Use of Cultured Hepatoma Cell Lines in the Assessment of Aldehyde Metabolism

Aldehyde dehydrogenases (ALDH) represent a major pathway for the enzymatic removal of many potentially toxic aldehydes. The purpose of this study was to examine the basal levels of ALDH in five hepatoma cell lines chosen as representatives of three different species (man, rat, mouse) and their inducibility by some xenobiotics. Human HepG2, rat MH1C1, HTC, H4IIEC3 and mouse Hepa 1c1c7 cell lines were grown as monolayers. The ALDH activities were determined in cell homogenates from both unexposed control cultures and cells exposed to phenobarbital (PB), 3-methylcholanthrene (MC) and β-naphthoflavone (BNF). The ALDH activity was detected using benzaldehyde (BA) and propionaldehyde (PA) as substrates and both NAD and NADP as co-enzymes. Great variability in basal ALDH levels was found in the five cell lines: BA/NAD and BA/NADP enzyme activities were very high in the HTC cell line, intermediate in MH1C1 cells (near to normal rat hepatocytes) and very low in the remaining three cell lines. In HTC cells only, the PA/NAD activity was slightly induced by PB, but it remained unchanged under all the other experimental conditions. MH1C1 cells showed highly significant increases of all the activities with MC and BNF (up to 10-fold). The low basal activity of the H4IIEC3 cell line was significantly increased by MC and BNF, but only with BA/NADP. The Hepa 1c1c7 cell line responded only to BNF, as inducing compound, whereas the low basal enzyme levels of the human-derived HepG2 cell line were not significantly increased. These results suggest various applications of hepatoma cell cultures in in vitro toxicology.

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Evaluation of the Cytotoxicity of the First 20 MEIC Chemicals in Two Hepatoma Cell Lines with Different Xenobiotic Metabolism Capacities

Alternatives to Laboratory Animals ◽

10.1177/026119299302100111 ◽

1993 ◽

Vol 21 (1) ◽

pp. 65-72

Author(s):

Anna Maria Bassi ◽

Ornella Bosco ◽

Sabrina Brenci ◽

Daniela Adamo ◽

Susanna Penco ◽

...

Keyword(s):

Cell Line ◽

Cell Lines ◽

Hepatoma Cell ◽

Biochemical Properties ◽

Hep G2 ◽

Experimental Conditions ◽

Hepatoma Cell Lines ◽

Wide Range

The cytotoxicity of the first 20 chemicals on the MEIC list was evaluated using two hepatoma cell lines having different xenobiotic metabolic capacities, one derived from the rat (HTC), and one of human origin (Hep G2). Two endpoints were measured to evaluate cytotoxicity: colony-forming ability (CF), and cell viability of the attached monolayer (CV), evaluated as total macromolecular content. The CF test provided the most sensitive endpoint, due to the lower number of exposed cells in comparison with the CV test. Using the CF assay, the Hep G2 cell line showed higher sensitivity than the HTC cell line to some chemicals known to be metabolised in vivo. The IC50s obtained under the different experimental conditions varied as a consequence of phenotypic differences between the cell lines, and of the nature of the endpoints. The most toxic chemicals were malathion, diazepam and amitriptyline (IC50: 0.001-0.1mM). The solvents tested produced the lowest toxic effects (IC50: > 10mM). These findings suggest that hepatoma cell lines possessing various specific enzyme activities could be usefully employed in a battery of tests designed to reproduce in vitro the wide range of biochemical properties expressed by the cells in the whole organism.

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Looking for synergy or additivity between 188Re and sorafenib on hepatoma cell lines.

Journal of Clinical Oncology ◽

10.1200/jco.2012.30.4_suppl.247 ◽

2012 ◽

Vol 30 (4_suppl) ◽

pp. 247-247

Author(s):

Marc Pracht ◽

Nicolas Lepareur ◽

Julien Edeline ◽

Laurence Lenoir ◽

Valerie Ardisson ◽

...

Keyword(s):

Cell Line ◽

Hepg2 Cell ◽

Cell Lines ◽

Hepatoma Cell ◽

External Beam Radiotherapy ◽

Combined Treatment ◽

Hepatoma Cell Lines ◽

Heparg Cell Line

247 Background: In case of non resectable HCC, radioembolization and sorafenib (S) are therapeutic options respectively for intermediate and advanced stages. In some other cancers, there is an increase of efficacy when external beam radiotherapy is done concomitantly with systemic chemotherapy or targeted therapies. So we wondered if there could be a synergistic or an additive activity when S is combined with a radionuclide. Methods: Hepatoma cell lines N1S1 (murine HCC), HepG2 (human hepatoblastoma) and HepaRG (human HCC) were treated with increasing concentrations of rhenium-188 (188Re) or S. On each cell line, we have studied the cellular toxicities of S and 188Re using Tetrazolium dye test, extra-cellular medium LDH level and morphologic analysis. This was done for different dosage of S and 188Re. We measured the lethal concentration killing 25% of cells (LC25) with the results of the Tetrazolium dye test. Secondly, we looked for synergy or additivity on cellular toxicity of these two compounds according to cell lines by combined treatment. Synergy or additivity was estimated with the combination index (CI) method (synergy if CI lower than 1, additivity if CI = 1, antagonism if CI upper to 1) based on the Tetrazolium dye test’s results. Results: Monotherapy dose-dependent toxicities were observed for all three cell lines with 188Re and for the N1S1 and HepG2 cell lines only with S. Combined treatment with 188Re and S showed synergy on HepaRG and N1S1 cell lines and additivity on the HepG2 cell line. Conclusions: The additive, and even synergistic, interest of a combined treatment with 188Re and S is demonstrated in vitro (for the first time to our knowledge) on hepatoma cell lines. This results, in particular for the HepaRG cell line (human HCC), could be explained by the down-regulation of the hepatic drug transporters which are responsible for the Sorafenib efflux in case of simultaneous DNA damages due to a radionuclide exposition. This promising approach now needs to be confirmed in vivo. [Table: see text]

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Non-genotoxic hepatocarcinogenesis in vitro: the FaO hepatoma line responds to peroxisome proliferators and retains the ability to undergo apoptosis

Journal of Cell Science ◽

10.1242/jcs.104.2.307 ◽

1993 ◽

Vol 104 (2) ◽

pp. 307-315 ◽

Cited By ~ 1

Author(s):

A.C. Bayly ◽

N.J. French ◽

C. Dive ◽

R.A. Roberts

Keyword(s):

Cell Death ◽

Cell Line ◽

Cell Lines ◽

Hepatoma Cell ◽

Hepatoma Cell Line ◽

Peroxisome Proliferator ◽

Peroxisome Proliferators ◽

Hepatoma Cell Lines

A range of hepatoma cell lines (RH1, HTC, FaO, 7800C1 and MH1C1), has been studied with the aim of establishing an in vitro model to investigate the molecular mechanisms of hepatocarcinogenicity induced by the peroxisome proliferator class of non-genotoxic carcinogens. In view of speculation that peroxisome proliferators suppress hepatocyte apoptosis in vivo, we have placed particular emphasis on evaluating whether hepatoma cell lines retain the ability to undergo apoptotic cell death. Expression of the liver-specific differentiation marker albumin and the peroxisome proliferator-activated receptor (PPAR) was highest in the Reuber hepatoma cell line, FaO. This cell line also demonstrated the most marked response to the peroxisome proliferator nafenopin with a 2.2-fold induction of the microsomal enzyme cytochrome p450IVA1. This response was found to display intercellular heterogeneity by immunocytochemistry. Thus, the FaO cell line maintained characteristics of hepatocytes, both in vivo and in vitro, in terms of expression of constitutive and inducible markers. However, none of the cell lines tested mirrored the hyperplastic response of hepatocytes to nafenopin, since no increase in cell growth kinetics was observed on addition of nafenopin to the growth medium. The mode of cell death in confluent FaO cultures was characterised as apoptosis, by fluorescence microscopy and agarose gel electrophoresis of extracted DNA. Cells detaching from confluent FaO cultures exhibited chromatin condensation and DNA fragmentation patterns characteristic of cels undergoing apoptotic death.Interestingly, no apoptosis was seen in monolayer cells, suggesting that apoptosis in vitro is associated with cell shrinkage and detachment similar to that documented for the liver in vivo.(ABSTRACT TRUNCATED AT 250 WORDS)

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Differential Expression of Five Protein Kinase C Isoenzymes in FAO and HEPG2 Hepatoma Cell Lines Compared with Normal Rat Hepatocytes

Biochemical and Biophysical Research Communications ◽

10.1006/bbrc.1995.2810 ◽

1995 ◽

Vol 217 (2) ◽

pp. 546-553 ◽

Cited By ~ 33

Author(s):

L. Ducher ◽

F. Croquet ◽

S. Gil ◽

J. Davy ◽

J. Feger ◽

...

Keyword(s):

Protein Kinase C ◽

Protein Kinase ◽

Differential Expression ◽

Cell Lines ◽

Hepatoma Cell ◽

Rat Hepatocytes ◽

Hepatoma Cell Lines ◽

Kinase C ◽

Normal Rat ◽

Hepg2 Hepatoma Cell

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Different Expression of Esterase Variants in Rat Hepatic and Hepatom a-Derived Cell Lines Detected by Electrophoresis

Zeitschrift für Naturforschung C ◽

10.1515/znc-1995-9-1011 ◽

1995 ◽

Vol 50 (9-10) ◽

pp. 664-668 ◽

Cited By ~ 2

Author(s):

Adel S. Afify ◽

Yoshimitsu Yamazaki ◽

Yu-ichi Kageyama ◽

Shiro Yusa ◽

Yoshikatsu Ogawa ◽

...

Keyword(s):

Cell Lines ◽

Rat Liver ◽

Hepatoma Cell ◽

Polyacrylamide Gel Electrophoresis ◽

Liver Homogenate ◽

Normal Liver ◽

Liver Parenchyma ◽

Hepatoma Cell Lines ◽

Normal Rat ◽

Transformed Cell Lines

Abstract Esterases in nine rat hepatic and hepatoma-derived cell lines and normal rat liver homogenate were detected by polyacrylamide gel electrophoresis coupled with active staining with a-naphthyl acetate or butyrate as a substrate. The esterase band patterns of the non-cancerous and oncogene-transformed cell lines were alike, but different from those of hepatoma cell lines and normal rat liver homogenate. The former groups of cells might have completely lost the characteristics of rat liver parenchymal cells, or else they might have their origin at cells other than liver parenchyma. The esterase patterns of the hepatoma cell lines (e.g., McA-RH7777) and the normal rat liver highly resembled with each other, exemplifying the slight biochemical deviation of cancer from normal cells. However, two-dimensional electrophoretogram for the McA-RH7777 cell line showed a prominent esterase spot {p/ 6.0-Mr 110 kDa) that was lacking in the normal liver. This result indicates that there is invariably some change in esterase expression between the cancer cells and the normal liver cells

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Temozolomide sensitivity of malignant glioma cell lines – a systematic review assessing consistencies between in vitro studies

BMC Cancer ◽

10.1186/s12885-021-08972-5 ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Michael T. C. Poon ◽

Morgan Bruce ◽

Joanne E. Simpson ◽

Cathal J. Hannan ◽

Paul M. Brennan

Keyword(s):

Cell Viability ◽

Malignant Glioma ◽

Cell Line ◽

Cell Lines ◽

Glioma Cell ◽

Glioma Cell Line ◽

In Vitro Studies ◽

Experimental Conditions ◽

Glioma Cell Lines

Abstract Background Malignant glioma cell line models are integral to pre-clinical testing of novel potential therapies. Accurate prediction of likely efficacy in the clinic requires that these models are reliable and consistent. We assessed this by examining the reporting of experimental conditions and sensitivity to temozolomide in glioma cells lines. Methods We searched Medline and Embase (Jan 1994-Jan 2021) for studies evaluating the effect of temozolomide monotherapy on cell viability of at least one malignant glioma cell line. Key data items included type of cell lines, temozolomide exposure duration in hours (hr), and cell viability measure (IC50). Results We included 212 studies from 2789 non-duplicate records that reported 248 distinct cell lines. The commonest cell line was U87 (60.4%). Only 10.4% studies used a patient-derived cell line. The proportion of studies not reporting each experimental condition ranged from 8.0–27.4%, including base medium (8.0%), serum supplementation (9.9%) and number of replicates (27.4%). In studies reporting IC50, the median value for U87 at 24 h, 48 h and 72 h was 123.9 μM (IQR 75.3–277.7 μM), 223.1 μM (IQR 92.0–590.1 μM) and 230.0 μM (IQR 34.1–650.0 μM), respectively. The median IC50 at 72 h for patient-derived cell lines was 220 μM (IQR 81.1–800.0 μM). Conclusion Temozolomide sensitivity reported in comparable studies was not consistent between or within malignant glioma cell lines. Drug discovery science performed on these models cannot reliably inform clinical translation. A consensus model of reporting can maximise reproducibility and consistency among in vitro studies.

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In Vitro Cytotoxicity of Oleanolic/Ursolic Acids-Loaded in PLGA Nanoparticles in Different Cell Lines

Pharmaceutics ◽

10.3390/pharmaceutics11080362 ◽

2019 ◽

Vol 11 (8) ◽

pp. 362 ◽

Cited By ~ 20

Author(s):

Amélia M. Silva ◽

Helen L. Alvarado ◽

Guadalupe Abrego ◽

Carlos Martins-Gomes ◽

Maria L. Garduño-Ramirez ◽

...

Keyword(s):

Cell Viability ◽

Cell Line ◽

Cell Lines ◽

Specific Activity ◽

Polymeric Nanoparticles ◽

Hepatoma Cell ◽

Plga Nanoparticles ◽

Hepatoma Cell Line ◽

Human Hepatoma Cell

Oleanolic (OA) and ursolic (UA) acids are recognized triterpenoids with anti-cancer properties, showing cell-specific activity that can be enhanced when loaded into polymeric nanoparticles. The cytotoxic activity of OA and UA was assessed by Alamar Blue assay in three different cell lines, i.e., HepG2 (Human hepatoma cell line), Caco-2 (Human epithelial colorectal adenocarcinoma cell line) and Y-79 (Human retinoblastoma cell line). The natural and synthetic mixtures of these compounds were tested as free and loaded in polymeric nanoparticles in a concentration range from 2 to 32 µmol/L. The highest tested concentrations of the free triterpene mixtures produced statistically significant cell viability reduction in HepG2 and Caco-2 cells, compared to the control (untreated cells). When loaded in the developed PLGA nanoparticles, no differences were recorded for the tested concentrations in the same cell lines. However, in the Y-79 cell line, a decrease on cell viability was observed when testing the lowest concentration of both free triterpene mixtures, and after their loading into PLGA nanoparticles.

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Expression of Liver Functions in Imrnortalised Rot Hepotocyte Cell Lines

Human & Experimental Toxicology ◽

10.1177/096032719401300613 ◽

1994 ◽

Vol 13 (6) ◽

pp. 439-444 ◽

Cited By ~ 19

Author(s):

Caroline MacDonald ◽

Maureen Vass ◽

Brian Willett ◽

Alexander Scott ◽

Helen Grant

Keyword(s):

Cell Line ◽

Cell Lines ◽

Primary Cultures ◽

Rat Hepatocytes ◽

Hepatic Function ◽

Glutathione S Transferase ◽

Early Passage ◽

Toxicological Studies ◽

Well Differentiated

The differentiated hepatic function of two rat liver cell lines, P9 and SV40RH1, immortalised by transfection with SV40 DNA has been investigated in terms of the glutathione synthesis, and the activities of γ-glutamyltransferase, glutathione-S-transferase and UDP-glucuronosyltransferase. SV40RH1 is a highly differentiated cell line at early passage, but the expression of some aspects of its differentiated phenotype is unstable and some functions are lost by passage 12-13. P9 is a less-well differentiated cell line, with relatively stable expression of functions between passages 4 and 13. In terms of differentiated function both cell lines represent a marked improvement over primary cultures of rat hepatocytes which de-differentiate rapidly within 24-48 h in culture. This retention of liver function in proliferating cell lines offers the opportunity to use such cells in in vitro toxicological studies.

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