Gamma glutamyltransferase (GGT) is a plasma membrane bound enzyme that initiates the degradation of glutathione. The presence of several promoters in the rat GGT gene indicates strict control and regulation of its expression. The aim of this study was to investigate whether the GGT gene was regulated differently after butyrate-induced differentiation and oxidative stress exposure of rat colon carcinoma cells and whether the regulation was related to the glutathione level. The activity of GGT was upregulated in a time-and-dose dependent manner after both butyrate and menadione incubations. The presence of antioxidants blocked the menadione but not the butyrate mediated induction of the enzyme. The level of intracellular glutathione was reduced after menadione, but not after butyrate incubations. Depletion of glutathione alone did not alter GGT activity. Reactive oxygen species (ROS) were not produced after incubations with butyrate, while menadione incubations produced ROS. The multiple GGT mRNA transcripts (mRNA I–V) that originate from the five distinct promoters were all present in the cell line. Incubations with butyrate enhanced mRNA II and IV transcripts whereas a reduction in mRNA IV-1 was noted during menadione incubations. The level of total GGT mRNA (I–V) was not altered when related to the amount of total β-actin mRNA. We conclude that GGT activity can be upregulated by at least two distinct mechanisms during differentiation and oxidative stress. Apparently, the regulation of the enzyme is not directly linked to the intracellular level of glutathione.
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Hanigan MH: Expression of gamma-glutamyl transpeptidase provides tumor cells with a selective growth advantage at physiologic concentrations of cyst(e)ine. Carcinogenesis 16: 181–185, 1995
Hochwald SN, Harrison LE, Rose DM, Anderson M, Burt ME: Gammaglutamyl transpeptidase mediation of tumor glutathione utilization in vivo. J Natl Cancer Inst 88: 193–197, 1996
Karp DR, Shimoko K, Lipsky PE: Expression of γ-glutamyl transpeptidase protects ramos B cells from oxidation-induced cell death. J Biol Chem 276: 3798–3804, 2001
Cameron RG, Armstrong A, Gunsekava A, Varghese G, Speisky H: Utilization of circulating glutathione by nodular and cancerous intact rat liver. Carcinogenesis 12: 2369–2372, 1991
Varma SD, Devamanoharan PS, Ali AH: Prevention of intracellular oxidative stress to lens by pyruvate and its ester. Free Rad Res 28: 131–135, 1998
Morgan WA, Kaler B, Bach PH: The role of reactive oxygen species in adriamycin and menadione-induced glomerular toxicity. Toxicol Lett 94: 209–215, 1998
Liu RM, Shi MM, Giulivi C, Forman HJ: Quinones increase gammaglutamyl transpeptidase expression by multiple mechanisms in rat lung epithelial cells. Am J Physiol 274: L330–L336, 1998
Borud O, Mortensen B, Mikkelsen IM, Leroy P, Wellman M, Huseby NE: Regulation of gamma glutamyltransferase in cisplatin-resistant and-sensitive colon carcinoma cells after acute cisplatin and oxidative stress exposures. Int J Cancer 88: 464–468, 2000
Schroy PC, Rustgi AK, Ikonomu E, Liu XP, Polito J, Andry C, O'Keane JC: Growth and intestinal differentiation are independently regulated in HT29 colon cancer cells. J Cell Physiol 161: 111–123, 1994
Archer S, Meng S, Wu J, Johnson J, Tang R, Hodin R: Butyrate inhibits colon carcinoma cell growth through two distinct pathways. Surgery 124: 248–253, 1998
Wasserman L, Nordenberg J, Beery E, Deutsch AA, Novogrodsky A: Differential effects of sodium butyrate and dimethylsulfoxide on gamma-glutamyl transpeptidase and alkaline phosphatase activities in MCF-7 breast cancer cells. Exp Cell Biol 55: 188–193, 1987
Csordas A: On the biological role of histone acetylation. Biochem J 265: 23–38, 1990
Almouzni G, Khochbin S, Dimitrov S, Wolffe AP: Histone acetylation influences both gene expression and development of Xenopus laevis. Dev Biol 165: 654–669, 1994
Pawlak A, Lahuna O, Bulle F, Suzuki A, Ferry N, Siegrist S, Chikhi, N, Chobert MN, Guellaen G, Laperche Y: Gamma-glutamyl transpeptidase: A single copy gene in the rat and multigene family in the human genome. J Biol Chem 263: 9913–9916, 1998
Chikhi N, Holic N, Guellaen G, Laperche Y: Gamma-glutamyl transpeptidase gene organization and expression: A comparative analysis in rat, mouse, pig and human species. Comp Biochem Physiol B Biochem Mol Biol 122: 367–380, 1999
Liu RM, Hu H, Robison TW, Forman HJ: Increased gamma-glutamylcysteine synthetase and gamma-glutamyl transpeptidase activities enhance resistance of rat lung epithelial L2 cells to quinone toxicity. Am J Respir Cell Mol Biol 14: 192–197, 1996
Holic N, Suzuki T, Corlu A, Couchie D, Chobert MN, Guguen-Guillouzo C, Laperche Y: Differential expression of the rat gamma-glutamyl transpeptidase gene promoters along with differentiation of hepatoblasts into biliary or hepatocytic lineage. Am J Pathol 157: 537–548, 2000
Hudson EA, Munks RJ, Manson MM: Characterization of transcriptional regulation of gamma-glutamyl transpeptidase in rat liver involving both positive and negative regulatory elements. Mol Carcinogen 20: 376–388, 1997
Gallagher BC, Rudolph DB, Hinton BT, Hanigan MH: Differential induction of gamma-glutamyl transpeptidase in primary cultures of rat and mouse hepatocytes parallels induction during hepatocarcinogenesis. Carcinogenesis 19: 1251–1255, 1998
Marquet RL, Westbroek DL, Jeekel J: Interferon treatment of a transplantable rat colon adenocarcinoma: Importance of tumor site. Int J Cancer 33: 689–692, 1984
Grøstad M, Huseby NE: Clearance of different multiple forms of human gamma-glutamyltransferase. Clin Chem 36: 1654–1656, 1990
Brouillet A, Holic N, Chobert MN, Laperche Y: The gamma-glutamyl transpeptidase gene is transcribed from a different promoter in rat hepatocytes and biliary cells. Am J Pathol 152: 1039–1048, 1998
Siegling A, Lehmann M, Platzer C, Emmrich F, Volk HD: A novel multispecific competitor fragment for quantitative PCR analysis of cytokine gene expression in rats. Anonymous. Anonymous. J Immunol Meth 177: 23–28, 1994
Barouki R, Finidori J, Chobert MN, Aggerback M, Laperche Y, Hanoune J: Biosynthesis and processing of gamma-glutamyl transpeptidase in hepatoma tissue culture cells. J Biol Chem 259: 7970–7974, 1984
Macho A, Hirsch T, Marzo I, Marchetti P, Dallaporta B, Susin SA, Zamzami N, Kroemer G: Glutathione depletion is an early and calcium elevation is a late event of thymocyte apoptosis. J Immunol 158: 4612–4619, 1997
Ghibelli L, Fanelli C, Rotilio G, Lafavia E, Coppola S, Colussi C, Civitareale P, Ciriolo MR: Rescue of cells from apoptosis by inhibition of active GSH extrusion. FASEB J 12: 479–486, 1998
Chobert MN, Lahuna O, Lebargy F, Kurauchi O, Darbouy M, Bernaudin JF, Guellaen G, Barouki R, Laperche Y: Tissue-specific expression of two gamma-glutamyl transpeptidase mRNAs with alternative 5′ ends encoded by a single copy gene in the rat. J Biol Chem 265: 2352–2357, 1990
Brouillet A, Darbouy M, Okamoto T, Chobert MN, Lahuna O, Garlatti M, Goodspeed D, Laperche Y: Functional characterization of the rat gamma-glutamyl transpeptidase promoter that is expressed and regulated in the liver and hepatoma cells. J Biol Chem 269: 14878–14884, 1996
Oster T, Visvikis A, Thioudellet C, Fournel-Gigleux S, Wellman M, Siest G: Establishment of a V79 transfected cell line highly producing recombinant human gamma-glutamyltransferase. Toxicology 82: 151–167, 1993
Coloma J, Carcia-Jimeno A: Tissue-specific methylation in the 5′ flanking region of the gamma-glutamyl transpeptidase gene. Biochem Biophys Res Com 177: 229–234, 1991
Markey CM, Rudolph DB, Labus JC, Hinton BT: Oxidative stress differentially regulates the expression of gamma-glutamyl transpeptidase mRNAs in the initial segment of the rat epididymis. J Androl 19: 92–99, 1998
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Mikkelsen, I.M., Mortensen, B., Laperche, Y. et al. The expression of γ-glutamyltransferase in rat colon carcinoma cells is distinctly regulated during differentiation and oxidative stress. Mol Cell Biochem 232, 87–95 (2002). https://doi.org/10.1023/A:1014809607758
- sodium butyrate
- oxidative stress
- GGT mRNA isoforms
- rat colon carcinoma cells