AMPK-mTOR-ULK1 Activation-Dependent Autophagy Compromises Oleanolic Acid-induced Cytotoxicity Inhuman Bladder Cancer Cells

Objective: To understand the protective autophagy process when the oleanolic acid induced apoptosis of human bladder cancer cells;and the effect of this progress that compromises the apoptosis of bladder cancer cell line;finally put forward the application of oleanolic acid combined with autophagy inhibitor for the treatment of bladder cancer can increase the therapeutic effect.Methods:Pick up two bladder cancer cell lines: T24 and EJ.T24 and EJ cells were treated with different concentrations of OA for 24 h,using CCK8 assay for testing cell viability;Using colony formation assay for testing cell proliferation.The cells were then stained with Annexin V/FITC and PI,and then flow cytometrically examined.Extraction of protein of two cell lines to test apoptosis related proteins BAX,Bcl-2,Caspase-3.We transfect with m Cherry-GFP-LC3 B,with or without oleanolic acid under the condition of 12 hours of training,to observe autophagy under the fluorescence microscope.Using western-blot to test LC3 B,p-AMPK,p-m TOR,p-ULK1 to determine the mechanism of autophagy.Using 3-MA and rapamycin to test whether autophagy can effect OA-induced apoptosis.Using dorsomorphin inhibit AMPK to show the different of protective autophagy upon OA-induced apotosis.Results: OA inhibited viability and proliferation of human bladder cancer cells.OA induced apoptosis in human bladder cancer cells.OA induced apoptosis in human bladder cancer cells.OA induced apoptosis in human bladder cancer cells.OA induced apoptosis in human bladder cancer cells.OA induced apoptosis in human bladder cancer cells.AMPK was implicated in protective autophagy upon OAinduced apoptosis in T24 cells.Conclution: Confirm the oleanolic acid giving rise to protective autophagy through the activation of the AMPK-m TOR-ULK1 axis in bladder cancer.OA in combination with autophagy inhibitors might be a novel alternative for the treatment of bladder cancer.