RESEARCH ARTICLE A proteomic kinetic analysis of IGROV1 ovarian carcinoma cell line response to cisplatin treatment Karen Le Moguen 1 , Hubert Lincet 1 , Paulo Marcelo 2 , Edwige Lemoisson 1 , Natacha Heutte 1 , Marilyne Duval 1 , Laurent Poulain 1 , Joëlle Vinh 2 , Pascal Gauduchon 1 and Bruno Baudin 1, 3 1 Groupe Régional d’Etudes sur le Cancer – EA 1772 (Université de Caen-Basse Normandie), Unité Biologie et Thérapies Innovantes des Cancers Localement Agressifs, Centre de Lutte Contre le Cancer Franc ¸ ois Baclesse, Caen, France 2 Unité de Neurobiologie et Diversité Cellulaire, CNRS-UMR 7637, ESPCI, Paris, France 3 Service de Biochimie A, Hôpital Saint-Antoine, AP-HP, Paris, France Ovarian cancer is one of the leading causes of mortality by gynecological cancer. Despite good re- sponse to surgery and initial chemotherapy, essentially based on cisplatin (cis-diamino-dichloro- platinum(II) (CDDP)) compounds, frequent recurrences with chemoresistance acquisition are responsible for poor prognosis. Several mechanisms have been described as implicated in CDDP resistance, however they are not sufficient to exhaustively account for this resistance emergence. We applied a proteomic approach based on 2-DE coupled with MS (MALDI-TOF/TOF) to identify proteins associated with chemoresistance induced by CDDP. A kinetic analysis of IGROV1 cell behavior following treatment with CDDP and subsequent statistical analysis revealed time and/or concentration-dependent modifications in protein expression. We evidenced events such as decreased amino-acid and nucleotide synthesis potentially associated with cell cycle blockade, and variations that may be related to resistance acquisition, such as possible enhanced glycolysis and increased proliferating potential. Moreover, overexpressions of aldehyde dehydrogenase 1 and both cytokeratins 8 and 18 were consistent with our previous findings, demonstrating that expression of these proteins was increased in cisplatin-resistant IGROV1-R10 as compared to IGROV1 parental cells. Identification of such proteins could allow improved understanding of the mechanisms leading to cell death or survival and, thus, to the acquisition of chemoresistance. Received: March 7, 2007 Revised: July 4, 2007 Accepted: July 31, 2007 Keywords: Cisplatin / Drug resistance / Ovarian cancer / Tumor cell lines 4090 Proteomics 2007, 7, 4090–4101 1 Introduction Cisplatin (cis-diamino-dichloro-platinum(II) (CDDP)) is a platinum-based antitumor drug that forms intra- and inter- strand adducts with DNA. It is widely used in the treatment of solid tumors, in particular ovarian cancers. Despite good response to surgery and first-line chemotherapy based on platinum derivatives, frequent recurrences associated with chemoresistance acquisition are responsible for the poor prognosis of this cancer, with an overall 5 year survival of 30%. Thus, ovarian cancer remains the leading cause of mortality by gynecological cancer in Europe and the USA, affecting approximately 1 in 75 women. The response to CDDP is a complex and multifactorial process that leads to Correspondence: Dr. Karen Le Moguen, Groupe Régional d’Etudes sur le Cancer – EA 1772 (Université de Caen-Basse Nor- mandie), Unité Biologie et Thérapies Innovantes des Cancers Localement Agressifs, Centre de Lutte Contre le Cancer Franc ¸ ois Baclesse, Ave du Général Harris, 14076 Caen Cedex 05, France E-mail: k.le.moguen@baclesse.fr Fax: 133-2-31-455172 Abbreviations: ALDH, aldehyde dehydrogenase; ANX, annexin; CDDP, cis-diamino-dichloro-platinum(II) (cisplatin); CK, cytokera- tin; DAPI, 4 0 ,6-diamidino-2-phenylindole; hnRNP, heterogeneous nuclear ribonucleoprotein; IMP-dehydrogenase, inosine-5 0 - monophosphate dehydrogenase; 3-PGDH:, D-3-phosphoglyce- rate dehydrogenase; RUVBL1, RuvB like 1; SOD, superoxide dis- mutase; TCTP, translationally controlled tumor protein; TPI, trio- sephosphate isomerase; trpRS, tryptophanyl tRNA synthetase DOI 10.1002/pmic.200700231  2007 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim www.proteomics-journal.com