title: Downregulation of microsomal glutathione-S-transferase 1 modulates protective mechanisms in differentiated PC12 cells.
abstract: Microsomal glutathione-S-transferase 1 (Mgst1) plays a specific role in protection of cells against oxidative stress. In this study, we assayed the effect of Mgst1 downregulation on cells behavior using differentiated PC12 line, a widely accepted neuronal model system. We have developed stable transfected cells with downregulated Mgst1 (PC12_M), which were differentiated with 1 mM dibutyryl-cAMP (db-cAMP). Mgst1 reduction induced necrosis, decreased ATP amount, and increased thiobarbituric acid reacting substances (TBARS) content. However, in PC12_M cell population, we detected more intensive neuritogenesis than that in mock-transfected cells. Interestingly, total glutathione as well as GSH level were significantly higher than those in control PC12 line. Real-time PCR and Western blot analyses showed elevated expression of enzymes involved in glutathione metabolism-a rate-limiting γ-glutamylcysteine ligase and glutathione reductase. The present study shows for the first time that under stress conditions induced by Mgst1 downregulation, a rescue pathway can be activated and thereby enables differentiated PC12 cells to survive. Since Mgst1expression was reported to decline with age, our results could represent a putative adaptive process during aging. It could also be an early mechanism protecting neuronal cells against some neurodegenerative insults.
authors.affiliation: Department of Molecular Neurochemistry, Medical University, 6/8 Mazowiecka Str., 92-215, Lodz, Poland.
authors.forename: Monika
authors.initials: M
authors.lastname: Sobczak
authors.forename: Tomasz
authors.initials: T
authors.lastname: Boczek
authors.forename: Antoni
authors.initials: A
authors.lastname: Kowalski
authors.forename: Magdalena
authors.initials: M
authors.lastname: Wiktorska
authors.forename: Jolanta
authors.initials: J
authors.lastname: Niewiarowska
authors.forename: Ludmila
authors.initials: L
authors.lastname: Zylinska
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D001483:Base Sequence; D002454:Cell Differentiation; D017362:Cyclic AMP Response Element-Binding Protein / Q000378:metabolism; D017931:DNA Primers; D015536:Down-Regulation; D005434:Flow Cytometry; D005982:Glutathione Transferase / Q000378:metabolism*; D008861:Microsomes / Q000201:enzymology*; D016716:PC12 Cells; D051381:Rats; D060888:Real-Time Polymerase Chain Reaction; D017392:Thiobarbituric Acid Reactive Substances / Q000378:metabolism
chemical_list: D017362:Cyclic AMP Response Element-Binding Protein; D017931:DNA Primers; D017392:Thiobarbituric Acid Reactive Substances; D000255:Adenosine Triphosphate; C122107:microsomal glutathione S-transferase-I; D005982:Glutathione Transferase