Depletion of phospholipid hydroperoxide glutathione peroxidase up‐regulates arachidonate metabolism by 12(S)‐lipoxygenase and cyclooxygenase 1 in human epidermoid carcinoma A431 cells

Autor: Wen Chang Chang, Ching Jiunn Chen, Huei Sheng Huang
Rok vydání: 2003
Předmět:
Zdroj: The FASEB Journal. 17:1694-1696
ISSN: 1530-6860
0892-6638
DOI: 10.1096/fj.02-0847fje
Popis: Phospholipid hydroperoxide glutathione peroxidase (PHGPx), a selenium-dependent glutathione peroxidase, can interact with lipophilic substrates, including the phospholipid hydroperoxides, fatty-acid hydroperoxides, and cholesteryl ester hydroperoxides, and reduce them to hydroxide compounds. We studied the functional role of endogenous PHGPx in regulation of 12(S)-lipoxygenase and cyclooxygenase 1 activities in human epidermoid carcinoma A431 cells by using a cell system overexpressing anti-PHGPx mRNA. A retroviral expression vector designated as L1-3, wherein cDNA of PHGPx was reversely inserted into pFB-ERV in antisense orientation, was constructed. A number of stable transfectants of A431 cells with PHGPx depletion were generated from virions containing plasmid L1-3. In an intact cell assay system, the metabolism of arachidonic acid to prostaglandin E2 and 12(S)-hydroxyeicosatetraenoic acid was significantly enhanced in stable L1-3 transfectants compared with that in vector-control cells. Flow cytometric analysis revealed a significant elevated level of intracellular hydroperoxides in stable L1-3 transfectants. Treatment of stable L1-3 transfectants with 50 microM arsenite induced more significant formation of intracellular hydroperoxides than that of vector-control cells. Taken together, these results support the notion that the endogenous PHGPx plays a pivotal role in the regulation of 12(S)-lipoxygenase and cyclooxygenase 1 activities by reducing the level of intracellular lipid hydroperoxides in arachidonate metabolism in A431 cells.
Databáze: OpenAIRE
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