Document Detail

Identification of the proteins related to cytochrome P450 induced by fenvalerate in a Trichoplusia ni cell line.
MedLine Citation:
PMID:  17484068     Owner:  NLM     Status:  MEDLINE    
In order to reveal the metabolic reaction to the presence of fenvalerate mediated by P450 in insects, we used the trypan blue exclusion technique and 3-(4,5-dimethylthiazol)-2,5-diphenyltrazolium bromide (MTT) reduction assay to assess the vitality of Trichoplusia ni (Tn) cells treated with fenvalerate, and observed dose- and time-dependent changes in total cellular P450s. In addition, two-dimensional electrophoresis (2-DE) and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) were used to identify the proteins involved in the fenvalerate reaction process. Finally, the cDNA of P450 fragments was cloned and real-time RT-PCR was performed. Our data showed that at the 0-15 mumol/L challenge concentration of fenvalerate, at which the vitality of Tn cells was not affected (p > 0.05), there was a tendency toward a dose- and time-response of total cellular P450s, which peaked at the 9 h (p < 0.05) and 12 h (p < 0.01) time points following 12.5 mumol/L stimulation with fenvalerate. The 2-DE assay detected more than 1300 protein spots in each two-dimensional gel, of which 33 spots displayed significant differences. Among the changed spots, three isoforms of P450 were identified. One of the three P450 cDNA fragments (CYP4L4) was cloned and sequenced, and its expression in treated Tn cells increased significantly (p < 0.01). It was found that fenvalerate induced the expression of P450s in insect cells. This suggests that fenvalerate could be metabolized by CYP4L4 through a hydroxylation reaction in insect cells.
Xin-Kui Fang; Dan-Feng Huang; Zhao-Xia Wang; Chun-Ling Wan; Tao Sun; Wang-Jie Xu; Chun-Yan Liu; Pei Zhou; Zhong-Dong Qiao
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2007-05-07
Journal Detail:
Title:  Cell biology and toxicology     Volume:  23     ISSN:  0742-2091     ISO Abbreviation:  Cell Biol. Toxicol.     Publication Date:  2007 Nov 
Date Detail:
Created Date:  2008-01-01     Completed Date:  2008-03-06     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  8506639     Medline TA:  Cell Biol Toxicol     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  445-57     Citation Subset:  IM    
School of Agriculture and Biology, Shanghai Jiaotong University, 800 Dongchuan Road, 200240 Shanghai, PR China.
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MeSH Terms
Cell Line
Cell Survival / drug effects
Cytochrome P-450 Enzyme System / genetics,  metabolism*
Electrophoresis, Gel, Two-Dimensional
Enzyme Induction / drug effects
Insect Proteins / analysis*,  chemistry
Moths / cytology*,  drug effects,  enzymology*
Nitriles / pharmacology*
Pyrethrins / pharmacology*
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Reg. No./Substance:
0/Insect Proteins; 0/Nitriles; 0/Pyrethrins; 0/cytochrome P-450 CYP4L4 (cabbage armyworm); 51630-58-1/fenvalerate; 9035-51-2/Cytochrome P-450 Enzyme System

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