The phosphoenolpyruvate carboxykinase (PEPCK) inhibitor, 3-mercaptopicolinic acid (3-MPA), induces myogenic differentiation in C2C12 cells
Autor: | John M. Brameld, Paul T. Loughna, Zoe Daniel, Madelaine C. Brearley, Tim Parr |
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Rok vydání: | 2020 |
Předmět: |
Gene isoform
Science Muscle Development Article Cell Line Serine Mice chemistry.chemical_compound PCK1 PCK2 Animals Myocyte RNA Messenger Enzyme Inhibitors Picolinic Acids Promoter Regions Genetic Cells Cultured Cell Proliferation Muscle Cells Multidisciplinary biology Gluconeogenesis Cell Differentiation Molecular biology Enzyme assay Isoenzymes Metabolism Gene Expression Regulation chemistry Differentiation Phosphoserine biology.protein Medicine Phosphoenolpyruvate Carboxykinase (GTP) Phosphoenolpyruvate carboxykinase Biomarkers Phosphoenolpyruvate Carboxykinase (ATP) |
Zdroj: | Scientific Reports Scientific Reports, Vol 10, Iss 1, Pp 1-10 (2020) |
ISSN: | 2045-2322 |
Popis: | Phosphoenolpyruvate carboxykinase (PEPCK) is a gluconeogenic enzyme with a cytosolic (Pck1/PEPCK-C) and mitochondrial (Pck2/PEPCK-M) isoform. Here we investigate the effect of 3-mercaptopicolinic acid (3-MPA), a PEPCK inhibitor, on C2C12 muscle cells. We report that Pck2 mRNA is 50–5000-fold higher than Pck1 during C2C12 myogenesis, indicating Pck2 is the predominant PEPCK isoform. C2C12 cell proliferation was inhibited in a dose-dependent manner following 48 h 3-MPA treatment (0.01–1 mM). C2C12 myogenic differentiation was significantly induced following 3-MPA treatment (0.25, 0.5, 1 mM) from day 0 of differentiation, demonstrated by increased creatine kinase activity, fusion index and myotube diameter; likewise, the myosin heavy chain (MyHC)-IIB isoform (encoded by Myh4) is an indicator of hypertrophy, and both porcine MYH4-promoter activity and endogenous Myh4 mRNA were also significantly induced. High doses (0.5 and/or 1 mM) of 3-MPA reduced mRNA expression of Pck2 and genes associated with serine biosynthesis (Phosphoglycerate dehydrogenase, Phgdh; phosphoserine aminotransferase-1, Psat1) following treatment from days 0 and 4. To conclude, as Pck2/PEPCK-M is the predominant isoform in C2C12 cells, we postulate that 3-MPA promoted myogenic differentiation through the inhibition of PEPCK-M. However, we were unable to confirm that 3-MPA inhibited PEPCK-M enzyme activity as 3-MPA interfered with the PEPCK enzyme assay, particularly at 0.5 and 1 mM. |
Databáze: | OpenAIRE |
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