RÉSUMÉ
Objective To perform comparative proteomic analysis of human ovarian cancer cell lines for detecting platinum-resistance associated proteins.Methods The total proteins of two sensitive (SKOV3 and A2780)and four resistant(SKOV3/CDDP,SKOV3/CBP,A2780/CDDP and A2780/CBP) human ovarian cancer cell lines were separated by two-dimensional gel electrophoresis(2-DE).The differentially expressed proteins were analyzed using image analysis software,stained with Coomassie Brilliant Blue,then identified using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry(MALDI-TOF-MS)and database searching.The mRNA and protein levels of the differentially expressed protein which was most significant in all of the four resistant cell lines were validated by RT-PCR and western blotting,respectively.Results Five proteins were found to be significant in four cell lines. Annexin A3 and destrin were up-regulated and nicotinamide-adenine dinucleotide phosphate(NADP)- dependent isocitrate dehydrogenase 1 was down-regulated in all the four resistant samples.Glutathione transferase omega 1 had an increased expression in the other three resistant cell lines except for SKOV3/CBP in which its expression was not changed.However,cofilin 1 represented a different trend.In the two resistant sublines of SKOV3,eofilin 1 had a down-regulation,but it had an up-regulation in the cell lines induced from SKOV3.The expression of annexin A3 was up-regulated by 3-20 fold and the results of RT- PCR and western blotting showed complete consistency with that by 2-DE.Conclusions Proteomic techniques are useful to the identification of the resistance-associated proteins in ovarian cancer platinum- resistant cell lines and five candidates have been found.The five differential proteins might become hopeful candidate biomarkers for resistance.