Bupivacaine uncouples the mitochondrial oxidative phosphorylation, inhibits respiratory chain complexes I and III and enhances ROS production: Results of a study on cell cultures

Cela, Olga; Piccoli, Cláudia; Scrima, Rosella; Quarato, Giovanni; Marolla, Alessandra; Cinnella, Gilda; Dambrosio, Michele; Capitanio, Nazzareno

doi:10.1016/j.mito.2010.05.005

Cited by 83 publications

(64 citation statements)

References 56 publications

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“…Interestingly, pretreatment with 6-KCH significantly prevented ROS generation, suggesting that the ROS increase is directly associated with the uncoupling mechanism. A recent study on the anesthetic agent bupivacaine demonstrated that drug-induced mitochondrial uncoupling and ROS production can occur in HepG2 cells via inhibition of complex I and III and reverse electron transfer (59). Surprisingly, we also observed that SR4 inhibited complex I activity in crude mitochondria extracts of HepG2.…”

Section: Discussioncontrasting

confidence: 45%

SR4 Uncouples Mitochondrial Oxidative Phosphorylation, Modulates AMP-dependent Kinase (AMPK)-Mammalian Target of Rapamycin (mTOR) Signaling, and Inhibits Proliferation of HepG2 Hepatocarcinoma Cells

Figarola

Singhal

Tompkins

et al. 2015

Journal of Biological Chemistry

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Mitochondrial oxidative phosphorylation produces most of the energy in aerobic cells by coupling respiration to the production of ATP. Mitochondrial uncouplers, which reduce the proton gradient across the mitochondrial inner membrane, create a futile cycle of nutrient oxidation without generating ATP. Regulation of mitochondrial dysfunction and associated cellular bioenergetics has been recently identified as a promising target for anticancer therapy. Here, we show that SR4 is a novel mitochondrial uncoupler that causes dose-dependent increase in mitochondrial respiration and dissipation of mitochondrial membrane potential in HepG2 hepatocarcinoma cells. These effects were reversed by the recoupling agent 6-ketocholestanol but not cyclosporin A and were nonexistent in mitochondrial DNA-depleted HepG2 cells. In isolated mouse liver mitochondria, SR4 similarly increased oxygen consumption independent of adenine nucleotide translocase and uncoupling proteins, decreased mitochondrial membrane potential, and promoted swelling of valinomycin-treated mitochondria in potassium acetate medium. Mitochondrial uncoupling in HepG2 cells by SR4 results in the reduction of cellular ATP production, increased ROS production, activation of the energy-sensing enzyme AMPK, and inhibition of acetyl-CoA carboxylase and mammalian target of rapamycin signaling pathways, leading to cell cycle arrest and apoptosis. Global analysis of SR4-associated differential gene expression confirms these observations, including significant induction of apoptotic genes and down-regulation of cell cycle, mitochondrial, and oxidative phosphorylation pathway transcripts at 24 h post-treatment. Collectively, our studies demonstrate that the previously reported indirect activation of AMPK and in vitro anticancer properties of SR4 as well as its beneficial effects in both animal xenograft and obese mice models could be a direct consequence of its mitochondrial uncoupling activity. Hepatocellular carcinoma (HCC)2 is the most common and severe form of liver cancer, accounting for about 80 -90% of primary liver cancers and 5% of all human cancers. More than 600,000 deaths are attributed to HCC every year with 2:1 ratio for men versus women (1, 2). HCC is a primary cancer of hepatocytes that most typically occurs in the setting of known risk factors, including cirrhosis and chronic hepatitis B virus or hepatitis C virus infections (2, 3), although recently, several lines of evidence suggest that type 2 diabetes is also an independent risk factor for HCC development (4). HCC is an aggressive tumor and represents a major health problem as its incidence is increasing. Systemic chemotherapies have proven ineffective against advanced HCC, so it typically leads to death within 6 -20 months (5). Hepatocarcinogenesis is a multistep process involving inflammation, hyperplasia, and dysplasia that finally leads to malignant transformation. In recent years, mitochondria have been found to provide a novel targeting site for new anticancer drugs (known as "mitocans") that ca...

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Section: Discussioncontrasting

confidence: 45%

SR4 Uncouples Mitochondrial Oxidative Phosphorylation, Modulates AMP-dependent Kinase (AMPK)-Mammalian Target of Rapamycin (mTOR) Signaling, and Inhibits Proliferation of HepG2 Hepatocarcinoma Cells

Figarola

Singhal

Tompkins

et al. 2015

Journal of Biological Chemistry

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“…The specific activities of each complex of the respiratory chain (CI-CV) were assayed spectrophotometrically on frozen–thawed and ultrasound-treated cells as previously described [39; 40]. …”

Section: Methodsmentioning

confidence: 99%

Rewiring carbohydrate catabolism differentially affects survival of pancreatic cancer cell lines with diverse metabolic profiles

et al. 2017

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An increasing body of evidence suggests that targeting cellular metabolism represents a promising effective approach to treat pancreatic cancer, overcome chemoresistance and ameliorate patient's prognosis and survival. In this study, following whole-genome expression analysis, we selected two pancreatic cancer cell lines, PANC-1 and BXPC-3, hallmarked by distinct metabolic profiles with specific concern to carbohydrate metabolism. Functional comparative analysis showed that BXPC-3 displayed a marked deficit of the mitochondrial respiratory and oxidative phosphorylation activity and a higher production of reactive oxygen species and a reduced NAD+/NADH ratio, indicating their bioenergetic reliance on glycolysis and a different redox homeostasis as compared to PANC-1. Both cell lines were challenged to rewire their metabolism by substituting glucose with galactose as carbon source, a condition inhibiting the glycolytic flux and fostering full oxidation of the sugar carbons. The obtained data strikingly show that the mitochondrial respiration-impaired-BXPC-3 cell line was unable to sustain the metabolic adaptation required by glucose deprivation/substitution, thereby resulting in a G2\M cell cycle shift, unbalance of the redox homeostasis, apoptosis induction. Conversely, the mitochondrial respiration-competent-PANC-1 cell line did not show clear evidence of cell sufferance. Our findings provide a strong rationale to candidate metabolism as a promising target for cancer therapy. Defining the metabolic features at time of pancreatic cancer diagnosis and likely of other tumors, appears to be crucial to predict the responsiveness to therapeutic approaches or coadjuvant interventions affecting metabolism.

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“…As a result, we suppose that ClO 4 -exert its effect by perturbing the oxidoreduction chemistry of the quinone binding site causing leakage of electrons to O 2 with formation of the superoxide anion radical largely from complexes I (Dröse et al 2008). Indeed, ROS are generated by ClO 4 --dependent perturbation of the respiratory chain complexes I, but are also causally linked to their inhibition (Cela et al 2010).…”

Section: Discussionmentioning

confidence: 99%

“…Complex II activity was measured by the rate of 2,6-DCPIP reduction at 600 nm (Albayrak et al 2003;Lemarie et al 2009). Complex III activity was measured by following the initial antymicin A-sensitive rate of cytochrome c reduction at 550 nm in the presence of rotenone and decylubiquinone (Cela et al 2010). Complex IV activity was assayed by following the decrease in absorbance due to the oxidation of previously reduced cytochrome c at 550 nm according to the method described by Rustin (1994).…”

Section: Measurement Of the Mitochondrial Respiratory Chain Complexesmentioning

confidence: 99%

Perchlorate-induced oxidative stress in isolated liver mitochondria

et al. 2014

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As a new threat to environment all through the world, perchlorate (ClO4(-)) was predominantly a thyrotoxin, and its toxic manifestations in non-thyroid were also documented. To date, little is known about the effect of ClO4(-) on cell and organelle. To reveal the toxicity of ClO4(-) on living organism in-depth, mitochondria isolated from liver of Carassius auratus were incubated with different concentrations of ClO4(-). The results demonstrated that ClO4(-)-induced mitochondrial oxidative stress, and subsequently caused a gradual opening of permeability transition pore leading to mitochondrial swelling and lipid peroxidative membrane damage. ClO4(-) has a conspicuous inhibition of electron transport chain activity which largely correlated to complexes I and IV. The investigations clearly demonstrated the oxidative stress of ClO4(-) in mitochondria, may well reveal cytotoxic effects in vitro that merit further investigation.

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Bupivacaine uncouples the mitochondrial oxidative phosphorylation, inhibits respiratory chain complexes I and III and enhances ROS production: Results of a study on cell cultures

Cited by 83 publications

References 56 publications

SR4 Uncouples Mitochondrial Oxidative Phosphorylation, Modulates AMP-dependent Kinase (AMPK)-Mammalian Target of Rapamycin (mTOR) Signaling, and Inhibits Proliferation of HepG2 Hepatocarcinoma Cells

SR4 Uncouples Mitochondrial Oxidative Phosphorylation, Modulates AMP-dependent Kinase (AMPK)-Mammalian Target of Rapamycin (mTOR) Signaling, and Inhibits Proliferation of HepG2 Hepatocarcinoma Cells

Rewiring carbohydrate catabolism differentially affects survival of pancreatic cancer cell lines with diverse metabolic profiles

Perchlorate-induced oxidative stress in isolated liver mitochondria

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