Skip to main content

Part of the book series: Methods in Molecular Medicine ((MIMM,volume 37))

Abstract

Antitumor electrochemotherapy is a treatment of solid tumors which combines a cytotoxic nonpermeant drug, like bleomycin, with locally delivered permeabilizing electric pulses (13). More generally, a new form of vectorization is achieved by the combination of nonpermeant molecules with intracellular targets and of a physical perturbation that locally permeabilizes the cells. This vectorization does not require chemical, biochemical or biological modifications of the targeted compound, since the modification is performed on the target cells. A very convenient way to transiently permeabilize the cells is the use of appropriate electric pulses (short and intense squarewave electric pulses) that are not cytotoxic by themselves (1). These electric pulses reversibly permeabilize the electropulsed cells. Consequently, they allow increased drug delivery inside cells, particularly in the case of drugs for which the plasma membrane is a barrier that limits their access inside the cell [termed nonpermeant drugs] (4). As illustrated in the various protocols reported in this volume, electrochemotherapy using bleomycin is efficient to eradicate subcutaneously transplanted and spontaneous small tumors in mice and rats as well as experimental internal tumors transplanted in rat brain or in rabbit or rat liver. All the clinical trials (3,510) confirm the efficacy of this new therapeutical approach based on an original way to deliver nonpermeant cytotoxic drugs

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

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 129.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 169.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Mir, L. M., Orlowski, S., Belehradek, J. Jr., and Paoletti, C. (1991) Electrochemotherapy: potentiation of antitumour effect of bleomycin by local electric pulses. Eur. J. Cancer 27, 68–72.

    Article  PubMed  CAS  Google Scholar 

  2. Belehradek, J. Jr., Orlowski, S., Poddevin, B., Paoletti, C., and Mir, L. M. (1991) Electrochemotherapy of spontaneous mammary tumours in mice. Eur. J. Cancer 27, 73–76.

    Article  PubMed  Google Scholar 

  3. Belehradek, M., Domenge, C., Luboinski, B., Orlowski, S., Belehradek, J. Jr., and Mir L. M. (1993) Electrochemotherapy, a new antitumour treatment: First clinical phase I-II trial. Cancer 72, 3694–3700.

    Article  PubMed  CAS  Google Scholar 

  4. Mir, L. M., Banoun, H., and Paoletti, C. (1988) Introduction of definite amounts of nonpermeant molecules into living cells after electropermeabilization: Direct access to the cytosol. Exp. Cell Res. 175, 15–25.

    Article  PubMed  CAS  Google Scholar 

  5. Domenge, C., Orlowski, S., Luboinski, B., De Baere, T., Schwaab, G., Belehradek, J. Jr., and Mir, L. M. (1996) Antitumor electrochemotherapy: New advances in the clinical protocols. Cancer 77, 956–963.

    Article  PubMed  CAS  Google Scholar 

  6. Heller, R., Jaroszeski, M., Perrott, R., Glass, L. F., Puleo, C., Messina, J. L., Rapaport, D. P., DeConti, R. C., Fenske, N. A., Gilbert, R., Mir, L. M., and Reintgen, D. S. (1996) Phase I/II for the treatment of cutaneous and subcutaneous tumors using electrochemotherapy. Cancer 77, 964–972.

    Article  PubMed  CAS  Google Scholar 

  7. Rudolf, Z., Štabuc, B., Čemažar, M., Miklavčič, D., Vodovnik, L., and Serša, G. (1996) Electrochemotherapy with bleomycin: The first clinical experience in malignant melanoma patients. Radiol. Oncol. 27, 229–235.

    Google Scholar 

  8. Glass, L. F., Fenske, N. A., Jaroszeski, M. J., Perrott, R., Harvey, D. T., Reintgen, D. S., and Heller, R. (1996) Bleomycin-mediated electrochemotherapy of basal cell carcinoma. J. Am. Acad. Dermatol. 34, 82–86.

    Article  PubMed  CAS  Google Scholar 

  9. Glass, L. F., Jaroszeski, M., Gilbert, R., Reintgen, D. S., and Heller, R. (1997) Intralesional bleomycin-mediated electrochemotherapy in 20 patients with basal cell carcinoma. J. Am. Acad. Dermatol. 37, 596–599.

    Article  PubMed  CAS  Google Scholar 

  10. Mir, L. M., Glass, F. L., Serša, G., Teissié, J., Domenge, C., Miklavčič, D., Jaroszeski, M. J., Orlowski, S., Reintgen, D. S., Rudolf, Z., Belehradek, M., Gilbert, R., Rols, M. P., Belehradek, J. Jr, Bachaud, J. M., DeConti, R., Štabuc, B., Čemažar, M., Coninx, P., and Heller, R. (1998) Effective treatment of cutaneous and subcutaneous malignant tumours by electrochemotherapy. Br. J. Cancer 77, 2336–2342.

    Article  PubMed  CAS  Google Scholar 

  11. Zimmermann, U., Pilwat, G., and Riemann, F. (1974) Dielectric breakdown of cell membranes. Biophys. J. 14, 881–899.

    Article  PubMed  CAS  Google Scholar 

  12. Zimmermann, U. (1982) Electric field-mediated fusion and related electrical phenomena. Biochim. Biophys. Acta 694, 227–277.

    PubMed  CAS  Google Scholar 

  13. Neumann, E. (1992) Membrane electroporation and direct gene transfer. Bioelectrochem. Bioenerg. 28, 247–267.

    Article  CAS  Google Scholar 

  14. Neumann, E. and Rosenheck, K. (1972) Permeability changes induced by electric impulses in vesicular membranes. J. Membr. Biol. 10, 279–290.

    Article  PubMed  CAS  Google Scholar 

  15. Kinosita, K. Jr. and Tsong, T. Y. (1977) Formation and resealing of pores of controlled sizes in human erythrocyte membrane. Nature 268, 438–441.

    Article  PubMed  Google Scholar 

  16. Gross, D., Loew, L. M., and Webb, W. W. (1986) Optical imaging of cell membrane potential changes induced by applied electric fields. Biophys. J. 50, 339–348.

    Article  PubMed  CAS  Google Scholar 

  17. Belehradek, J. Jr., Orlowski, S., Ramirez, L. H., Pron, G., Poddevin, B., and Mir, L. M. (1994) Electropermeabilization of cells in tissues assessed by the qualitative and quantitative electroloading of bleomycin. Biochim. Biophys. Acta 1190, 155–163.

    Article  PubMed  CAS  Google Scholar 

  18. Okino, M. and Mohri, H. (1987) Effects of a high voltage electrical impulse and an anticancer drug on in vivo growing tumors. Jpn. J. Cancer Res. (Gann) 78, 1319–1321.

    PubMed  CAS  Google Scholar 

  19. Djuzenova, C. S., Zimmermann, U., Frank, H., Sukhorukov, V. L., Richter, E., and Fuhr, G. (1996) Effect of medium conductivity and composition on the uptake of propidium iodide into electropermeabilized myeloma cells. Biochim. Biophys. Acta 1284, 143–152.

    Article  PubMed  Google Scholar 

  20. Rols, M-P. and Teissié, J. (1990) Electropermeabilization of mammalian cells. Quantitative analysis of the phenomenon. Biophys. J. 58, 1089–1098.

    Article  PubMed  CAS  Google Scholar 

  21. Poddevin, B., Orlowski, S., Belehradek, J. Jr., and Mior, L. M. (1991) Very high cytotoxicity of bleomycin introduced into the cytosol of cells in culture. Biochem. Pharmacol. 42 (Suppl.), 67-75.

    Google Scholar 

  22. Gehl, J., Sørensen, T. H., Nielsen, K., Raskmark, P., Nielsen, S. L., Skovsgaard, T. and Mir, L. M. (1999) In vivo electroporation of skeletal muscle: threshold, efficacy and relation to electric field distribution. Biochim. Biophys. Acta 1428, 233–240.

    CAS  Google Scholar 

  23. Gehl, J. and Mir, L. M. (1999) Determination of optimal parameters for in vivo gene transfer by electroporation, using a rapid in vivo test for cell permeabilization. Biochem. Biophys. Res. Comm. 261, 377–380.

    Article  PubMed  CAS  Google Scholar 

  24. Neumann, E., Schaefer-Ridder, M., Wang, Y., and Hofschneider, P. H. (1982) Gene transfer into mouse lyoma cells by electroporation in high electric fields. EMBO J. 1, 841–845.

    PubMed  CAS  Google Scholar 

  25. Chang, D. C., Gao, P.-Q., and Maxwell, B. L. (1991) High efficiency gene transfection by electroporation using a radio-frequency electric field. Biochim. Biophys. Acta 1092, 153–160.

    Article  PubMed  CAS  Google Scholar 

  26. Gehl, J., Skovsgaard, T., and Mir, L. M. (1998) Enhancement of cytotoxicity by electropermeabilization: An improved method for screening drugs. Anti-Cancer Drugs 9, 319–325.

    Article  PubMed  CAS  Google Scholar 

  27. Takahashi, M., Furukawa, T., Saitoh, H., Aoki, A., Koike, T., Moriyama, Y., and Shibata, A. (1991) Gene transfer into human leukemia cell lines by electroThe Basis of Electrochemotherapy 115 poration: Experience with exponentially decaying and square-wave pulses. Leuk. Res. 15, 507–513.

    Article  PubMed  CAS  Google Scholar 

  28. Okino, M., Tomie, H., Kanesada, H., Marumoto, M., Esato, K., and Suzuki, H. (1992) Optimal electric conditions in electrical impulse chemotherapy. Jpn. J. Cancer Res. (Gann) 83, 1095–1101.

    PubMed  CAS  Google Scholar 

  29. Orlowski, S., Belehradek, J. Jr., Paoletti, C., and Mir, L. M. (1988) Transient electropermeabilization of cells in culture: Increase of the cytotoxicity of anticancer drugs. Biochem. Pharmacol. 37, 4727–4733.

    Article  PubMed  CAS  Google Scholar 

  30. Rols, M-P. and Teissié, J. (1989) Ionic-strength modulation of electrically-induced permeabilization and associated fusion of mammalian cells. Eur. J. Biochem. 179, 109–115.

    Article  PubMed  CAS  Google Scholar 

  31. Serša, G., Čemažar, M., Šemrov, D., and Miklavčič, D. (1996) Changing the electrode orientation improves the efficacy of electrochemotherapy of solid tumors in mice. Bioelectrochem. Bioenerg. 39, 61–66.

    Article  Google Scholar 

  32. Mir, L. M., Tounekti, O., and Orlowski, S. (1996) Bleomycin: Revival of an old drug. Gen. Pharmacol. 27, 745–748.

    PubMed  CAS  Google Scholar 

  33. Pron, G., Belehradek, J. Jr., Orlowski, S., and Mir, L. M. (1994) Involvement of the membrane bleomycin-binding sites in the bleomycin cytotoxicity. Biochem. Pharmacol. 48, 301–310.

    Article  PubMed  CAS  Google Scholar 

  34. Pron, G., Belehradek, J. Jr., and Mir, L. M. (1993) Identification of a plasma membrane protein that specifically binds bleomycin. Biochem. Biophys. Res. Commun. 194, 333–337.

    Article  CAS  Google Scholar 

  35. Lazo, J. S., Schisselbauer, J. C., Herring, G. M., and Kennedy, K. A. (1990) Involvement of the cellular vacuolar system with the cytotoxicity of bleomycinlike agents. Cancer Commun. 2, 81–86.

    PubMed  CAS  Google Scholar 

  36. Pron, G., Mahrour, N., Orlowski, S., Tounekti, O., Poddevin, B., Belehradek, J. Jr., and Mir, L. M. (1999) Internalisation of the bleomycin molecules responsible for bleomycin toxicity: A receptor-mediated endocytosis mechanism. Biochem. Pharmacol. 57, 45–56.

    Article  PubMed  CAS  Google Scholar 

  37. Orlowski, S. and Mir, L. M. (1993) Cell electropermeabilization: A new tool for biochemical and pharmacological studies. Biochim. Biophys. Acta 1154, 51–63.

    PubMed  CAS  Google Scholar 

  38. Tounekti, O., Pron, G., Belehradek, J. Jr., and Mir, L. M. (1993) Bleomycin, an apoptosis-mimetic drug that induces two types of cell death depending on the number of molecules internalized. Cancer Res. 53, 5462–5469.

    PubMed  CAS  Google Scholar 

  39. Tounekti, O., Belehradek, J. Jr., and Mir, L. M. (1995) Relationships between DNA fragmentation, chromatin condensation and changes in flow cytometry profiles detected during apoptosis. Exp. Cell Res. 217, 506–516.

    Article  PubMed  CAS  Google Scholar 

  40. Mitchell, M. S. (1988) Combining chemotherapy with biological response modifiers in treatment of cancer. J. Natl. Cancer Inst. 80, 1445–1450.

    Article  PubMed  CAS  Google Scholar 

  41. Morikawa, K., Hosokawa, M., Hamada, J., Xu, Z., and Kobayashi, H. (1986) Possible participation of tumoricidal macrophages in the therapeutic effect of blemycin on a transplantable rat fibrosarcoma. Cancer Res. 46, 684–688.

    PubMed  CAS  Google Scholar 

  42. Morikawa, K., Hosokawa, M., Hamada, J., Sugawara, M., and Kobayashi, H. (1985) Host-mediated therapeutic effects produced by appropriately timed administration of bleomycin on a rat fibrosarcoma. Cancer Res. 45, 1502–1506.

    PubMed  CAS  Google Scholar 

  43. Xu, Z., Hosokawa, M., Morikawa, K., Hatakeyama, M., and Kobayashi, H. (1988) Overcoming suppression of antitumor immune reactivity in tumor-bearing rats by treatment with bleomycin. Cancer Res. 48, 6658–6663.

    PubMed  CAS  Google Scholar 

  44. Abdul Hamied, T. A. and Turk, J. L. (1987) Enhancement of interleukin-2 release in rats by treatment with bleomycin and adriamycin in vivo. Cancer Immunol. Immunother. 25, 245–249.

    PubMed  CAS  Google Scholar 

  45. Heller, R., Jaroszeski, M. J., Perrott, R., Messina, J., and Gilbert, R. (1997) Effective treatment of B16 melanoma by direct delivery of bleomycin using electrochemotherapy. Melanoma Res. 7, 10–18.

    Article  PubMed  CAS  Google Scholar 

  46. Jaroszeski, M. J., Gilbert, R., and Heller, R. (1997) In vivo antitumor effects of electrochemotherapy in a hepatoma model. Biochim. Biophys. Acta 1334, 15–18.

    CAS  Google Scholar 

  47. Heller, R., Jaroszeski, M. J., Reintgen, D. S., Puleo, C. A., DeConti, R. C., Gilbert, R. A., and Glass, L. F. (1998) Treatment of cutaneous and subcutaneous tumors with electrochemotherapy using intralesional bleomycin. Cancer 83, 148–157.

    Article  PubMed  CAS  Google Scholar 

  48. Gately, D. F. and Howell, S. B. (1993) Cellular accumulation of the anticancer agent cisplatin: A review. Br. J. Cancer 67, 1171–1176.

    Article  PubMed  CAS  Google Scholar 

  49. Melvik, J. E., Pettersen, E. O., Gordon, P. B., and Seglen, P. O. (1986) Increase in cis-dichlorodiammineplatinum(II) cytotoxicity upon reversible electropermeabilization of the plasma membrane in cultured human NHIK 3025 cells. Eur. J. Cancer Clin. Oncol. 22, 1523–1530.

    Article  CAS  Google Scholar 

  50. Serša, G., Čemažar, M., and Miklavčič, D. (1995) Antitumor effectiveness of electrochemotherapy with cis-diamminedichloroplatinum(II) in mice. Cancer Res. 55, 3450–3455.

    PubMed  Google Scholar 

  51. Čemažar, M., Milačič, R., Miklavčič, D., Dolžan, V., and Serša, G. (1998) Intratumoral cisplatin administration in electrochemotherapy: antitumor effectiveness, sequence dependence and platinum content. Anti-Cancer Drugs 9, 525–530.

    Article  PubMed  Google Scholar 

  52. Čemažar, M., Miklavčič, D., Ščančar, J., Dolžan, V., Golouh, R., and Serša, G. (1999) Increased platinum accumulation in SA-1 tumour cells after in vivo electrochemotherapy with cisplatin. Br. J. Cancer 79, 1386–1391.

    Article  PubMed  Google Scholar 

  53. Ramirez, L. H., Orlowski, S., An, D. J., Bindoula, G., Dzodic, R., Ardouin, P., Bognel, C., Belehradek, J. Jr., Munck, J.-N., and Mir, L. M. (1998) Electrochemotherapy on liver tumors in rabbits. Br. J. Cancer 77, 2104–2111.

    Article  PubMed  CAS  Google Scholar 

  54. Serša, G., Beravs, K., Čemažar, M., Miklavčič, D., and Demšar, F. (1998) Contrast enhanced MRI assesment of tumor blood volume after application of electric pulses. Electr. Magnetobiol. 17, 297–304.

    Google Scholar 

  55. Serša, G., Čemažar, M., Parkins, C. S., and Chaplin, D. J. (1999) Tumour blood flow changes induced by application of electric pulses. Eur. J. Cancer 35, 672–677.

    Article  PubMed  Google Scholar 

  56. Serša, G., Miklavčič, D., Čemažar, M., Belehradek, J. Jr., Jarm, T., and Mir, L. M. (1997) Electrochemotherapy with CDDP on LPB sarcomas: Comparison of the anti-tumor effectiveness in immunocompetent and immunodeficient mice. Bioelectrochem. Bioenerg. 43, 279-283.

    Google Scholar 

  57. Mir, L. M., Orlowski, S., Poddevin, B., and Belehradek, J. Jr. (1992) Electrochemotherapy tumor treatment is improved by interleukin-2 stimulation of host’s defenses. Eur. Cytokine Netw. 3, 331–334.

    PubMed  CAS  Google Scholar 

  58. Serša, G., Čemažar, M., Menart, V., Gaberc-Porekar, V., and Miklavčič, D. (1997) Anti-tumor effectiveness of electrochemotherapy with bleomycin is increased by TNF-α on SA-1 tumors in mice. Cancer Lett. 116, 85–92.

    Article  PubMed  Google Scholar 

  59. Mir, L. M., Roth, C., Orlowski, S., Quintin-Colonna, F., Fradelizi, D., Belehradek, J. Jr., and Kourilsky, P. (1995) Systemic antitumor effects of electrochemotherapy combined with histoincompatible cells secreting interleukin-2. J. Immunother. 17, 30–38.

    Article  CAS  Google Scholar 

  60. Mir, L. M., Roth, C., Orlowski, S., Belehradek, J. Jr., Fradelizi, D., Paoletti, C., and Kourilsky, P. (1992) Potentiation of the antitumoral effect of electrochemotherapy by an immunotherapy with allogeneic cells producing interleukin-2. C. R. Acad. Sci. III 314, 539–544.

    PubMed  CAS  Google Scholar 

  61. Roth, C., Mir, L. M., Cressent, M., Quintin-Colonna, F., Ley, V., Fradelizi, D., and Kourilsky, P. (1992) Inhibition of tumor growth by histoincompatible cells expressing interleukin-2. Int. Immunol. 4, 1429–1436.

    Article  PubMed  CAS  Google Scholar 

  62. Orlowski, S., An, D., Belehradek, J. Jr., and Mir, L. M. (1998) Antimetastatic effects of electrochemotherapy and of histoincompatible interleukin-2-secreting cells in the murine Lewis lung tumor. Anticancer Drugs bd9, 551–556.

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2000 Humana Press Inc., Totowa, NJ

About this protocol

Cite this protocol

Mir, L.M., Orlowski, S. (2000). The Basis of Electrochemotherapy. In: Jaroszeski, M.J., Heller, R., Gilbert, R. (eds) Electrochemotherapy, Electrogenetherapy, and Transdermal Drug Delivery. Methods in Molecular Medicine, vol 37. Humana Press. https://doi.org/10.1385/1-59259-080-2:99

Download citation

  • DOI: https://doi.org/10.1385/1-59259-080-2:99

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-606-2

  • Online ISBN: 978-1-59259-080-3

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics