There is no simple method to maintain a constant ethanol concentration in long-term cell culture: Keys to a solution applied to the survey of astrocytic ethanol absorption

Eysseric, Hélène; Gonthier, B.; Soubeyran, A.; Bessard, Germain; Saxod, Raymond; Barret, L.

doi:10.1016/s0741-8329(96)00112-7

Cited by 42 publications

(43 citation statements)

References 12 publications

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“…The 0.5% ethanol lies in the toxic range but can also be achieved physiologically, particularly in chronic alcohol users. In addition, ethanol is volatile, and the amount that remains would be significantly less than what was added to cell culture medium (20). These data, therefore, suggest that ethanol can enhance complete HCV replication, at physiologically attainable concentrations.…”

Section: Ethanol Increases the Completementioning

confidence: 87%

Ethanol Enhances Hepatitis C Virus Replication through Lipid Metabolism and Elevated NADH/NAD+

Seronello

Ito

Wakita

et al. 2010

Journal of Biological Chemistry

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Ethanol has been suggested to elevate HCV titer in patients and to increase HCV RNA in replicon cells, suggesting that HCV replication is increased in the presence and absence of the complete viral replication cycle, but the mechanisms remain unclear. In this study, we use Huh7 human hepatoma cells that naturally express comparable levels of CYP2E1 as human liver to demonstrate that ethanol, at subtoxic and physiologically relevant concentrations, enhances complete HCV replication. The viral RNA genome replication is affected for both genotypes 2a and 1b. Acetaldehyde, a major product of ethanol metabolism, likewise enhances HCV replication at physiological concentrations. Ethanol consumption is a well-known risk factor for chronic liver diseases. Ethanol is also a key cofactor in the pathogenesis induced by hepatitis C virus (HCV), 2 and it decreases the efficacy of anti-HCV treatments (1, 2). Likewise, HCV infection exacerbates liver damage caused by prolonged alcohol abuse (2). It has also been reported that patients with a history of alcohol abuse are more likely to be infected with HCV than the rest of the population (1). In addition, ethanol has been suggested to exacerbate HCVinduced liver diseases in part by affecting the viral titer (2-5). Hepatitis C patients who drink alcohol typically show a pattern of hepatic injury that is more characteristic of chronic viral hepatitis than alcohol-induced injury, suggesting that alcohol enhances the pathogenic effects of HCV rather than exerting its independent effects on liver (6). Several clinical studies have correlated increased serum and intrahepatic HCV titer with the amount of alcohol consumed (2, 4, 5). Abstinence or moderation of alcohol consumption could reduce the HCV titer in some patients (2, 5). Furthermore, in vitro studies suggest that ethanol increases HCV RNA levels in Huh7 human hepatoma replicon cell lines that continuously support the HCV RNA replication without virus production (3,7,8). These studies suggest that ethanol enhances HCV replication both in the presence and absence of the complete viral replication cycle. HCV replicon systems and more recent virus-producing cell culture models have increased our understanding of HCV and provide us with tools for studying potential interactions between HCV and pathological cofactors, such as ethanol (9).Nevertheless, whether ethanol directly enhances HCV production in the context of the complete viral replication cycle has not been demonstrated. Furthermore, the mechanism by which ethanol modulates HCV RNA replication remains controversial as reactive oxygen species (ROS) and lipid peroxidation products, which can be generated during ethanol metabolism, can suppress, rather than increase, HCV RNA replication in cells, suggesting the involvement of other metabolites of ethanol (10 -14). Oxidative hepatic ethanol metabolism is a multistep process (4). Alcohol dehydrogenase, the predominant ethanol-metabolizing enzyme, is found in the cytosol and produces acetaldehyde and NADH. Ethanol-inducible cyto...

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Section: Ethanol Increases the Completementioning

confidence: 87%

Ethanol Enhances Hepatitis C Virus Replication through Lipid Metabolism and Elevated NADH/NAD+

Seronello

Ito

Wakita

et al. 2010

Journal of Biological Chemistry

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“…1A), with 5 mL of medium, until the cells were 80% confluent. Ethanol-treated cells (25 and 50 mM) were cultured in an ethanol-saturated incubator as previously described [27]. These concentrations are equivalent to the expected in the human placental tissue from moderate ethanol consumers (5e90 mg/dl; approximately 20 mM) [28] and in alcohol-depending consumers (386 mg/dl; approximately 85 mM) [29].…”

Section: Experimental Designmentioning

confidence: 99%

Prenatal ethanol exposure and placental hCG and IGF2 expression

et al. 2015

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“…For the volume of the boxes used in this study (2.016 Â 10 À3 m 3 ), the amount of ethanol required in the open dish was 1.5 mmol, according to a previous article [19]. The ethanol concentration in the media was checked after 24 h with a Perkin-Elmer Gas Chromatograph F-22, using n-propanol as an internal standard [20].…”

Section: Establishment Of Chronic Ethanol-treated Hep 3b Cell Linesmentioning

confidence: 99%

Long-term ethanol exposure causes human liver cancer cells to become resistant to mitomycin C treatment through the inactivation of bad-mediated apoptosis

et al. 2010

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The aim of this study was to test whether long-term ethanol consumption confers therapeutic resistance to human liver cancer patients infected with hepatitis B virus (HBV). Chronic ethanol-treated cells were established by consecutively culturing a human hepatocellular carcinoma cell line, Hep 3B, which contains integrated HBV sequences, for 20-40 passages with or without 10 mM ethanol (designated as E20-E40 and C20-C40, respectively). Flow cytometry analysis demonstrated that a growth promoting effect of long-term ethanol treatment was induced in the E40 cells through preferential acceleration of S-phase in these cells. Lower protein expression levels of p16, p21/Cip1, and p27/Kip1 were detected in the ethanol-treated E40 cells. We further demonstrated that long-term ethanol-treated E40 cells develop drug resistance in response to mitomycin C (MMC) treatment (>8 microM). Immunoblot analysis revealed that caspase-8-mediated mitochondrial apoptotic signals (such as Bad) were inactivated in the MMC-resistant E40 cells. Immunoprecipitation experiments demonstrated that the sequestration of phosphorylated Bad (Ser-112) through its binding with 14-3-3 was detected more profoundly in the MMC-resistant E40 cells. Next, we examined the therapeutic efficacy of MMC (10 mg MMC/kg body weight, three times per week) in severe combined immunodeficient (SCID) mice bearing E40- and C40-xenografted tumors. Significant reductions (>3-fold) in tumor growth were detected in MMC-treated C40-xenografted mice. In vivo and in vitro studies demonstrated that AKT- and extracellular signal-regulated kinase (ERK)-mediated survival factors inhibited the Bad-induced mitochondrial apoptotic signals that were involved in E40 tumor cells and that conferred resistance to MMC.

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There is no simple method to maintain a constant ethanol concentration in long-term cell culture: Keys to a solution applied to the survey of astrocytic ethanol absorption

Cited by 42 publications

References 12 publications

Ethanol Enhances Hepatitis C Virus Replication through Lipid Metabolism and Elevated NADH/NAD+

Ethanol Enhances Hepatitis C Virus Replication through Lipid Metabolism and Elevated NADH/NAD+

Prenatal ethanol exposure and placental hCG and IGF2 expression

Long-term ethanol exposure causes human liver cancer cells to become resistant to mitomycin C treatment through the inactivation of bad-mediated apoptosis

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