Cell adhesion to fibronectin induces mitomycin C resistance in bladder cancer cells.

OBJECTIVE: To investigate whether cell adhesion to fibronectin induces drug resistance in human bladder cancer cells, and to study the survival signalling pathway in cell adhesion to fibronectin-mediated chemotherapy resistance in vitro. MATERIALS AND METHODS: T24 cells (human bladder cancer cell lines) were pre-coated with fibronectin, and treated with mitomycin C (MMC) and the specific phosphoinositide-3 kinase (PI3-K) inhibitor LY294002. The apoptosis and cell cycles were analysed. The activity of the caspase-8, -9 and apoptosis-inducing factor (AIF) apoptosis pathways were assessed using colorimetric assay, immunofluorescence, Western blot and flow cytometry. The expression of glycogen synthase kinase-3beta (GSK-3beta) and cyclin D1, as the key regulator of G1/S phase transition, were determined by Western blot. The expression of PI3-K, Akt, phospho-Akt and beta1-integrin were also examined by Western blot. RESULTS: Apoptosis induced by MMC was significantly resisted by fibronectin adhesion in T24 cells, and this effect was through inhibition of the caspase-9 and AIF apoptosis pathways, but not the caspase-8 pathway. Fibronectin antagonized MMC-induced G0/G1-phase arrest by inactivating GSK-3beta to stabilize cyclin D1 expression in T24 cells. Furthermore, fibronectin-mediated protection of T24 cells was dependent on the activity of the PI3-K/Akt signalling pathway, and the protection could be abolished by the PI3-K inhibitor LY294002. CONCLUSIONS: Fibronectin-mediated PI3-K/Akt activation protects T24 cells from MMC-induced cell death through inhibition of both caspase-9 and AIF-mediated apoptosis and GSK-3beta/cyclin D1 involved G0/G1-phase arrest.