Inhibition of Mitochondria- and Endoplasmic Reticulum Stress-Mediated Autophagy Augments Temozolomide-Induced Apoptosis in Glioma Cells

• Published in: PloS one Vol. 7; no. 6; p. e38706
• Main Authors: Lin, Chien-Ju, Lee, Chin-Cheng, Shih, Yung-Luen, Lin, Chien-Huang, Wang, Sheng-Hao, Chen, Thay-Hsiung, Shih, Chwen-Ming
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 22.06.2012; Public Library of Science (PLoS)
• Subjects: Alkylation; Apoptosis; Apoptosis - drug effects; Autophagy; Autophagy - drug effects; Biochemistry; Biology; Brain; Brain cancer; Brain tumors; c-Jun protein; Calcium signalling; Cancer therapies; Caspase; Caspase-12; Cell death; Cell Line, Tumor; Cell survival; Cell Survival - drug effects; Cellular proteins; Chemotherapy; CHOP protein; Cyclosporins; Cytotoxicity; Dacarbazine - analogs & derivatives; Dacarbazine - pharmacology; Damage; Depolarization; DNA repair; Drug dosages; Electron transport; Electron transport chain; Endoplasmic reticulum; Endoplasmic Reticulum Stress - physiology; Explosives; Extracellular signal-regulated kinase; Flow cytometry; Glioma; Glioma - metabolism; Glioma cells; Gliomas; Homeostasis; Hospitals; Humans; Immunoblotting; Inhibition; Inhibitors; JNK protein; Kinases; Laboratories; Medicine; Melanoma; Membrane permeability; Membrane Potential, Mitochondrial - drug effects; Microscopy; Mitochondria; Mitochondria - drug effects; Mitochondria - metabolism; Mitochondrial permeability transition pore; MPTP; Oligomycin; Organelles; Oxidative stress; Oxygen; Permeability; Phagocytosis; Protein synthesis; Proteins; Reactive oxygen species; Reactive Oxygen Species - metabolism; Rotenone; Smooth muscle; Sodium; Stains & staining; Stem cells; Stresses; Studies; Temozolomide; Toxicity; Transcription factors; Taiwan
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0038706

Autophagy is a crucial process for cells to maintain homeostasis and survival through degradation of cellular proteins and organelles, including mitochondria and endoplasmic reticula (ER). We previously demonstrated that temozolomide (TMZ), an alkylating agent for brain tumor chemotherapy, induced reactive oxygen species (ROS)/extracellular signal-regulated kinase (ERK)-mediated autophagy to protect glioma cells from apoptosis. In this study, we investigated the role of mitochondrial damage and ER stress in TMZ-induced cytotoxicity. Mitochondrial depolarization and mitochondrial permeability transition pore (MPTP) opening were observed as a prelude to TMZ-induced autophagy, and these were followed by the loss of mitochondrial mass. Electron transport chain (ETC) inhibitors, such as rotenone (a complex I inhibitor), sodium azide (a complex IV inhibitor), and oligomycin (a complex V inhibitor), or the MPTP inhibitor, cyclosporine A, decreased mitochondrial damage-mediated autophagy, and therefore increased TMZ-induced apoptosis. TMZ treatment triggered ER stress with increased expression of GADD153 and GRP78 proteins, and deceased pro-caspase 12 protein. ER stress consequently induced autophagy through c-Jun N-terminal kinases (JNK) and Ca(2+) signaling pathways. Combination of TMZ with 4-phenylbutyrate (4-PBA), an ER stress inhibitor, augmented TMZ-induced cytotoxicity by inhibiting autophagy. Taken together, our data indicate that TMZ induced autophagy through mitochondrial damage- and ER stress-dependent mechanisms to protect glioma cells. This study provides evidence that agents targeting mitochondria or ER may be potential anticancer strategies.