Abstract
Apoptosis and autophagy are closely interconnected types of programmed cell death. In the present study, mouse C2C12 muscle cells were starved in Earle’s Balanced Salt Solution or treated with TNF-α and cycloheximide to induce autophagy and apoptosis, respectively. The majority of starved C2C12 cells underwent autophagy, as shown by LC3 processing, formation of autophagic vesicles and bulk degradation of long-lived proteins. However, some cells showed features of apoptosis including caspase-3 cleavage, chromatin condensation, DNA fragmentation and annexin V labeling. Caspase-3 cleavage was also induced in culture medium without serum, suggesting that serum withdrawal rather than amino acid deprivation triggered apoptosis. Starvation eliminated multiple pro-apoptotic proteins, but upregulated caspase-8, and rendered starved C2C12 cells much more susceptible to TNF-α/cycloheximide-induced apoptosis than non-starved cells. Our data suggest that amino acid deprivation of C2C12 cells induces a complex form of cell death with hallmarks of both apoptosis and autophagy.
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References
P Anglade S Vyas F Javoy-Agid MT Herrero PP Michel J Marquez A Mouatt-Prigent M Ruberg EC Hirsch Y Agid (1997) ArticleTitleApoptosis and autophagy in nigral neurons of patients with Parkinson’s disease Histol. Histopathol 12 25–31 Occurrence Handle9046040
W Bursch H Kienzl A Ellinger L Török R Walker M Sikorska S Pandey R Schulte-Hermann (1996) ArticleTitleActive cell death induced by antiestrogens tamoxifen and ICI 164384 in human mammary carcinoma cells (MCF-7) in culture: the role of autophagy Carcinogenesis 17 1595–1607 Occurrence Handle8761415
PGH Clarke (1990) ArticleTitleDevelopmental cell death: morphological diversity and multiple mechanisms Anat. Embryol 181 195–213 Occurrence Handle10.1007/BF00174615 Occurrence Handle2186664
G Fuertes JJ Martin Llano ParticleDe A Villarroya AJ Rivett E Knecht (2003) ArticleTitleChanges in the proteolytic activities of proteasomes and lysosomes in human fibroblasts produced by serum withdrawal, amino acid deprivation and confluent conditions Biochem. J 375 75–86 Occurrence Handle10.1042/BJ20030282 Occurrence Handle12841850
S Furuta E Hidaka A Ogata S Yokota T Kamata (2004) ArticleTitleRas is involved in the negative control of autophagy through the class I PI3-kinase Oncogene 23 3898–3904 Occurrence Handle10.1038/sj.onc.1207539 Occurrence Handle15064741
D Gozuacik A Kimchi (2004) ArticleTitleAutophagy as a cell death and tumor suppressor mechanism Oncogene 23 2891–2906 Occurrence Handle10.1038/sj.onc.1207521 Occurrence Handle15077152
CA Guimaraes R Linden (2004) ArticleTitleProgrammed cell death: apoptosis and alternative deathstyles Eur. J. Biochem 271 1638–1650 Occurrence Handle10.1111/j.1432-1033.2004.04084.x Occurrence Handle15096203
JJ Lemasters A-L Nieminen T Qian LC Trost SP Elmore Y Nishimura RA Crowe WE Cascio CA Bradham DA Brenner B Herman (1998) ArticleTitleThe mitochondrial permeability transition in cell death: a common mechanism in necrosis, apoptosis and autophagy Biochim. Biophys. Acta 1366 177–196 Occurrence Handle9714796
RA Lockshin Z Zakeri (2004) ArticleTitleApoptosis, autophagy, and more Int. J. Biochem. Cell. Biol 36 2405–2419 Occurrence Handle10.1016/j.biocel.2004.04.011 Occurrence Handle15325581
N Mizushima (2004) ArticleTitleMethods for monitoring autophagy Int. J. Biochem. Cell. Biol 36 2491–2502 Occurrence Handle10.1016/j.biocel.2004.02.005 Occurrence Handle15325587
DB Munafo MI Colombo (2001) ArticleTitleA novel assay to study autophagy: regulation of autophagosome vacuole size by amino acid deprivation J. Cell. Sci 114 3619–3629 Occurrence Handle11707514
C Scaffidi S Fulda A Srinivasan C Friesen F Li KJ Tomaselli KM Debatin PH Krammer ME Peter (1998) ArticleTitleTwo CD95 (APO-1/Fas) signaling pathways EMBO J 17 1675–1687 Occurrence Handle10.1093/emboj/17.6.1675 Occurrence Handle9501089
A Tassa MP Roux D Attaix DM Bechet (2003) ArticleTitleClass III phosphoinositide 3-kinase-Beclin1 complex mediates the amino acid dependent regulation of autophagy in C2C12 myotubes Biochem. J 376 577–586 Occurrence Handle10.1042/BJ20030826 Occurrence Handle12967324
N Thorball H Moe H Winther-Nielsen (1985) ArticleTitleProgressive involution and physiological death of smooth muscle cells of rat incisal arterioles Blood Vessels 22 57–171 Occurrence Handle3978237
L Yu A Alva H Su P Dutt E Freundt S Welsh EH Baehrecke MJ Lenardo (2004) ArticleTitleRegulation of an ATG7-beclin 1 program of autophagic cell death by caspase-8 Science 304 1500–1502
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Martinet, W., De Meyer, G.R., Herman, A.G. et al. Amino Acid Deprivation Induces Both Apoptosis and Autophagy in Murine C2C12 Muscle Cells. Biotechnol Lett 27, 1157–1163 (2005). https://doi.org/10.1007/s10529-005-0007-y
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DOI: https://doi.org/10.1007/s10529-005-0007-y