Use of human-derived liver cell lines for the detection of environmental and dietary genotoxicants; current state of knowledge

Knasmüller, Siegfried; Mersch-Sundermann, Volker; Kevekordes, Sebastian; Darroudi, F.; Huber, Wolfgang; Hoelzl, Christine; Bichler, Julia; Majer, Bernhard

doi:10.1016/j.tox.2004.02.008

Cited by 315 publications

(147 citation statements)

References 66 publications

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“…The hepatoblastoma cell line HepG2 was employed in the screen, because it retains better hepatic characteristics, has been widely used for cytotoxicity studies [21], and shares some important genetic alterations with hepatocellular carcinoma (HCC) [22]. In addition, doxorubicin, a widely used front-line chemotherapeutic drug, was used in the screen.…”

Section: Identification Of Mirnas That Enhance Drug Sensitivity By Fumentioning

confidence: 99%

miR-27b synergizes with anticancer drugs via p53 activation and CYP1B1 suppression

Zhang

et al. 2015

Cell Res

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Liver and kidney cancers are notorious for drug resistance. Due to the complexity, redundancy and interpatient heterogeneity of resistance mechanisms, most efforts targeting a single pathway were unsuccessful. Novel personalized therapies targeting multiple essential drug resistance pathways in parallel hold a promise for future cancer treatment. Exploiting the multitarget characteristic of microRNAs (miRNAs), we developed a new therapeutic strategy by the combinational use of miRNA and anticancer drugs to increase drug response. By a systems approach, we identified that miR-27b, a miRNA deleted in liver and kidney cancers, sensitizes cancer cells to a broad spectrum of anticancer drugs in vitro and in vivo. Functionally, miR-27b enhances drug response by activating p53-dependent apoptosis and reducing CYP1B1-mediated drug detoxification. Notably, miR-27b promotes drug response specifically in patients carrying p53-wild-type or CYP1B1-high signature. Together, we propose that miR-27b synergizes with anticancer drugs in a defined subgroup of liver and kidney cancer patients.

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Section: Identification Of Mirnas That Enhance Drug Sensitivity By Fumentioning

confidence: 99%

miR-27b synergizes with anticancer drugs via p53 activation and CYP1B1 suppression

Zhang

et al. 2015

Cell Res

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“…In addition to L5178Y cells, CHO [24][25][26], V79 [27], HepG2 [28], TK6 [29], and other cell lines have various advantageous characteristics. For instance, preliminary work with the attachment cell lines CHO-K1 and HepG2 indicate that the EMA dye and subsequent lysis solutions described herein can be directly applied to cells that are affixed to growth vessels, thereby liberating nuclei and MN without the need for trypsinization.…”

Section: Compatibility With Other Cell Linesmentioning

confidence: 99%

Interlaboratory evaluation of a flow cytometric, high content in vitro micronucleus assay

Bryce

Avlasevich

Bemis

et al. 2008

Mutation Research/Genetic Toxicology and Environmental Mutagene

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An international, multi-lab trial was conducted to evaluate a flow cytometry-based method for scoring micronuclei in mouse lymphoma L5178Y cells [Avlasevich et al., Environ. Molec. Mutagen. 47 (2006) [56][57][58][59][60][61][62][63][64][65][66]. A reference laboratory investigated the potential of six chemicals to induce micronuclei-the genotoxicants mitomycin C, etoposide, and vinblastine, and the non-genotoxicants sucrose, staurosporine, or dexamethasone. The latter two non-genotoxicants were selected as extreme challenges to the assay because of their potent apoptogenic activity. Three collaborating laboratories were supplied with prototype In Vitro MicroFlow ™ kits, and each was assigned one genotoxicant and one non-genotoxicant. Cells were treated continuously for 24 hrs over a range of concentrations up to 5 mg/ml, or overtly cytotoxic concentrations. Micronuclei were scored via standard microscopy and flow cytometry. In addition to enumerating micronucleus frequencies, a cytotoxicity measurement that is simultaneously acquired with the flow cytometric micronucleus scoring procedure was evaluated (Flow-NBR). With this method, latex particles served as counting beads, and facilitated relative survival measurements that exclude the presence of dead/dying cells. For comparison purposes, additional cytotoxicity endpoints were measured, including several that are based on cell number, and others that reflect compromised membrane integrity, including dye permeability and/or phospholipid distribution. Key findings for this set of compounds include the following: (1) significant discrepancies in top concentration selection were found when cytotoxicity measurements were based on different methods, with the Flow-NBR approach tending to be the most sensitive, (2) both microscopy-and flow cytometry-based scoring methods detected concentrationdependent micronucleus formation for the three genotoxic agents studied, with good agreement between the reference laboratory and the collaborating laboratories, and (3) whereas flow cytometric analyses showed no significant increases for the non-genotoxicants when top concentration selection was based on Flow-NBR, significantly elevated micronucleus frequencies were observed for concentrations that were chosen based on less-sensitive cytotoxicity assays. Collectively, these results indicate that rapid assessment of genotoxicity can be accomplished with a relatively simple Publisher's Disclaimer: This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final citable form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. NIH Public Access

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“…19,20 Cell lines of hepatic origin, such as HepG2 cells, have been previously adapted to HTS formats 21,22 and utilized to assess hepatotoxicity. 19,20,23 However, HepG2 cells lack the full expression of hepatocyte proteins, such as phase I and phase II metabolizing enzymes and transporters, and thus may not correlate to in vivo hepatotoxicity. [24][25][26] As an alternative to HepG2 cells, primary human hepatocytes represent the best predictive in vitro model to determine liver function for metabolism, 27,28 drug-drug interactions, 29,30 and potential hepatotoxicity of compounds.…”

Section: Introductionmentioning

confidence: 99%

Assessment of Compound Hepatotoxicity Using Human Plateable Cryopreserved Hepatocytes in a 1536-Well-Plate Format

Moeller¹,

Shukla

Xia

2012

ASSAY and Drug Development Technologies

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Hepatotoxicity is a major concern for both drug development and toxicological evaluation of environmental chemicals. The assessment of compound-induced hepatotoxicity has traditionally relied on in vivo testing; however, it is being replaced by human in vitro models due to an emphasis on the reduction of animal testing and species-specific differences. Since most cell lines and hybridomas lack the full complement of enzymes at physiological levels found in the liver, primary hepatocytes are the gold standard to study liver toxicities in vitro due to the retention of most of their in vivo activities. Here, we optimized a cell viability assay using plateable cryopreserved human hepatocytes in a 1536-well-plate format. The assay was validated by deriving inhibitory concentration at 50% values for 12 known compounds, including tamoxifen, staurosporine, and phenylmercuric acetate, with regard to hepatotoxicity and general cytotoxicity using multiple hepatocyte donors. The assay performed well, and the cytotoxicity of these compounds was confirmed in comparison to HepG2 cells. This is the first study to report the reliability of using plateable cryopreserved human hepatocytes for cytotoxicity studies in a 1536-well-plate format. These results suggest that plateable cryopreserved human hepatocytes can be scaled up for screening a large compound library and may be amenable to other hepatocytic assays such as metabolic or drug safety studies.

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Use of human-derived liver cell lines for the detection of environmental and dietary genotoxicants; current state of knowledge

Cited by 315 publications

References 66 publications

miR-27b synergizes with anticancer drugs via p53 activation and CYP1B1 suppression

miR-27b synergizes with anticancer drugs via p53 activation and CYP1B1 suppression

Interlaboratory evaluation of a flow cytometric, high content in vitro micronucleus assay

Assessment of Compound Hepatotoxicity Using Human Plateable Cryopreserved Hepatocytes in a 1536-Well-Plate Format

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