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- Publisher Website: 10.1016/j.lfs.2009.01.014
- Scopus: eid_2-s2.0-61849121847
- PMID: 19302811
- WOS: WOS:000264593500012
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Article: Apoptotic signaling induced by H2O2-mediated oxidative stress in differentiated C2C12 myotubes
Title | Apoptotic signaling induced by H2O2-mediated oxidative stress in differentiated C2C12 myotubes |
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Authors | |
Keywords | Skeletal muscle Apoptotic signaling Oxidative stress Programmed cell death |
Issue Date | 2009 |
Citation | Life Sciences, 2009, v. 84, n. 13-14, p. 468-481 How to Cite? |
Abstract | Aims: Apoptotic signaling proteins were evaluated in postmitotic skeletal myotubes to test the hypothesis that oxidative stress induced by H 2 O 2 activates both caspase-dependent and caspase-independent apoptotic proteins in differentiated C2C12 myotubes. We hypothesized that oxidative stress would decrease anti-apoptotic protein levels in C2C12 myotubes. Main methods: Apoptotic regulatory factors and apoptosis-associated proteins including Bcl-2, Bax, Apaf-1, XIAP, ARC, cleaved PARP, p53, p21 Cip1/Waf1 , c-Myc, HSP70, CuZnSOD, and MnSOD protein content were measured by immunoblots. Key findings: H 2 O 2 induced apoptosis in myotubes as shown by DNA laddering and an elevation of apoptotic DNA fragmentation. Cell death ELISA showed increase in the extent of apoptotic DNA fragmentation following treatment with H 2 O 2 . Treatment with 4 mM of H 2 O 2 for 24 or 96 h caused increase in Bax (56%, 227%), cytochrome c (282%, 701%), Smac/DIABLO (155%, 260%), caspase-3 protease activity (51%, 141%), and nuclear and cytosolic p53 (719%, 1581%) levels in the myotubes. As an estimate of the mitochondrial AIF release to the cytosol, AIF protein content measured in the mitochondria-free cytosolic fraction was elevated by 65% after 96 h treatment with 4 mM of H 2 O 2 . AIF measured in the nuclear protein fraction increased by 74% and 352% following treatment with 4 mM of H 2 O 2 for 24 and 96 h, respectively. Bcl-2 declined in myotubes by 61% and 69% after 24 or 96 h of treatment in 4 mM H 2 O 2 , respectively. Significance: These findings indicate that both caspase-dependent and caspase-independent mechanisms are involved in coordinating the activation of apoptosis induced by H 2 O 2 in differentiated myotubes. © 2009 Elsevier Inc. All rights reserved. |
Persistent Identifier | http://hdl.handle.net/10722/244095 |
ISSN | 2023 Impact Factor: 5.2 2023 SCImago Journal Rankings: 1.257 |
ISI Accession Number ID |
DC Field | Value | Language |
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dc.contributor.author | Siu, Parco M. | - |
dc.contributor.author | Wang, Yan | - |
dc.contributor.author | Alway, Stephen E. | - |
dc.date.accessioned | 2017-08-31T08:56:02Z | - |
dc.date.available | 2017-08-31T08:56:02Z | - |
dc.date.issued | 2009 | - |
dc.identifier.citation | Life Sciences, 2009, v. 84, n. 13-14, p. 468-481 | - |
dc.identifier.issn | 0024-3205 | - |
dc.identifier.uri | http://hdl.handle.net/10722/244095 | - |
dc.description.abstract | Aims: Apoptotic signaling proteins were evaluated in postmitotic skeletal myotubes to test the hypothesis that oxidative stress induced by H 2 O 2 activates both caspase-dependent and caspase-independent apoptotic proteins in differentiated C2C12 myotubes. We hypothesized that oxidative stress would decrease anti-apoptotic protein levels in C2C12 myotubes. Main methods: Apoptotic regulatory factors and apoptosis-associated proteins including Bcl-2, Bax, Apaf-1, XIAP, ARC, cleaved PARP, p53, p21 Cip1/Waf1 , c-Myc, HSP70, CuZnSOD, and MnSOD protein content were measured by immunoblots. Key findings: H 2 O 2 induced apoptosis in myotubes as shown by DNA laddering and an elevation of apoptotic DNA fragmentation. Cell death ELISA showed increase in the extent of apoptotic DNA fragmentation following treatment with H 2 O 2 . Treatment with 4 mM of H 2 O 2 for 24 or 96 h caused increase in Bax (56%, 227%), cytochrome c (282%, 701%), Smac/DIABLO (155%, 260%), caspase-3 protease activity (51%, 141%), and nuclear and cytosolic p53 (719%, 1581%) levels in the myotubes. As an estimate of the mitochondrial AIF release to the cytosol, AIF protein content measured in the mitochondria-free cytosolic fraction was elevated by 65% after 96 h treatment with 4 mM of H 2 O 2 . AIF measured in the nuclear protein fraction increased by 74% and 352% following treatment with 4 mM of H 2 O 2 for 24 and 96 h, respectively. Bcl-2 declined in myotubes by 61% and 69% after 24 or 96 h of treatment in 4 mM H 2 O 2 , respectively. Significance: These findings indicate that both caspase-dependent and caspase-independent mechanisms are involved in coordinating the activation of apoptosis induced by H 2 O 2 in differentiated myotubes. © 2009 Elsevier Inc. All rights reserved. | - |
dc.language | eng | - |
dc.relation.ispartof | Life Sciences | - |
dc.subject | Skeletal muscle | - |
dc.subject | Apoptotic signaling | - |
dc.subject | Oxidative stress | - |
dc.subject | Programmed cell death | - |
dc.title | Apoptotic signaling induced by H2O2-mediated oxidative stress in differentiated C2C12 myotubes | - |
dc.type | Article | - |
dc.description.nature | link_to_subscribed_fulltext | - |
dc.identifier.doi | 10.1016/j.lfs.2009.01.014 | - |
dc.identifier.pmid | 19302811 | - |
dc.identifier.scopus | eid_2-s2.0-61849121847 | - |
dc.identifier.volume | 84 | - |
dc.identifier.issue | 13-14 | - |
dc.identifier.spage | 468 | - |
dc.identifier.epage | 481 | - |
dc.identifier.isi | WOS:000264593500012 | - |
dc.identifier.issnl | 0024-3205 | - |