Original article Combination of therapy with 5-fluorouracil and cisplatin with electroporation in human ovarian carcinoma model in vitro Jolanta Saczko a , Iwona Kamin ´ ska b , Malgorzata Kotulska c , Julia Bar b , Anna Choroman ´ ska a , Nina Rembiałkowska a , Katarzyna Biez ˙ un ´ ska-Kusiak a , Joanna Rossowska d , Danuta Nowakowska e , Julita Kulbacka a, * a Department of Medical Biochemistry, Wroclaw Medical University, 10, street Chalubinskiego, 50-368 Wroclaw, Poland b Department of Pathomorphology and Clinical Cytology, Wroclaw Medical University, 213, street Borowska, 50-556 Wroclaw, Poland c Institute of Biomedical Engineering and Instrumentation, Wroclaw University of Technology, Wybrzeze Wyspianskiego 27, 50-370 Wroclaw, Poland d Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, 12, street Rudolf Weigl, 53-114 Wroclaw, Poland e Department of Dental Materials, Wroclaw Medical University, 26, street Krakowska, 50-425 Wroclaw, Poland 1. Introduction Ovarian cancer is the most lethal type among the gynecological cancers. Despite recent developments in aggressive surgery and chemotherapy (CT), the prognosis remains poor because ovarian cancers are often not diagnosed until advanced stages. More- over, the cancer cells often have primary or secondary drug resistance [1]. The successful treatment of cancer is dependent on effectiveness of cytotoxic anticancer therapies, either alone or in combination with other treatments. Therefore, new diagnostic and therapeutic techniques, with significantly improved efficacy, are required. Electrochemotherapy (ECT) is a relatively new method of anticancer therapy [2]. This method offers an alternative treatment to conventional therapies, and it is becoming increasingly accepted as a therapeutic modality in oncology [3–7]. This technique is a combination of conventional chemotherapy with cell membrane electroporation (EP), increasing drug transport into cells. In electrochemotherapy, cells exposed to EP may display 300– 700-fold increase of intracellular drug concentration, as in case of bleomycin – the drug applied the most frequently in the electrochemotherapy, which is normally non-permeant through plasma membrane [8,9]. In ECT, a weakly permeant drug is generally absorbed by cancer cells and shows the effect only in the cancer cells, significantly reducing side effects of the standard systemic chemotherapy. An important contribution to the therapy efficiency comes from the vascular lock in ECT, which keeps elevated drug concentrations in the vicinity of the cancer for several hours [10]. The appropriate therapy conditions could limit necessity of surgical intervention, and give better prognoses in treatment of the tumors. In contrast to chemotherapy and Biomedicine & Pharmacotherapy 68 (2014) 573–580 A R T I C L E I N F O Article history: Received 8 April 2014 Accepted 31 May 2014 Keywords: Electropermeabilization Electrochemotherapy Ovarian carcinoma Human fibroblasts 5-fluorouracil Cisplatin A B S T R A C T High electric field, applied to plasma membrane, affects organization of the lipid molecules, generating transient hydrophilic electropores. The application of the cell membrane electroporation in combination with cytotoxic drugs could increase the drug transport into cells. This approach is known as electrochemotherapy (ECT). Our work shows new data concerning the influence of electrochemical reaction with cisplatin or with 5-fluorouracil (5-FU) on cancer ovarian cells resistant to standard therapy with cisplatin, in comparison to ECT effect on human primary fibroblasts. We investigated the effect of electroporation and electrochemotherapy with 5-FU and cisplatin on human ovarian clear-cell carcinoma cell line (OvBH-1) and epithelial ovarian carcinoma cell line (SKOV-3) – both resistant to cisplatin typically used in ovarian cancers. As control cells, human gingival fibroblasts (HGF’s) from primary culture were used. Electropermeabilization efficiency was determined by FACS analysis with iodide propidium. Efficiency of electrochemotherapy was evaluated with viability assay. The cytotoxic effect was dependent on the electroporation parameters and on drug concentration. Electroporation alone only insignificantly decreased cells proliferation in OvBH-1 line; SKOV-3 line was more sensitive to the electrical field. Electrochemotherapy with cisplatin and 5-FU showed promising effects on both ovarian cell lines with recovery of normal cells revealed after 72 hours. ß 2014 Elsevier Masson SAS. All rights reserved. * Corresponding author. Tel.: +48 71 7821387. E-mail address: Julita.Kulbacka@umed.wroc.pl (J. Kulbacka). Available online at ScienceDirect www.sciencedirect.com http://dx.doi.org/10.1016/j.biopha.2014.05.005 0753-3322/ß 2014 Elsevier Masson SAS. All rights reserved.