Evidence for Ectopic Aerobic ATP Production on C6 Glioma Cell Plasma Membrane

Ravera, Silvia; Aluigi, Maria Grazia; Calzia, Daniela; Ramoino, Paola; Morelli, Alessandro; Panfoli, Isabella

doi:10.1007/s10571-010-9624-7

Cited by 33 publications

(25 citation statements)

References 44 publications

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“…Notably, NADH is impermeable to mitochondria. For this reason, it is useful to discriminate the extramitochondrial oxphos—for example, in rod outer segment disks (26), myelin sheath (25), and C6 glioma plasma membrane (27)—from the mitochondrial one.…”

Section: Resultsmentioning

confidence: 99%

“…ATP synthesis was assayed on preterm or term human UC MSC exosomes by the luciferin/luciferase chemiluminescent method (Roche Applied Sciences, Indianapolis, IN, USA), as reported elsewhere (26, 27). Briefly, samples were first incubated for 10 min at 37°C in 100 mM Tris‐HCl (pH 7.4), 100 mM KCl, 1 mM EGTA, 2.5 mM EDTA, 5 mM MgCl 2 , 0.2 mM di(adenosine‐5') penta‐phosphate, 0.6 mM ouabain, ampicillin (25 μg/ml), and 5 mM KH 2 PO 4 .…”

Section: Methodsmentioning

confidence: 99%

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Exosomes from human mesenchymal stem cells conduct aerobic metabolism in term and preterm newborn infants

et al. 2015

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Exosomes are secreted nanovesicles that are able to transfer RNA and proteins to target cells. The emerging role of mesenchymal stem cell (MSC) exosomes as promoters of aerobic ATP synthesis restoration in damaged cells, prompted us to assess whether they contain an extramitochondrial aerobic respiration capacity. Exosomes were isolated from culture medium of human MSCs from umbilical cord of ≥37-wk-old newborns or between 28- to 30-wk-old newborns (i.e.,term or preterm infants). Characterization of samples was conducted by cytofluorometry. Oxidative phosphorylation capacity was assessed by Western blot analysis, oximetry, and luminometric and fluorometric analyses. MSC exosomes express functional respiratory complexes I, IV, and V, consuming oxygen. ATP synthesis was only detectable in exosomes from term newborns, suggestive of a specific mechanism that is not completed at an early gestational age. Activities are outward facing and comparable to those detected in mitochondria isolated from term MSCs. MSC exosomes display an unsuspected aerobic respiratory ability independent of whole mitochondria. This may be relevant for their ability to rescue cell bioenergetics. The differential oxidative metabolism of pretermvs.term exosomes sheds new light on the preterm newborn's clinical vulnerability. A reduced ability to repair damaged tissue and an increased capability to cope with anoxic environment for preterm infants can be envisaged.-Panfoli, I., Ravera, S., Podestà, M., Cossu, C., Santucci, L., Bartolucci, M., Bruschi, M., Calzia, D., Sabatini, F., Bruschettini, M., Ramenghi, L. A., Romantsik, O., Marimpietri, D., Pistoia, V., Ghiggeri, G., Frassoni, F., Candiano, G. Exosomes from human mesenchymal stem cells conduct aerobic metabolism in term and preterm newborn infants.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Exosomes from human mesenchymal stem cells conduct aerobic metabolism in term and preterm newborn infants

et al. 2015

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“…The ATP-forming capacity at the surface of glioma cells was several times greater than that of normal cells [601] and evidence for ectopic aerobic ATP production in C6 glioma cell membranes has been presented recently [602].…”

Section: Gliomasmentioning

confidence: 99%

Purinergic signalling and cancer

Burnstock

Virgilio

2013

Purinergic Signalling

272

265

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Receptors for extracellular nucleotides are widely expressed by mammalian cells. They mediate a large array of responses ranging from growth stimulation to apoptosis, from chemotaxis to cell differentiation and from nociception to cytokine release, as well as neurotransmission. Pharma industry is involved in the development and clinical testing of drugs selectively targeting the different P1 nucleoside and P2 nucleotide receptor subtypes. As described in detail in the present review, P2 receptors are expressed by all tumours, in some cases to a very high level. Activation or inhibition of selected P2 receptor subtypes brings about cancer cell death or growth inhibition. The field has been largely neglected by current research in oncology, yet the evidence presented in this review, most of which is based on in vitro studies, although with a limited amount from in vivo experiments and human studies, warrants further efforts to explore the therapeutic potential of purinoceptor targeting in cancer.

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“…The reaction was monitored for two minutes, every 30 sec, in a luminometer (GloMax® 20/20n Luminometer, Promega Italia, Milano, Italy), by the luciferin/luciferase chemiluminescent method, with ATP standard solutions between 10 -8 and 10 -5 M (luciferin/luciferase ATP bioluminescence assay kit CLSII, Roche, Basel, Switzerland). Data were expressed as nmol ATP produced/min/10 6 cells [14]. The oxidative phosphorylation efficiency (P/O ratio) was calculated as the ratio between the concentration of produced ATP and the amount of consumed oxygen in the presence of respiring substrates and ADP.…”

Section: Fo-f1 Atp Synthase Activity Assaymentioning

confidence: 99%

Glibenclamide Mimics Metabolic Effects of Metformin in H9c2 Cells

Salani

Ravera

Fabbi

et al. 2017

Cell Physiol Biochem

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Background: Sulfonylureas, such as glibenclamide, are antidiabetic drugs that stimulate beta-cell insulin secretion by binding to the sulfonylureas receptors (SURs) of adenosine triphosphate-sensitive potassium channels (K ATP ). Glibenclamide may be also cardiotoxic, this effect being ascribed to interference with the protective function of cardiac K ATP channels for which glibenclamide has high affinity. Prompted by recent evidence that glibenclamide impairs energy metabolism of renal cells, we investigated whether this drug also affects the metabolism of cardiac cells. Methods: The cardiomyoblast cell line H9c2 was treated for 24 h with glibenclamide or metformin, a known inhibitor of the mitochondrial respiratory chain. Cell viability was evaluated by sulforodhamine B assay. ATP and AMP were measured according to the enzyme coupling method and oxygen consumption by using an amperometric electrode, while Fo-F1 ATP synthase activity assay was evaluated by chemiluminescent method. Protein expression was measured by western blot. Results: Glibenclamide deregulated energy balance of H9c2 cardiomyoblasts in a way similar to that of metformin. It inhibited mitochondrial complexes I, II and III with ensuing impairment of oxygen consumption and ATP synthase activity, ATP depletion and increased AMPK phosphorylation. Furthermore, glibenclamide disrupted mitochondrial subcellular organization. The perturbation of mitochondrial energy balance was associated with enhanced anaerobic glycolysis, with increased activity of phosphofructo kinase, pyruvate kinase and lactic dehydrogenase. Interestingly, some additive effects of glibenclamide and metformin were observed. Conclusions: Glibenclamide deeply alters cell metabolism in cardiac cells by impairing mitochondrial organization and function. This may further explain the risk of cardiovascular events associated with the use of this drug, alone or in combination with metformin.

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Evidence for Ectopic Aerobic ATP Production on C6 Glioma Cell Plasma Membrane

Cited by 33 publications

References 44 publications

Exosomes from human mesenchymal stem cells conduct aerobic metabolism in term and preterm newborn infants

Exosomes from human mesenchymal stem cells conduct aerobic metabolism in term and preterm newborn infants

Purinergic signalling and cancer

Glibenclamide Mimics Metabolic Effects of Metformin in H9c2 Cells

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