Functional characteristic of PC12 cells with reduced microsomal glutathione transferase 1

• Authors: Monika Sobczak , Tomasz Boczek , Bozena Ferenc , Joanna Taha , Anna Kozaczuk , Magdalena Wiktorska , Izabela Sacewicz-Hofman , Jolanta Niewiarowska , Ludmila Zylinska
• Languages of publication: EN

Abstracts

EN

Microsomal glutathione transferase 1 (MGST1) possesses glutathione transferase and peroxidase activities and is active in biotransformation of xenobiotics and in defense against oxidative stress. To assess MGST1 role in the development and functioning of PC12 cells, we constructed a cell line with reduced MGST1 (PC12_M). Real-time PCR and immunoblot assays showed MGST1 expression lowered to 60 % and immunocytochemical analyses demonstrated an altered concentration and distribution of the enzyme. PC12_M cells revealed a larger tendency to grow in clusters, weaker adhesion, irregular shape of bodies, short neurite outgrowth and higher percentage of necrotic cells (34 %). The total GSTs activity determined with non-specific substrate CDNB (1-chloro-2,4-dinitrobenzene) decreased by 15-20 %, whereas that with DCNB (2,4-dichloro-1-nitrobenzene), a substrate more specific for cytosolic GSTs, was similar to the one in control cells. This suggests that reduction of MGST1 cannot be compensated by other glutathione transferases. In PC12_M cells the total glutathione content was higher by 15-20 %, whereas the GSSG/GSH ratio was lower than in control cells. Moreover, the laminin-dependent migration rate was much faster in control cells than in PC12_M, suggesting some alterations in the metastatic potential of the line with suppressed MGST1. The amount of MAP kinases (p38, JNK, ERK1/2) was elevated in PC12_M cells but their phosphorylation level declined. Microarray analysis showed changed expression of several genes, which may be linked with differentiation and necrosis of PC12_M cells. Our data suggest that MGST1 could be an important regulator of PC12 cells development and might have significant effects on cell growth and proliferation, probably through altered expression of genes with different biological function.

Keywords

EN

necrosis; pc12 cells; signaling pathways; microsomal glutathione transferase 1; pseudoneuronal phenotype; metastatic potential

• Publisher: Polish Biochemical Society
• Journal: Acta Biochimica Polonica
• Year: 2010
• Volume: 57
• Issue: 4
• Pages: 589-596

Dates

published

2010

received

2010-08-25

revised

2010-11-22

(unknown)

2010-12-15

accepted

2010-12-15

Contributors

author

Monika Sobczak

• Department of Molecular Neurochemistry, Medical University, Łódź, Poland

author

Tomasz Boczek

• Department of Molecular Neurochemistry, Medical University, Łódź, Poland

author

Bozena Ferenc

• Department of Molecular Neurochemistry, Medical University, Łódź, Poland

author

Joanna Taha

• Department of Molecular Neurochemistry, Medical University, Łódź, Poland

author

Anna Kozaczuk

• Department of Molecular Neurochemistry, Medical University, Łódź, Poland

author

Magdalena Wiktorska

• Department of Molecular and Medical Biophysics, Medical University, Łódź, Poland

author

Izabela Sacewicz-Hofman

• Department of Molecular and Medical Biophysics, Medical University, Łódź, Poland

author

Jolanta Niewiarowska

• Department of Molecular and Medical Biophysics, Medical University, Łódź, Poland

author

Ludmila Zylinska

• Department of Molecular Neurochemistry, Medical University, Łódź, Poland

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