p53 Mediates Colistin-Induced Autophagy and Apoptosis in PC-12 Cells.
The mechanism of colistin-induced neurotoxicity is still unknown. Our recent study (L. Zhang, Y. H. Zhao, W. J. Ding, G. Z. Jiang, Z. Y. Lu, L. Li, J. L. Wang, J. Li, and J. C. Li, Antimicrob Agents Chemother 59:2189-2197, 2015, http://dx.doi.org/10.1128/AAC.04092-14; H. Jiang, J. C. Li, T. Zhou, C. H. Wang, H. Zhang, and H. Wang, Int J Mol Med 33:1298-1304, 2014, http://dx.doi.org/10.3892/ijmm.2014.1684) indicates that colistin induces autophagy and apoptosis in rat adrenal medulla PC-12 cells, and there is interplay between both cellular events. As an important cellular stress sensor, phosphoprotein p53 can trigger cell cycle arrest and apoptosis and regulate autophagy. The aim of the present study was to investigate the involvement of the p53 pathway in colistin-induced neurotoxicity in PC-12 cells. Specifically, cells were treated with colistin (125 μg/ml) in the absence and presence of a p53 inhibitor, pifithrin-α (PFT-α; 20 nM), for 12 h and 24 h, and the typical hallmarks of autophagy and apoptosis were examined by fluorescence/immunofluorescence microscopy and electron microscopy, real-time PCR, and Western blotting. The results indicate that colistin had a stimulatory effect on the expression levels of the target genes and proteins involved in autophagy and apoptosis, including LC3-II/I, p53, DRAM (damage-regulated autophagy modulator), PUMA (p53 upregulated modulator of apoptosis), Bax, p-AMPK (activated form of AMP-activated protein kinase), and caspase-3. In contrast, colistin appeared to have an inhibitory effect on the expression of p-mTOR (activated form of mammalian target of rapamycin), which is another target protein in autophagy. Importantly, analysis of the levels of p53 in the cells treated with colistin revealed an increase in nuclear p53 at 12 h and cytoplasmic p53 at 24 h. Pretreatment of colistin-treated cells with PFT-α inhibited autophagy and promoted colistin-induced apoptosis. This is the first study to demonstrate that colistin-induced autophagy and apoptosis are associated with the p53-mediated pathway.
College of Veterinary Medicine, Northeast Agricultural University, Harbin, People's Republic of China Institute of Animal Husbandry and Veterinary Medicine, Jinzhou Medical University, Liaoning, People's Republic of China.
Ling
L
Zhang
College of Veterinary Medicine, Northeast Agricultural University, Harbin, People's Republic of China.
Daoyuan
D
Xie
College of Veterinary Medicine, Northeast Agricultural University, Harbin, People's Republic of China.
Xueping
X
Chen
Drug Delivery, Disposition and Dynamics, Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, Victoria, Australia.
Maria L R
ML
Hughes
College of Veterinary Medicine, Northeast Agricultural University, Harbin, People's Republic of China.
Guozheng
G
Jiang
College of Veterinary Medicine, Northeast Agricultural University, Harbin, People's Republic of China.
Ziyin
Z
Lu
College of Veterinary Medicine, Northeast Agricultural University, Harbin, People's Republic of China.
Chunli
C
Xia
Institute of Animal Husbandry and Veterinary Medicine, Jinzhou Medical University, Liaoning, People's Republic of China.
Li
L
Li
Institute of Animal Husbandry and Veterinary Medicine, Jinzhou Medical University, Liaoning, People's Republic of China.
Jinli
J
Wang
Institute of Animal Husbandry and Veterinary Medicine, Jinzhou Medical University, Liaoning, People's Republic of China.
Wei
W
Xu
Institute of Animal Husbandry and Veterinary Medicine, Jinzhou Medical University, Liaoning, People's Republic of China.
Yuan
Y
Sun
College of Veterinary Medicine, Northeast Agricultural University, Harbin, People's Republic of China.
Rui
R
Li
College of Veterinary Medicine, Northeast Agricultural University, Harbin, People's Republic of China.
Rui
R
Wang
College of Veterinary Medicine, Northeast Agricultural University, Harbin, People's Republic of China.
Feng
F
Qian
Drug Delivery, Disposition and Dynamics, Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, Victoria, Australia lijichang@neau.edu.cn Colistin.Polymyxin@gmail.com.
Jian
J
Li
College of Veterinary Medicine, Northeast Agricultural University, Harbin, People's Republic of China lijichang@neau.edu.cn Colistin.Polymyxin@gmail.com.
Jichang
J
Li
D055372:AMP-Activated Protein Kinases / Q000235:genetics / Q000378:metabolism; D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D051017:Apoptosis Regulatory Proteins / Q000819:agonists / Q000235:genetics / Q000378:metabolism; D001343:Autophagy / Q000187:drug effects*; D052160:Benzothiazoles / Q000494:pharmacology; D053148:Caspase 3 / Q000235:genetics / Q000378:metabolism; D002454:Cell Differentiation; D003091:Colistin / Q000494:pharmacology*; D005786:Gene Expression Regulation; D008565:Membrane Proteins / Q000819:agonists / Q000235:genetics / Q000378:metabolism; D008869:Microtubule-Associated Proteins / Q000819:agonists / Q000235:genetics / Q000378:metabolism; D009474:Neurons / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D016716:PC12 Cells; D051381:Rats; D015398:Signal Transduction; D058570:TOR Serine-Threonine Kinases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D014050:Toluene / Q000031:analogs & derivatives / Q000494:pharmacology; D016159:Tumor Suppressor Protein p53 / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D051028:bcl-2-Associated X Protein / Q000819:agonists / Q000235:genetics / Q000378:metabolism
D055372:AMP-Activated Protein Kinases; D000900:Anti-Bacterial Agents; D051017:Apoptosis Regulatory Proteins; D052160:Benzothiazoles; D053148:Caspase 3; D003091:Colistin; D008565:Membrane Proteins; D008869:Microtubule-Associated Proteins; D058570:TOR Serine-Threonine Kinases; D014050:Toluene; D016159:Tumor Suppressor Protein p53; D051028:bcl-2-Associated X Protein; C494828:Bax protein, rat; C533952:Bbc3 protein, rat; C000610489:DRAM1 protein, rat; C526109:Anxa3 protein, rat; C121565:pifithrin; C546845:mTOR protein, rat; C505410:Casp3 protein, rat