Nucleosides block AICAR-stimulated activation of AMPK in skeletal muscle and cancer cells

AMP-activated kinase (AMPK) is a major regulator of energy metabolism and a promising target for development of new treatments for type 2 diabetes and cancer. 5-Aminoimidazole-4-carboxamide-1-β-d-ribofuranoside (AICAR), an adenosine analog, is a standard positive control for AMPK activation in cell-...

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Bibliographic Details
Published in:American Journal of Physiology: Cell Physiology 2018-12, Vol.315 (6), p.C803-C817
Main Authors: Dolinar, Klemen, Jan, Vid, Pavlin, Mojca, Chibalin, Alexander V, Pirkmajer, Sergej
Format: Article
Language:English
Subjects:
Adenosine
AMP
Ascorbic acid
Cancer
Cell activation
Cell culture
Cell proliferation
Cells
Culture media
Deoxyadenosine
Diabetes mellitus
Diabetes mellitus (non-insulin dependent)
Energy metabolism
Kinases
Lipoic acid
Medicin och hälsovetenskap
Methotrexate
Muscular system
Nucleosides
Skeletal muscle
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Summary:AMP-activated kinase (AMPK) is a major regulator of energy metabolism and a promising target for development of new treatments for type 2 diabetes and cancer. 5-Aminoimidazole-4-carboxamide-1-β-d-ribofuranoside (AICAR), an adenosine analog, is a standard positive control for AMPK activation in cell-based assays. Some broadly used cell culture media, such as minimal essential medium α (MEMα), contain high concentrations of adenosine and other nucleosides. We determined whether such media alter AICAR action in skeletal muscle and cancer cells. In nucleoside-free media, AICAR stimulated AMPK activation, increased glucose uptake, and suppressed cell proliferation. Conversely, these effects were blunted or completely blocked in MEMα that contains nucleosides. Addition of adenosine or 2'-deoxyadenosine to nucleoside-free media also suppressed AICAR action. MEMα with nucleosides blocked AICAR-stimulated AMPK activation even in the presence of methotrexate, which normally markedly enhances AICAR action by reducing its intracellular clearance. Other common media components, such as vitamin B-12, vitamin C, and α-lipoic acid, had a minor modulatory effect on AICAR action. Our findings show that nucleoside-containing media, commonly used in AMPK research, block action of the most widely used pharmacological AMPK activator AICAR. Results of cell-based assays in which AICAR is used for AMPK activation therefore critically depend on media formulation. Furthermore, our findings highlight a role for extracellular nucleosides and nucleoside transporters in regulation of AMPK activation.
ISSN:0363-6143
1522-1563
1522-1563
DOI:10.1152/ajpcell.00311.2017