Enhanced cytotoxicity of bleomycin and cisplatin after electroporation in murine colorectal carcinoma cells
Background. Electrochemotherapy is a local treatment combining application of electric pulses and chemotherapy. Two chemotherapeutic drugs, bleomycin and cisplatin, have proved to be effective in electrochemotherapy. The effectiveness of electrochemotherapy was demonstrated in the treatment of various cutaneous and subcutaneous tumours in cancer patients. Only a few preclinical studies were performed in colorectal carcinoma, mostly using bleomycin. Our aim was to evaluate the sensitivity of the murine colorectal carcinoma cell line CMT-93 to electrochemotherapy with bleomycin and cisplatin for potential use in preclinical and clinical studies.
Methods. CMT-93 cells were exposed to either the chemotherapeutic drug alone or electrochemotherapy. A clonogenic assay was used to determine cell survival after treatment. Apoptosis was measured by caspase-3/7 activity, necrosis by changes in cell morphology and cell viability by the MTS assay 16 hours after electrochemotherapy.
Results. Cells treated with electrochemotherapy were 500-fold more sensitive to bleomycin and 2.8-fold more sensitive to cisplatin compared to cells treated with the drugs alone. At the highest concentrations, a significant reduction in cell viability, increase in caspase-3/7 activity and necrotic cells were observed after electrochemotherapy.
Conclusions. Exposure of cells to electric pulses enhanced cytotoxicity of both bleomycin and cisplatin. Reduced cell viability was due to apoptotic and necrotic cell death. Furthermore, electrochemotherapy with bleomycin was more cytotoxic than electrochemotherapy with cisplatin in this colorectal carcinoma cell line.
If the inline PDF is not rendering correctly, you can download the PDF file here.
Sersa G Cemazar M Miklavcic D Rudolf Z. Electrochemotherapy of tumours. Radiol Oncol 2006; 40: 163-74.
Teissie J Escoffre JM Rols MP Golzio M. Time dependence of electric field effects on cell membranes. A review for a critical selection of pulse duration for therapeutical applications. Radiol Oncol 2008; 42: 196-206.
Cemazar M Sersa G Miklavcic D. Electrochemotherapy with cisplatin in the treatment of tumour cells resistant to cisplatin. Anticancer Res 1998; 18: 4463-6.
Orlowski S Belehradek J Jr Paoletti Mir LM. Transient electropermeabilization of cells in culture. Increase of the cytotoxicity of anticancer drugs. Biochem Pharmacol 1988; 37: 4727-33.
Jaroszeski MJ Dang V Pottinger C Hickey J Gilbert R Heller R. Toxicity of anticancer agents mediated by electroporation in vitro. Anticancer Drugs 2000; 11: 201-8.
Cemazar M Miklavcic D Mir LM Belehradek J Jr Bonnay M Fourcault D et al. Electrochemotherapy of tumours resistant to cisplatin: a study in a murine tumour model. Eur J Cancer 2001; 37: 1166-72.
Sersa G. The state-of-the-art of electrochemotherapy before the ESOPE study; advantages and clinical uses. EJC Suppl 2006; 4: 52-9.
Marty M Sersa G Garbay JR Gehl J Collins CG Snoj M et al. Electrochemotherapy - An easy highly effective and safe treatment of cutaneous and subcutaneous metastases: Results of ESOPE (European Standard Operating Procedures of Electrochemotherapy) study. EJC Suppl 2006; 4: 3-13.
Kuriyama S Matsumoto M Mitoro A Tsujinoue H Nakatani T Fukui H et al. Electrochemotherapy for colorectal cancer with commonly used chemotherapeutic agents in a mouse model. Dig Dis Sci 2000; 45: 1568-77.
Kuriyama S Tsujinoue H Toyokawa Y Mitoro A Nakatani T Yoshiji H et al. A potential approach for electrochemotherapy against colorectal carcinoma using a clinically available alternating current system with bipolar snare in a mouse model. Scand J Gastroenterol 2001; 36: 297-302.
Cemazar M Miklavcic D Sersa G. Intrinsic sensitivity of tumour cells to bleomycin as an indicator of tumour response to electrochemotherapy. Jpn J Cancer Res 1998; 45: 328-33.
Kranjc S Cemazar M Grosel A Pipan Z Sersa G. Effect of electroporation on radiosensitization with cisplatin in two cell lines with different chemo- and radiosensitivity. Radiol Oncol 2003; 37: 101-7.
Kambe M Arita D Kikuchi H Funato T Tezuka F Gamo M et al. Enhancement of the efficacy of anticancer drugs with electroporation: successful electrochemotherapy against gastric cancer cell lines in vivo and in vitro. Int J Clin Oncol 1997; 2: 111-7.
Sersa G Miklavcic D Cemazar M Rudolf Z Pucihar G Snoj M. Electrochemotherapy in treatment of tumours. EJSO 2008; 34: 232-40.
Fuertes MA Castilla J Alonso C Perez JM. Cisplatin biochemical mechanism of action: from cytotoxicity to induction of cell death through interconnections between apoptotic and necrotic pathways. Curr Med Chem 2003; 10: 257-66.
Cepeda V Fuertes M Castilla J Alonso C Quevedo C Perez JM. Biochemical mechanisms of cisplatin cytotoxicity. Anticancer Agents Med Chem 2007; 7: 3-18.
Tounekti O Kenani A Foray N Orlowski S Mir LM. The ratio of single- to double-strand DNA breaks and their absolute values determine cell death pathway. Brit J Cancer 2001; 84: 1272-9.
Chen J Stubbe J. Bleomycins: towards better therapeutics. Nat Rev Cancer 2005; 5: 102-12.
Siddik ZH. Cisplatin: mode of cytotoxic action and molecular basis of resistance. Oncogene 2003; 22: 7265-79.
Tounekti O Pron G Belehradek J Mir LM. Bleomycin an apoptosis-mimetic drug that induces two types of cell death depending on the number of molecules internalized. Cancer Res 1993; 53: 5462-9.
Gonzales VM Fuertes MA Alonso C Perez JM. Is cisplatin-induced cell death always produced by apoptosis? Mol Pharmacol 2001; 59: 657-63.
Wang D Lippard SJ. Cellular processing of platinum anticancer drugs. Nat Rev Drug Discov 2005; 4: 307-20.
Mekid H Tounekti O Spatz A Cemazar M El Kebir FZ Mir LM. In vivo evolution of tumour cells after the generation of double-strand DNA breaks. Brit J Cancer 2003; 88: 1763-71.
Vakifahmetoglu H Olsson M Tamm C Heidari N Orrenius S Zhivotovsky B. DNA damage induces two distinct modes of cell death in ovarian carcinomas. Cell Death Differ 2008; 15: 555-66.
Taylor RC Cullen SP Martin SJ. Apoptosis: controlled demolition at the cellular level. Nat Rev Cancer 2008; 9: 231-41.
Sadadcharam M Soden DM O'Sullivan GC. Electrochemotherapy: An emerging cancer treatment. Int J Hyperthermia 2008; 24: 263-73.