Although growth free overnight delivery factor receptors such as EGFR and PDGFR can often activate the ERK pathway, and ligands of these receptors are present in Inhibitors,Modulators,Libraries OC ascites, we do not believe that the ascites mediated upregulation of Mcl 1 is dependent on these receptors because we previously shown Inhibitors,Modulators,Libraries that the inhib ition of EGFR and PDGFR does not alter the prosurvival activity of ascites. Our findings suggest that OC ascites activate multiple signaling pathways to inhibit TRAIL induced apoptosis and each pathway may contribute to a different level to ascites mediated protection from TRAIL depending, at least in part, on the cell context. Although the signifi cance of these in vitro observations in regard to the clinic has yet to be determined, we propose that ascites, by activating different survival pathways in tumor cells, contribute to the persistence of tumor cells during treat ment and the occurrence of resistance.

This has implica tion from a therapeutic standpoint. Targeting the tumor environment could be an important strategy to sensitize OC cells to chemotherapy. Materials and Inhibitors,Modulators,Libraries methods Cell culture and reagents The human OC cell lines CaOV3 and OVCAR3 were obtained from the American Type Culture Collection Inhibitors,Modulators,Libraries and maintained in a humidified 5% CO2 incubator at 37 C. Cells were passaged twice weekly. OVCAR3 cells were maintained in RPMI 1640 supplemented with 20% FBS, insu lin, glutamine Inhibitors,Modulators,Libraries and antibiotics. CaOV3 cells were cultured in DMEM/F12 supplemented with 10% FBS, 2 mM glutamine and antibiotics. TRAIL was purchased from PeproTech.

Acellular ascites fractions were obtained at the time of initial cytoreductive surgery from www.selleckchem.com/products/VX-770.html women with advanced serous ovarian carcinomas. Samples were supplied by the Banque d��chantillons biolo giques et de donn��es de Sherbrooke as part of the Banque de tissus et de donn��es du R��seau de Recherche en Cancer des Fonds de Recherche en Sant�� du Qu��bec affiliated to the Canadian Tumor Repository Network. HRP conjugated anti mouse and rabbit antibodies, Akt, Bcl XL, Elk 1, phos pho ERK1/2, Mcl 1, FAK, phospho FAK and phospho Elk 1 antibodies were purchased from Cell Signaling. Antibodies for phospho Akt were from Life Technologies. Bcl 2 anti body was purchased from Dako. ERK antibody was from Santa Cruz Biotech. PI3K inhibitor LY294002 and MEK inhibitor U0126 were purchased from EMD. Tubulin antibody, actinomycin D and propidium iodide were purchased from Sigma Aldrich. Actinomycin D was dissolved in dimethyl sulfoxide at a concentration of 10 mM and stored at ?20 C.