Proteomic Analysis of Rat PC12 Cells Exposed to Cyclosporin A
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Cyclosporin A (CsA) has been used clinically as animmunosuppressive drug to prevent organ transplantrejection and in basic research as a mitochondrialpermeability blocker. It has been reported that CsA has aprotective role in severed neurons and a neurotrophic effectin neuronal cells. However, the molecular mechanismsunderlying the stimulation of neuronal cell proliferation byCsA have not yet been elucidated. In our current study, weinvestigated CsA responsive proteins in PC12 cells using asystematic proteomic approach. The viability of these cellsfollowing CsA treatment increased in a dose- and timedependentmanner. Proteins in the CsA-treated PC12 cellswere profiled by two-dimensional gel electrophoresis (2-DE)and identified by matrix-assisted laser desorption ionizationtime-of flight (MALDI-TOF) and electrospray ionizationquadupole time-of-flight mass spectrometries (EIQ-TOFMS). This differential expression analysis showedsignificant changes for 10 proteins (6 up-regulated and 4down-regulated) upon CsA treatment that were related tocell proliferation, metabolism and the stress response. Theseproteomics data further our understanding of theproliferation mechanisms of PC12 cells exposed to CsA anddemonstrate that our methodology has potential to furtherelucidate the mechanisms and pathways involved.
Abstract
Cyclosporin A (CsA) has been used clinically as animmunosuppressive drug to prevent organ transplantrejection and in basic research as a mitochondrialpermeability blocker. It has been reported that CsA has aprotective role in severed neurons and a neurotrophic effectin neuronal cells. However, the molecular mechanismsunderlying the stimulation of neuronal cell proliferation byCsA have not yet been elucidated. In our current study, weinvestigated CsA responsive proteins in PC12 cells using asystematic proteomic approach. The viability of these cellsfollowing CsA treatment increased in a dose- and timedependentmanner. Proteins in the CsA-treated PC12 cellswere profiled by two-dimensional gel electrophoresis (2-DE)and identified by matrix-assisted laser desorption ionizationtime-of flight (MALDI-TOF) and electrospray ionizationquadupole time-of-flight mass spectrometries (EIQ-TOFMS). This differential expression analysis showedsignificant changes for 10 proteins (6 up-regulated and 4down-regulated) upon CsA treatment that were related tocell proliferation, metabolism and the stress response. Theseproteomics data further our understanding of theproliferation mechanisms of PC12 cells exposed to CsA anddemonstrate that our methodology has potential to furtherelucidate the mechanisms and pathways involved.
참고문헌 (33)
- Andre N, "Molecular effects of cyclosporin and oncogenesis: a new model", Med Hypotheses, 63, pp.647~652 (2004)
- Arcuri F, "Translationally controlled tumor protein (TCTP) in the human prostate and prostate cancer cells: expression, distribution, and calcium binding activity", Prostate, 60, pp.130~140 (2004)
- Bernardi P, "Modulation of the mitochondrial permeability transition pore. Effect of protons and divalent cations", J Biol Chem, 267, pp.2934~2939 (1994)
- Brockstedt E, "Identification of apoptosis-associated proteins in a human Burkitt lymphoma cell line. Cleavage of heterogeneous nuclear ribonucleoprotein A1 by caspase 3", J Biol Chem, 273, pp.28057~28064 (1998)
- Coley C, "Effect of cyclosporin-A on human gingival fibroblasts in vitro", J Dent Res, 65, pp.353~358 (1986)
- Faulds D, "A review of its pharmacodynamic and pharmacokinetic properties, and therapeutic use in immunoregulatory disorders", Drugs, 45, pp.953~1040 (1993)
- Folbergrova J, "Changes in the bioenergetic state of rat hippocampus during 2.5min of ischemia, and prevention of cell damage by cyclosporin A in hyperglycemic subjects", Exp Brain Res, 114, pp.44~50 (1997)
- Grabowski DT, "Drosophila AP3, a presumptive DNA repair protein, is homologous to human ribosomal associated protein P0", Nucleic Acids Res, 19, pp.4297~ (1991)
- Hartl FU, "Molecular chaperones in cellular protein folding", Nature, 381, pp.571~579 (1996)
- Hellman U, "Improvement of an "In-Gel" disgestion procedure for the micropreparation of internal protein fragments for amino acid sequencing", Anal Biochem, 224, pp.451~455 (1995)
- Hoffmann B, "Peroxiredoxins", Biol Chem, 383, pp.347~364 (2002)
- Kang SW, "Mammalian peroxiredoxin isoforms can reduce hydrogen peroxide generated in response to growth factors and tumor necrosis factor-α", J Biol Chem, 273, pp.6297~6302 (1998)
- Leprince P, "Modulation of proteolytic activity during neuritogenesis in the PC12 nerve cell: differential control of plasminogen activator and plasminogen activator inhibitor activities by nerve growth factor and dibutyryl-cyclic AMP", J Neurochem, 57, pp.665~674 (1991)
- Li F, "Characterization of fortilin, a novel antiapoptotic protein", J Biol Chem, 276, pp.47542~57549 (2001)
- Lyons WE, "Immunosuppressant FK506 promotes neurite outgrowth in cultures of PC12 cells and sensory ganglia", Proc Natl Acad Sci USA, 91, pp.3191~3195 (1994)
- Mascarell L, "Characterization of a gene encoding two isoforms of a mitochondrial protein up-regulated by cyclosporin A in activated T cells", J Biol Chem, 279, pp.10556~10563 (2004)
- Miyata K, "Involvement of the brainderived neurotrophic factor", TrkB pathway in neuroprotective effect of cyclosporin A in forebrain ischemia, 2001, pp.105, 571~578
- Moore MS, "Purification of a Ran-interacting protein that is required for protein import into the nucleus", Proc Natl Acad Sci USA, 91, pp.10212~10216 (1994)
- Noga M, "Gene expressions of ubiquitin and hsp70 following focal ischemia in rat brain", Neuroreport, 24, pp.1239~1241 (1997)
- 박지일, "Involvement of Antiapoptotic Signals in Rat PC12 Cells Proliferation by Cyclosporin A Treatment", International Journal of Oral Biology, 32(2), pp.51~57 (2007), 대한구강생물학회
- Prosperi MT, "The pag gene product, a physiological inhibitor of c-abl tyrosine kinase, is overexpressed in cells entering S phase and by contact with agents inducing oxidative stress", FEBS Lett, 423, pp.39~44 (1998)
- Quimby BB, "Functional analysis of the hydrophobic patch on nuclear transport factor 2 involved in interactions with the nuclear pore in vivo", J Biol Chem, 276, pp.38820~38829 (2001)
- Roobol A, "Cytoplasmic chaperonin complexes enter neurites developing in vitro and differ in subunit composition within single cells", J Cell Sci, 108, pp.1477~1488 (1995)
- Sanchez JC, "Translationally controlled tumor protein: A protein identified in several nontumoral cells including erythrocytes", Electrophoresis, 18, pp.150~155 (1997)
- Sandoval JA, "Novel peptides secreted from human neuroblastoma: useful clinical tools?", J Pediatr Surg, 41, pp.45~51 (2006)
- Scheler C, "Peptide mass fingerprint sequence coverage from differently stained proteins on two-dimensional electrophoresis patterns by matrix assisted laser desorption", ionization-mass spectrometry (MALDI-MS), 1998, pp.19, 918~927
- Shiga Y, "Cyclosporin Aprotects against ischemia-reperfusion injury in the brain", Brain Res, 595, pp.145~148 (1992)
- Takahashi T, "Cyclosporin a promotes hair epithelial cell proliferation and modulates protein kinase C expression and translocation in hair epithelial cells", J Invest Dermatol, 117, pp.605~611 (2001)
- Tanaka K, "Immunophilin ligands: from immunosuppressants to neuroprotective drugs", Nihon Shinkei Seishin Yakurigaku Zasshi, 24, pp.71~74 (2004)
- Thomas G, "Translational control of mRNA expression during the early mitogenic response in Swiss mouse 3T3 cells: Identification of specific proteins", J Cell Biol, 103, pp.2137~2144 (1986)
- Uchino H, "A dramatically ameliorates CA1 hippocampal damage following transient forebrain ischemia in the rat", Acta Physiol Scand, 155, pp.469~471 (1995)
- Yenofsky R, "Regulation of mRNA utilization in mouse erythroleukemia cells induced to differentiate by exposure to dimethyl sulfoxide", Mol Cell Biol, 3, pp.1197~1203 (1983)
- Zhang P, "Thioredoxin peroxidase is a novel inhibitor of apoptosis with a mechanism distinct from that of Bcl-2", J Biol Chem, 272, pp.30615~30618 (1997)