electrochemotherapy) study. Eur J Cancer (Suppl 4): 3-13Google Scholar
  27. Miklavcic D, Snoj M, Zupanic A, Kos B, Cemazar M, Kropivnik M, Bracko M, Pecnik T, Gadzijev E, Sersa G (2010) Towards treatment planning and treatment of deep-seated solid tumors by electrochemotherapy. Biomed Eng Online 9:10PubMedCentralCrossRefPubMedGoogle Scholar
  28. Miklavcic D, Sersa G, Brecelj E, Gehl J, Soden D, Bianchi G, Ruggieri P, Rossi CR, Campana LG, Jarm T (2012) Electrochemotherapy: technological advancements for efficient electroporation-based treatment of internal tumors. Med Biol Eng Comput 50:1213–1225PubMedCentralCrossRefPubMedGoogle Scholar
  29. Miklavcic D, Mali B, Kos B, Heller R, Sersa G (2014) Electrochemotherapy: from the drawing board into medical practice. Biomed Eng Online 13:29PubMedCentralCrossRefPubMedGoogle Scholar
  30. Mir LM, Gehl J, Sersa G, Collins CG, Garbay JR, Billard V, Geertsen PF, Rudolf Z, O’Sullivan GC, Marty M (2006) Standard operating procedures of the electrochemotherapy: instructions for the use of bleomycin or CDDP administered either systemically or locally and electric pulses delivered by the CliniporatorTM by means of invasive or non-invasive electrodes. Eur J Cancer 4:14–25CrossRefGoogle Scholar
  31. Orlowski S, Belehradek J Jr, Paoletti C, Mir LM (1988) Transient electropermeabilization of cells in culture: increase of the cytotoxicity of anticancer drugs. Biochem Pharmacol 37(24):4727–4733CrossRefPubMedGoogle Scholar
  32. Pavliha D, Kos B, Marcan M, Zupanic A, Sersa G, Miklavcic D (2013a) Planning of electroporation-based treatments using web-based treatment-planning software. J Membrane Biol 246:833–842CrossRefGoogle Scholar
  33. Pavliha D, Music MM, Sersa G, Miklavcic D (2013b) Electroporation-based treatment planning for deep-seated tumors based on automatic liver segmentation of MRI images. PLoS ONE 8(8):e69068PubMedCentralCrossRefPubMedGoogle Scholar
  34. Pucihar G, Kotnik T, Valic B, Miklavcic D (2006) Numerical determination of transmembrane voltage induced on irregularly shaped cells. Annals Biomed Eng 34:642–652CrossRefGoogle Scholar
  35. Sava G, Zorzet S, Turrin C, Vita F, Soranzo MR, Zabucchi G, Cocchietto M, Bergamo A, DiGiovine S, Pezzoni G, Sartor L, Garbisa S (2003) Dual action of NAMI-A in inhibition of solid tumor metastasis: selective targeting of metastatic cells and binding to collagen. Clin Cancer Res 9:1898–1905PubMedGoogle Scholar
  36. Seelig MH, Berger MR, Keppler BK (1992) Antineoplastic activity of three ruthenium derivatives against chemically induced colorectal carcinoma in rats. J Cancer Res Clin Oncol 118:195–200CrossRefPubMedGoogle Scholar
  37. Sersa G, Cemazar M, Miklavcic D, Mir LM (1994) Electrochemotherapy: variable anti-tumor effect on different tumor models. Bioelectrochem Bioenerg 35:23–27CrossRefGoogle Scholar
  38. Sersa G, Cemazar M, Miklavcic D (1995) Antitumor effectiveness of electrochemotherapy with cis-diamminedichloroplatinum(II) in mice. Cancer Res 55:3450–3455PubMedGoogle Scholar
  39. Sersa G, Krzic M, Sentjurc M, Ivanusa T, Beravs K, Kotnik V, Coer A, Swartz HM, Cemazar M (2002) Reduced blood flow and oxygenation in SA-1 tumours after electrochemotherapy with cisplatin. Brit J Cancer. 87:1047–1054PubMedCentralCrossRefPubMedGoogle Scholar
  40. Sersa G, Miklavcic D, Cemazar M, Rudolf Z, Pucihar G, Snoj M (2008) Electrochemotherapy in treatment of tumours. Eur J Surg Oncol 34:232–240CrossRefPubMedGoogle Scholar
  41. Speelmans G, Sips WHHM, Grisel RJH, Staffhorst RWHM, Fichtinger-Schepman AMJ, Reedijk J, de Kruijff B (1996) The interaction of the anti-cancer drug cisplatin with phospholipids is specific for negatively charged phospholipids and takes place at low chloride ion concentration. Biochim Biophys Acta 1283:60–66CrossRefPubMedGoogle Scholar
  42. Spugnini EP, Citro G, Porrello A (2005) Rational design of new electrodes for electrochemotherapy. J ExpClin Cancer Res 24(2):245–254Google Scholar
  43. Todorovic V, Sersa G, Mlakar V, Glavac D, Flisar K, Cemazar M (2011) Metastatic potential of melanoma cells is not affected by electrochemotherapy. Melanoma Res 21:196–205CrossRefPubMedGoogle Scholar
  44. Todorovic V, Sersa G, Mlakar V, Glavac D, Cemazar M (2012) Assessment of the tumourigenic and metastatic properties of SK-MEL28 melanoma cells surviving electrochemotherapy with bleomycin. Radiol Oncol 46(1):32–45PubMedCentralCrossRefPubMedGoogle Scholar
  45. Zorzet S, Bergamo A, Cocchietto M, Sorc A, Gava B, Alessio E, Iengo E, Sava G (2000) Lack of in vitro cytotoxicity, associated to increased G2-M cell fraction and inhibition of matrigel invasion, may predict in vivo-selective antimetastasis activity of ruthenium complexes. JPET 295(3):927–933Google Scholar

Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Rosana Hudej
    • 1
    • 2
  • Damijan Miklavcic
    • 1
  • Maja Cemazar
    • 3
    • 4
  • Vesna Todorovic
    • 3
  • Gregor Sersa
    • 3
  • Alberta Bergamo
    • 5
  • Gianni Sava
    • 5
    • 6
  • Anze Martincic
    • 7
  • Janez Scancar
    • 7
  • Bernhard K. Keppler
    • 8
  • Iztok Turel
    • 9
  1. 1.Faculty of Electrical EngineeringUniversity of LjubljanaLjubljanaSlovenia
  2. 2.BIA Separations d.o.o.AjdovscinaSlovenia
  3. 3.Institute of Oncology LjubljanaLjubljanaSlovenia
  4. 4.Faculty of Health SciencesUniversity of PrimorskaIzolaSlovenia
  5. 5.Callerio FoundationTriesteItaly
  6. 6.Department of biomedical ScienceUniversity of TriesteTriesteItaly
  7. 7.Jozef Stefan InstituteLjubljanaSlovenia
  8. 8.Institute of Inorganic ChemistryUniversity of ViennaViennaAustria
  9. 9.Faculty of Chemistry and Chemical TechnologyUniversity of LjubljanaLjubljanaSlovenia

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