Inhibition of Arachidonate Metabolism in Human Epidermoid Carcinoma A431 Cells Overexpressing Phospholipid Hydroperoxide Glutathione PeroxidaseChen C.J. · Huang H.S. · Chang W.C.
Departments of aPharmacology and bMedical Technology, College of Medicine, National Cheng Kung University, Tainan, Taiwan, ROC
Phospholipid hydroperoxide glutathione peroxidase (PHGPx), a selenium-dependent glutathione peroxidase, can interact with lipophilic substrates, including phospholipid hydroperoxides, fatty acid hydroperoxides and cholesterol hydroperoxides, and can reduce them to hydroxide compounds. It also seems to be a major regulator of lipid oxygenation in human epidermoid carcinoma A431 cells. In order to study the functional role of PHGPx in the regulation of 12-lipoxygenase and cyclooxygenase, cDNA of PHGPx was inserted into pcDNA3.1/His, and a plasmid designated as S4 with the His-tag sequence inserted between PHGPx and its 3′-untranslated region was constructed. A number of stable transfectants of A431 cells that could express the tag-PHGPx were generated using plasmid S4. Using an intact cell assay system, the metabolism of arachidonic acid to prostaglandin E2 significantly decreased in stable transfectants of overexpressing PHGPx compared to that in a vector control cell line. If the intact cell assay was carried out in the presence of 13-hydroperoxyoctadecadienoic acid as a stimulator of lipid peroxidation, formation of 12-hydroxyeicosatetraenoic acid from arachidonic acid also significantly decreased in stable transfectants of overexpressing PHGPx compared to that in a vector control cell line, indicating that PHGPx could downregulate the 12-lipoxygenase activity in cells. These results support the hypothesis that PHGPx plays a pivotal role in the regulation of arachidonate metabolism in A431 cells.
Wen-Chang Chang, PhD, Department of Pharmacology
College of Medicine, National Cheng Kung University
138 Sheng-Li Road, Tainan 704, Taiwan (ROC)
Tel. +886 6 235 3535 (ext. 5496), Fax +886 6 274 9296
Received: Received: March 22, 2002
Accepted: April 3, 2002
Number of Print Pages : 7
Number of Figures : 6, Number of Tables : 0, Number of References : 25
Journal of Biomedical Science (Sponsored by the National Science Council, Taipei)
Vol. 9, No. 5, Year 2002 (Cover Date: September-October 2002)
Journal Editor: S.H.H.Chan, Kaohsiung
ISSN: 1021–7770 (print), 1423–0127 (Online)
For additional information: http://www.karger.com/journals/jbs