tailieunhanh - Báo cáo khoa học: Transfection with 4-hydroxynonenal-metabolizing glutathione S-transferase isozymes leads to phenotypic transformation and immortalization of adherent cells

4-Hydroxy-2-trans-nonenal (4-HNE), one of the major end products of lipid peroxidation, has been shown to induce apoptosis in a variety of cell lines. It appears to modulate signalingprocesses inmore thanonewaybecause it has been suggested to have a role in signaling for differentiation and proliferation. We showfor the first time that incorporation of 4-HNE-metabolizing glutathioneS-transferase (GST) isozyme, hGSTA4-4, into adherent cell lines HLE B-3 and CCL-75, by either cDNA transfection or microinjection of active enzyme, leads to their transformation. . | Eur. J. Biochem. 271 1690-1701 2004 FEBS 2004 doi Transfection with 4-hydroxynonenal-metabolizing glutathione 5-transferase isozymes leads to phenotypic transformation and immortalization of adherent cells Rajendra Sharma1 David Brown 1 Sanjav Awasthi2 Yusona Yana1. Abha Sharma1. Brad Patrick1 Manjit K. Saini1 Sharda P. Singh3 Piotr Zimniak3 Shivendra V. Singh4 and Yogesh C. Awasthi1 1 Department of Human Biological Chemistry and Genetics University of Texas Medical Branch Galveston TX USA department of Chemistry and Biochemistry University of Texas at Arlington TX USA 3Department of Pharmacology and Toxicology and Department of Biochemistry and Molecular Biology University of Arkansas for Medical Sciences and Central Arkansas Veterans Healthcare System Little Rock AR USA department of Pharmacology University of Pittsburgh Cancer Center PA USA 4-Hydroxy-2-írans-nonenal 4-HNE one of the major end products of lipid peroxidation has been shown to induce apoptosis in a variety of cell lines. It appears to modulate signaling processes in more than one way because it has been suggested to have a role in signaling for differentiation and proliferation. We show lor the hrtt iime that incorporaiion of 4-HNE-metabolizing glutathione S-transferase GST isozyme hGSTA4-4 into adherent cell lines HLE B-3 and CCL-75 by either cDNA transfection or microinjection of active enzyme leads to their transformation. The dramatic phenotypic changes due to the incorporation of hGSTA4-4 include rounding of cells and anchorage-independent rapid proliferation of immortalized rounded and smaller cells. Incorporation of the inactive mutant of hGSTA4-4 Y212F in cells by either microinjection or transfection does not cause transformation suggesting that the activity of hGSTA4-4 toward 4-HNE is required for transformation. This is further confirmed by the fact that mouse and Drosophila GST isozymes mGSTA4-4 and DmGSTD1-1 which have high activity toward 4-HNE and .