title: Proteomic (antibody microarray) exploration of the molecular mechanism             of action of the specific COX-2 inhibitor DuP 697.
abstract: We have previously shown that specific COX-2 inhibitors, including DuP 697, have anti-proliferative effects on mesothelioma cells and potentiate the cytotoxicity of pemetrexed. Here, we used a novel proteomic approach to explore the mechanism of action of this agent. COX-2-positive cell lines MSTO-211H (mesothelioma) and A549 (lung cancer) were exposed to DuP 697 for 72 h. Drug carrier only was added to control cells. Extracted proteins from treated and control cells were analysed using a comparative proteomic platform. Differentially expressed proteins, identified by the Panorama Xpress Profiler725 antibody microarray were submitted to Ingenuity Pathway Analysis. A total of 32 unique differentially expressed proteins were identified with a significant (>1.8-fold) difference in expression between treated and untreated cells in at least one cell line. Five molecules, BCL2L1 (Bcl-xL), BID, CHUK (IKK), FASLG and RAF1, were mapped to the Apoptosis Signaling pathway following Ingenuity Pathway Analysis. BCL2L1 (Bcl-xL) and BID were analysed using immuno-blotting and differential expression was confirmed. Proteomic (antibody microarray) analysis suggests that the mechanism of action of DuP 697 may be exerted via the induction of apoptosis. The antibody microarray platform can be utilised to explore the molecular mechanism of action of novel anticancer agents.
authors: Cancer Biology Proteomics Group, Postgraduate Medical Institute                     of the University of Hull, Hull, UK.
authors: Vijay
authors: V
authors: Agarwal
authors: Victoria C
authors: VC
authors: Hodgkinson
authors: Gina L
authors: GL
authors: Eagle
authors: Lucy
authors: L
authors: Scaife
authors: Michael J
authors: MJ
authors: Lind
authors: Lynn
authors: L
authors: Cawkwell
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects*; D051023:BH3 Interacting Domain Death Agonist Protein / Q000032:analysis; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D051546:Cyclooxygenase 2 / Q000378:metabolism; D052246:Cyclooxygenase 2 Inhibitors / Q000494:pharmacology*; D005971:Glutamates / Q000494:pharmacology; D006147:Guanine / Q000031:analogs & derivatives / Q000494:pharmacology; D006801:Humans; D008175:Lung Neoplasms / Q000188:drug therapy* / Q000378:metabolism; D008654:Mesothelioma / Q000188:drug therapy* / Q000378:metabolism; D000068437:Pemetrexed; D040081:Protein Array Analysis; D040901:Proteomics; D013876:Thiophenes / Q000494:pharmacology*; D051020:bcl-X Protein / Q000032:analysis
chemical_list: D000970:Antineoplastic Agents; C494811:BCL2L1 protein, human; D051023:BH3 Interacting Domain Death Agonist Protein; C494814:BID protein, human; D052246:Cyclooxygenase 2 Inhibitors; D005971:Glutamates; D013876:Thiophenes; D051020:bcl-X Protein; D000068437:Pemetrexed; D006147:Guanine; C064874:DuP 697; D051546:Cyclooxygenase 2; C496540:PTGS2 protein, human