Document Detail


The role of cytochrome c on apoptosis induced by Anagrapha falcifera multiple nuclear polyhedrosis virus in insect Spodoptera litura cells.
MedLine Citation:
PMID:  22952575     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
There are conflicting reports on the role of cytochrome c during insect apoptosis. Our previous studies have showed that cytochrome c released from the mitochondria was an early event by western blot analysis and caspase-3 activation was closely related to cytochrome c release during apoptosis induced by baculovirus in Spodoptera litura cells (Sl-1 cell line). In the present study, alteration in mitochondrial morphology was observed by transmission electron microscopy, and cytochrome c release from mitochondria in apoptotic Sl-1 cells induced with Anagrapha falcifera multiple nuclear polyhedrosis virus (AfMNPV) has further been confirmed by immunofluoresence staining protocol, suggesting that structural disruption of mitochondria and the release of cytochrome c are important events during Lepidoptera insect cell apoptosis. We also used Sl-1 cell-free extract system and the technique of RNA interference to further investigate the role of cytochrome c in apoptotic Sl-1 cells induced by AfMNPV. Caspase-3 activity in cell-free extracts supplemented with exogenous cytochrome c was determined and showed an increase with the extension of incubation time. DsRNA-mediated silencing of cytochrome c resulted in the inhibition of apoptosis and protected the cells from AfMNPV-induced cell death. Silencing of expression of cytochrome c had a remarkable effect on pro-caspase-3 and pro-caspase-9 activation and resulted in the reduction of caspase-3 and caspase-9 activity in Sl-1 cells undergoing apoptosis. Caspase-9 inhibitor could inhibit activation of pro-caspase-3, and the inhibition of the function of Apaf-1 with FSBA blocked apoptosis, hinting that Apaf-1 could be involved in Sl-1 cell apoptosis induced by AfMNPV. Taken together, these results strongly demonstrate that cytochrome c plays an important role in apoptotic signaling pathways in Lepidopteran insect cells.
Authors:
Kaiyu Liu; Duanyang Shu; Na Song; Zhongchao Gai; Yuan Yuan; Juan Li; Min Li; Shuying Guo; Jianxin Peng; Huazhu Hong
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2012-08-28
Journal Detail:
Title:  PloS one     Volume:  7     ISSN:  1932-6203     ISO Abbreviation:  PLoS ONE     Publication Date:  2012  
Date Detail:
Created Date:  2012-09-06     Completed Date:  2013-02-19     Revised Date:  2013-07-12    
Medline Journal Info:
Nlm Unique ID:  101285081     Medline TA:  PLoS One     Country:  United States    
Other Details:
Languages:  eng     Pagination:  e40877     Citation Subset:  IM    
Affiliation:
Institute of Entomology, Central China Normal University, Wuhan, People's Republic of China.
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MeSH Terms
Descriptor/Qualifier:
Animals
Apoptosis*
Apoptotic Protease-Activating Factor 1 / metabolism
Caspase 3 / metabolism
Caspase 9 / metabolism
Cell-Free System
Cytochromes c / metabolism*
Dactinomycin / pharmacology
Flow Cytometry / methods
Gene Silencing
Insects
Microscopy, Fluorescence / methods
Mitochondria / metabolism
Nucleopolyhedrovirus / metabolism*
RNA Interference
RNA, Double-Stranded / metabolism
Chemical
Reg. No./Substance:
0/Apoptotic Protease-Activating Factor 1; 0/RNA, Double-Stranded; 50-76-0/Dactinomycin; 9007-43-6/Cytochromes c; EC 3.4.22.-/Caspase 3; EC 3.4.22.-/Caspase 9
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