Differential signatures of protein glycosylation and phosphorylation in human Chang liver cells induced by TCDD treatment

BackgroundIn vitro cell systems together with omics methods represent promising alternatives to conventional animal models for toxicity testing. Transcriptomic and proteomic approaches have been widely applied in vitro but relatively few studies have used metabolomics. Therefore, the goal of the present study was to develop an untargeted methodology for performing reproducible metabolomics on in vitro systems. The human liver cell line HepG2, and the well-known hepatotoxic and non-genotoxic carcinogen 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), were used as the in vitro model system and model toxicant, respectively.ResultsThe study focused on the analysis of intracellular metabolites using NMR, LC-MS and GC-MS, with emphasis on the reproducibility and repeatability of the data. State of the art pre-processing and alignment tools and multivariate statistics were used to detect significantly altered levels of metabolites after exposing HepG2 cells to TCDD. Several metabolites identified using databases, literature and LC-nanomate-Orbitrap analysis were affected by the treatment. The observed changes in metabolite levels are discussed in relation to the reported effects of TCDD.ConclusionsUntargeted profiling of the polar and apolar metabolites of in vitro cultured HepG2 cells is a valid approach to studying the effects of TCDD on the cell metabolome. The approach described in this research demonstrates that highly reproducible experiments and correct normalization of the datasets are essential for obtaining reliable results. The effects of TCDD on HepG2 cells reported herein are in agreement with previous studies and serve to validate the procedures used in the present work.

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“…Altered expression levels of glycosylated protein have been described after TCDD exposure in vitro [76]. …”

Section: Discussionmentioning

confidence: 99%

An untargeted multi-technique metabolomics approach to studying intracellular metabolites of HepG2 cells exposed to 2,3,7,8-tetrachlorodibenzo-p-dioxin

et al. 2011

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“…The specific mechanisms by which TCDD exerts these effects, either via the AHR or independently, are unknown. However, recent evidence suggests that TCDD may alter the function of cytoskeletal proteins by changing posttranslational modifications [36]. In particular, b-actin was differently glycosylated, and the microtubule-associated protein 1S (MAP1S) and actin-related protein 1 (ARP1) homolog A showed enhanced tyrosine phosphorylation when cells were exposed to TCDD.…”

Section: Discussionmentioning

confidence: 99%

The Environmental Toxicant 2,3,7,8-Tetrachlorodibenzo-p-Dioxin Disturbs the Establishment and Maintenance of Cell Polarity in Preimplantation Rat Embryos1

Hutt

Shi

Petroff

et al. 2010

Biology of Reproduction

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Maternal exposure to the environmental toxicant 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) induces a variety of defects in compaction-stage embryos, including monopolar spindle formation, errors in chromosome segregation, and fragmentation resulting from aberrant cytokinesis. In this study, we investigated the possibility that a failure in centrosome duplication, separation, or positioning within blastomeres might underlie the observed effects of TCDD on early embryos. The subcellular localization of the centrosomal marker TUBG1 was analyzed in preimplantation embryos collected from female rats exposed to either chronic (50 ng kg(-1) wk(-1) for 3 wk) or acute (50 ng/kg or 1 microg/kg at proestrus) doses of TCDD. In treated embryos, interphase TUBG1 foci were more abundant and cortically displaced when compared to those in controls. At prophase, some blastomeres exhibited a single large perinuclear TUBG1 aggregate, suggesting a failure in centrosome duplication or separation. Furthermore, the presence of monopolar spindles at metaphase was confirmed by the localization of TUBG1 to the single spindle pole. Therefore, the misregulation of centrosome number and localization, as indicated by TUBG1 staining, may contribute to errors in chromosome segregation and cytokinesis in embryos following maternal TCDD exposure.

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“…The differently expression of proteins are expected to provide insights into the various molecular mechanisms. Previously, the major proteomic studies are emphasized that the aspect of liver lesion is underlying TCDD-induced toxicity according to the modulation of drug-metabolizing enzymes [29][30][31]. Although the lung lesion and cancer are considered to associate with TCDD exposure, the systemic proteomic analysis has not been reported.…”

Section: Introductionmentioning

confidence: 99%

Proteomics Investigation Reveals Apoptosis-Associated Proteins in Aryl Hydrocarbon Receptor-Deficient Human Lung Cells Treated with 2,3,7,8-Tetrachlorobenzo-p-dioxin

Hsiao

Yin

Lin

et al. 2012

JPB

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Differential signatures of protein glycosylation and phosphorylation in human Chang liver cells induced by TCDD treatment

Cited by 12 publications

References 41 publications

An untargeted multi-technique metabolomics approach to studying intracellular metabolites of HepG2 cells exposed to 2,3,7,8-tetrachlorodibenzo-p-dioxin

An untargeted multi-technique metabolomics approach to studying intracellular metabolites of HepG2 cells exposed to 2,3,7,8-tetrachlorodibenzo-p-dioxin

The Environmental Toxicant 2,3,7,8-Tetrachlorodibenzo-p-Dioxin Disturbs the Establishment and Maintenance of Cell Polarity in Preimplantation Rat Embryos1

Proteomics Investigation Reveals Apoptosis-Associated Proteins in Aryl Hydrocarbon Receptor-Deficient Human Lung Cells Treated with 2,3,7,8-Tetrachlorobenzo-p-dioxin

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