title: Autophagy regulates colistin-induced apoptosis in PC-12 cells.
abstract: Colistin is a cyclic cationic polypeptide antibiotic with activity against multidrug-resistant Gram-negative bacteria. Our recent study demonstrated that colistin induces apoptosis in primary chick cortex neurons and PC-12 cells. Although apoptosis and autophagy have different impacts on cell fate, there is a complex interaction between them. Autophagy plays an important role as a homeostasis regulator by removing excessive or unnecessary proteins and damaged organelles. The aim of the present study was to investigate the modulation of autophagy and apoptosis regulation in PC-12 cells in response to colistin treatment. PC-12 cells were exposed to colistin (125 to 250 μg/ml), and autophagy was detected by visualization of monodansylcadaverine (MDC)-labeled vacuoles, LC3 (microtubule-associated protein 1 light chain 3) immunofluorescence microscopic examination, and Western blotting. Apoptosis was measured by flow cytometry, Hoechst 33258 staining, and Western blotting. Autophagosomes were observed after treatment with colistin for 12 h, and the levels of LC3-II gene expression were determined; observation and protein levels both indicated that colistin induced a high level of autophagy. Colistin treatment also led to apoptosis in PC-12 cells, and the level of caspase-3 expression increased over the 24-h period. Pretreatment of cells with 3-methyladenine (3-MA) increased colistin toxicity in PC-12 cells remarkably. However, rapamycin treatment significantly increased the expression levels of LC3-II and beclin 1 and decreased the rate of apoptosis of PC-12 cells. Our results demonstrate that colistin induced autophagy and apoptosis in PC-12 cells and that the latter was affected by the regulation of autophagy. It is very likely that autophagy plays a protective role in the reduction of colistin-induced cytotoxicity in neurons.
authors: College of Veterinary Medicine, Northeast Agricultural University, Harbin, People's Republic of China Institute of Animal Husbandry and Veterinary Medicine, Liaoning Medical College, Jinzhou, Liaoning, People's Republic of China.
authors: Ling
authors: L
authors: Zhang
authors: College of Veterinary Medicine, Northeast Agricultural University, Harbin, People's Republic of China.
authors: Yonghao
authors: Y
authors: Zhao
authors: College of Veterinary Medicine, Northeast Agricultural University, Harbin, People's Republic of China.
authors: Wenjian
authors: W
authors: Ding
authors: College of Veterinary Medicine, Northeast Agricultural University, Harbin, People's Republic of China.
authors: Guozheng
authors: G
authors: Jiang
authors: College of Veterinary Medicine, Northeast Agricultural University, Harbin, People's Republic of China.
authors: Ziyin
authors: Z
authors: Lu
authors: Institute of Animal Husbandry and Veterinary Medicine, Liaoning Medical College, Jinzhou, Liaoning, People's Republic of China.
authors: Li
authors: L
authors: Li
authors: Institute of Animal Husbandry and Veterinary Medicine, Liaoning Medical College, Jinzhou, Liaoning, People's Republic of China.
authors: Jinli
authors: J
authors: Wang
authors: Drug Delivery, Disposition and Dynamics, Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, Victoria, Australia Colistin.Polymyxin@gmail.com lijichang@neau.edu.cn.
authors: Jian
authors: J
authors: Li
authors: College of Veterinary Medicine, Northeast Agricultural University, Harbin, People's Republic of China Colistin.Polymyxin@gmail.com lijichang@neau.edu.cn.
authors: Jichang
authors: J
authors: Li
mesh_terms: D000225:Adenine / Q000031:analogs & derivatives / Q000494:pharmacology; D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D051017:Apoptosis Regulatory Proteins / Q000378:metabolism; D001343:Autophagy / Q000502:physiology*; D000071186:Beclin-1; D053148:Caspase 3 / Q000378:metabolism; D003091:Colistin / Q000494:pharmacology*; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D016716:PC12 Cells; D051381:Rats; D014617:Vacuoles / Q000187:drug effects
chemical_list: D000225:Adenine; D000900:Anti-Bacterial Agents; D051017:Apoptosis Regulatory Proteins; D000071186:Beclin-1; D053148:Caspase 3; D003091:Colistin; D008869:Microtubule-Associated Proteins; C512829:Becn1 protein, rat; C526109:Anxa3 protein, rat; C025946:3-methyladenine
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