scholarly journals RORα phosphorylation by casein kinase 1α as glucose signal to regulate estrogen sulfation in human liver cells

2020 ◽  
Vol 477 (18) ◽  
pp. 3583-3598 ◽  
Author(s):  
Hao Hu ◽  
Masahiko Negishi
Keyword(s):  
Nuclear Receptor ◽  
High Glucose ◽  
Hepg2 Cells ◽  
Human Liver ◽  
Nuclear Protein ◽  
Casein Kinase ◽  
Promoter Activation ◽  
Distal Enhancer ◽  
Estrogen Sulfotransferase

Estrogen sulfotransferase (SULT1E1) metabolically inactivates estrogen and SULT1E1 expression is tightly regulated by multiple nuclear receptors. Human fetal, but not adult, livers express appreciable amounts of SULT1E1 protein, which is mimicked in human hepatoma-derived HepG2 cells cultured in high glucose (450 mg/dl) medium. Here, we have investigated this glucose signal that leads to phosphorylation of nuclear receptor RORα (NR1F1) at Ser100 and the transcription mechanism by which phosphorylated RORα transduces this signal to nuclear receptor HNF4α, activating the SULT1E1 promoter. The promoter is repressed by non-phosphorylated RORα which binds a distal enhancer (−943/−922 bp) and interacts with and represses HNF4α-mediated transcription. In response to high glucose, RORα becomes phosphorylated at Ser100 and reverses its repression of HNF4α promoter activation. Moreover, the casein kinase CK1α, which is identified in an enhancer-bound nuclear protein complex, phosphorylates Ser100 in in vitro kinase assays. During these dynamic processes, both RORα and HNF4α remain on the enhancer. Thus, RORα utilizes phosphorylation to integrate HNF4α and transduces the glucose signal to regulate the SULT1E1 gene in HepG2 cells and this phosphorylation-mediated mechanism may also regulate SULT1E1 expressions in the human liver.

AAnti-Inflammatory Effects of Plasma Circulating Exosomes Obtained from Normal-Weight and Obese Subjects on Hepatocytes

2020 ◽  
Vol 20 ◽  
Author(s):  
Reza Afrisham ◽  
Sahar Sadegh-Nejadi ◽  
Reza Meshkani ◽  
Solaleh Emamgholipour ◽  
Molood Bagherieh ◽  
...  
Keyword(s):  
Inflammatory Cytokines ◽  
Hepg2 Cells ◽  
Human Liver ◽  
Liver Cells ◽  
Normal Weight ◽  
Protein Levels ◽  
Human Liver Cells ◽  
Tnf Α ◽  
Anti Inflammatory

Introduction: Obesity is a disorder with low-grade chronic inflammation that plays a key role in the hepatic inflammation and steatosis. Moreover, there are studies to support the role of exosomes in the cellular communications, the regulation of metabolic homeostasis and immunomodulatory activity. Accordingly, we aimed to evaluate the influence of plasma circulating exosomes derived from females with normal-weight and obesity on the secretion of inflammatory cytokines in human liver cells. Methods: Plasma circulating exosomes were isolated from four normal (N-Exo) and four obese (O-Exo) women. The exosomes were characterized and approved for CD63 expression (common exosomal protein marker) and morphology/size using the western blot and TEM methods, respectively. The exosomes were used for stimulation of HepG2 cells in vitro. After 24 h incubation, the protein levels of TNF-α,IL-6, and IL-1β were measured in the culture supernatant of HepG2 cells using the ELISA kit. Results: The protein levels of IL-6 and TNF-α in the cells treated with O-Exo and N-Exo reduced significantly in comparison with control group (P=0.039 and P<0.001 respectively), while significance differences were not found between normal and obese groups (P=0.808, and P=0.978 respectively). However, no significant differences were found between three groups in term of IL-1β levels (P=0.069). Based on the correlation analysis, the protein levels of IL-6 were positively correlated with TNF-α (r 0.978, P<0.001). Conclusion: These findings suggest that plasma circulating exosomes have probably anti-inflammatory properties independently from body mass index and may decrease the secretion of inflammatory cytokines in liver. However, further investigations in vitro and in vivo are needed to address the anti-inflammatory function of N-Exo and O-Exo in human liver cells and/or other cells.

Development of in vitro model of insulin receptor cleavage induced by high glucose in HepG2 cells

2014 ◽  
Vol 445 (1) ◽  
pp. 236-243 ◽  
Author(s):  
Tomoyuki Yuasa ◽  
Kikuko Amo ◽  
Shuhei Ishikura ◽  
Hisao Nagaya ◽  
Keiji Uchiyama ◽  
...  
Keyword(s):  
Insulin Receptor ◽  
High Glucose ◽  
Hepg2 Cells ◽  
In Vitro Model ◽  
Vitro Model

Metabolism of Ebracteolata Compound B Studied In Vitro with Human Liver Microsomes, HepG2 Cells, and Recombinant Human Enzymes

2010 ◽  
Vol 38 (12) ◽  
pp. 2157-2165 ◽  
Author(s):  
Xia Zhang ◽  
Yan Yao ◽  
Yan Lou ◽  
Huidi Jiang ◽  
Xiaowen Wang ◽  
...  
Keyword(s):  
Hepg2 Cells ◽  
Human Liver ◽  
Liver Microsomes ◽  
Human Liver Microsomes

scholarly journals Comparing in vitro human liver models to in vivo human liver using RNA-Seq

2020 ◽  
Author(s):  
Rajinder Gupta ◽  
Yannick Schrooders ◽  
Duncan Hauser ◽  
Marcel van Herwijnen ◽  
Wiebke Albrecht ◽  
...  
Keyword(s):  
Liver Cell ◽  
Liver Tissue ◽  
Hepg2 Cells ◽  
Human Liver ◽  
Rna Seq ◽  
Cell Models ◽  
Human Liver Tissue ◽  
Human Liver Cell

Abstract The liver plays an important role in xenobiotic metabolism and represents a primary target for toxic substances. Many different in vitro cell models have been developed in the past decades. In this study, we used RNA-sequencing (RNA-Seq) to analyze the following human in vitro liver cell models in comparison to human liver tissue: cancer-derived cell lines (HepG2, HepaRG 3D), induced pluripotent stem cell-derived hepatocyte-like cells (iPSC-HLCs), cancerous human liver-derived assays (hPCLiS, human precision cut liver slices), non-cancerous human liver-derived assays (PHH, primary human hepatocytes) and 3D liver microtissues. First, using CellNet, we analyzed whether these liver in vitro cell models were indeed classified as liver, based on their baseline expression profile and gene regulatory networks (GRN). More comprehensive analyses using non-differentially expressed genes (non-DEGs) and differential transcript usage (DTU) were applied to assess the coverage for important liver pathways. Through different analyses, we noticed that 3D liver microtissues exhibited a high similarity with in vivo liver, in terms of CellNet (C/T score: 0.98), non-DEGs (10,363) and pathway coverage (highest for 19 out of 20 liver specific pathways shown) at the beginning of the incubation period (0 h) followed by a decrease during long-term incubation for 168 and 336 h. PHH also showed a high degree of similarity with human liver tissue and allowed stable conditions for a short-term cultivation period of 24 h. Using the same metrics, HepG2 cells illustrated the lowest similarity (C/T: 0.51, non-DEGs: 5623, and pathways coverage: least for 7 out of 20) with human liver tissue. The HepG2 are widely used in hepatotoxicity studies, however, due to their lower similarity, they should be used with caution. HepaRG models, iPSC-HLCs, and hPCLiS ranged clearly behind microtissues and PHH but showed higher similarity to human liver tissue than HepG2 cells. In conclusion, this study offers a resource of RNA-Seq data of several biological replicates of human liver cell models in vitro compared to human liver tissue.

scholarly journals Characterization of lipid metabolism in a novel immortalized human hepatocyte cell line

2015 ◽  
Vol 309 (6) ◽  
pp. E511-E522 ◽  
Author(s):  
Charlotte J. Green ◽  
Deborah Johnson ◽  
Harsh D. Amin ◽  
Pamela Sivathondan ◽  
Michael A. Silva ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
Cell Line ◽  
High Glucose ◽  
Hepg2 Cells ◽  
Human Liver ◽  
Liver Fat ◽  
Primary Hepatocytes ◽  
Human Liver Cell ◽  
Intracellular Triglyceride

The development of hepatocyte cell models that represent fatty acid partitioning within the human liver would be beneficial for the study of the development and progression of nonalcoholic fatty liver disease (NAFLD). We sought to develop and characterize a novel human liver cell line (LIV0APOLY) to establish a model of lipid accumulation using a physiological mixture of fatty acids under low- and high-glucose conditions. LIV0APOLY cells were compared with a well-established cell line (HepG2) and, where possible, primary human hepatocytes. LIV0APOLY cells were found to proliferate and express some mature liver markers and were wild type for the PNPLA3 (rs738409) gene, whereas HepG2 cells carried the Ile148Met variant that is positively associated with liver fat content. Intracellular triglyceride content was higher in HepG2 than in LIV0APOLY cells; exposure to high glucose and/or exogenous fatty acids increased intracellular triglyceride in both cell lines. Triglyceride concentrations in media were higher from LIV0APOLY compared with HepG2 cells. Culturing LIV0APOLY cells in high glucose increased a marker of endoplasmic reticulum stress and attenuated insulin-stimulated Akt phosphorylation whereas low glucose and exogenous fatty acids increased AMPK phosphorylation. Although LIV0APOLY cells and primary hepatocytes stored similar amounts of exogenous fatty acids as triglyceride, more exogenous fatty acids were partitioned toward oxidation in the LIV0APOLY cells than in primary hepatocytes. LIV0APOLY cells offer the potential to be a renewable cellular model for studying the effects of exogenous metabolic substrates on fatty acid partitioning; however, their usefulness as a model of lipoprotein metabolism needs to be further explored.

scholarly journals Organophosphorus Flame Retardant TDCPP Displays Genotoxic and Carcinogenic Risks in Human Liver Cells

Cells ◽  
2022 ◽  
Vol 11 (2) ◽  
pp. 195
Author(s):  
Quaiser Saquib ◽  
Abdullah M. Al-Salem ◽  
Maqsood A. Siddiqui ◽  
Sabiha M. Ansari ◽  
Xiaowei Zhang ◽  
...  
Keyword(s):  
Flame Retardant ◽  
Hepg2 Cells ◽  
Human Liver ◽  
Human Cancer ◽  
Liver Cells ◽  
Consumer Products ◽  
In Vivo Test ◽  
Human Liver Cells

Tris(1,3-Dichloro-2-propyl)phosphate (TDCPP) is an organophosphorus flame retardant (OPFR) widely used in a variety of consumer products (plastics, furniture, paints, foams, and electronics). Scientific evidence has affirmed the toxicological effects of TDCPP in in vitro and in vivo test models; however, its genotoxicity and carcinogenic effects in human cells are still obscure. Herein, we present genotoxic and carcinogenic properties of TDCPP in human liver cells (HepG2). 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) and neutral red uptake (NRU) assays demonstrated survival reduction in HepG2 cells after 3 days of exposure at higher concentrations (100–400 μM) of TDCPP. Comet assay and flow cytometric cell cycle experiments showed DNA damage and apoptosis in HepG2 cells after 3 days of TDCPP exposure. TDCPP treatment incremented the intracellular reactive oxygen species (ROS), nitric oxide (NO), Ca2+ influx, and esterase level in exposed cells. HepG2 mitochondrial membrane potential (ΔΨm) significantly declined and cytoplasmic localization of P53, caspase 3, and caspase 9 increased after TDCPP exposure. qPCR array quantification of the human cancer pathway revealed the upregulation of 11 genes and downregulation of two genes in TDCPP-exposed HepG2 cells. Overall, this is the first study to explicitly validate the fact that TDCPP bears the genotoxic, hepatotoxic, and carcinogenic potential, which may jeopardize human health.

scholarly journals Up-regulation of CXCL8 expression is associated with a poor prognosis and enhances tumor cell malignant behaviors in liver cancer

2020 ◽  
Vol 40 (8) ◽  
Author(s):  
Song Yang ◽  
Han Wang ◽  
Chundi Qin ◽  
Hongmei Sun ◽  
Yupeng Han
Keyword(s):  
Liver Cancer ◽  
Hepg2 Cells ◽  
Human Liver ◽  
Clinical Stage ◽  
Critical Role ◽  
Human Liver Tissue ◽  
Underlying Mechanisms ◽  
And Migration ◽  
High Level

Abstract CXCL8, a member of CXC chemokines, was constitutively expressed in many types of human cancers, and its overexpression has been shown to play a critical role in promoting tumorigenesis. The purpose of the present study was to determine CXCL8 expression in a commercial human liver tissue microarray, and elucidate the effects and underlying mechanisms by which CXCL8 is involved in the malignant progression of human liver cancer. Our data showed that high level expression of CXCL8 in tissues with liver cancer was identified as compared with non-cancer tissues, and its up-regulation was closely associated with clinical stage and tumor infiltration. In vitro, exogenous CXCL8 at concentrations of 10, 20 or 40 ng/ml obviously stimulated the proliferation abilities of HepG2 cells. Coupled with this, 10, 20 or 40 ng/ml of exogenous CXCL8 also triggered a significant elevation in HepG2 cells migration. Additionally, overexpression of CXCL8 in HepG2 cells also resulted in increased cell proliferation and migration capacities. Finally, Western blotting analysis showed that overexpression of CXCL8 increased the expression of ERK, p-ERK and survivin, decreased the expression of caspase-3 and BAX at protein level.

Speciation studies of vanadium in human liver (HepG2) cells after in vitro exposure to bis(maltolato)oxovanadium(iv) using HPLC online with elemental and molecular mass spectrometry

Metallomics ◽  
2013 ◽  
Vol 5 (12) ◽  
pp. 1685 ◽  
Author(s):  
Volker Nischwitz ◽  
Julie T. Davies ◽  
Damian Marshall ◽  
Macarena González ◽  
Jose Luis Gómez Ariza ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Molecular Mass ◽  
Hepg2 Cells ◽  
Human Liver ◽  
In Vitro Exposure ◽  
Molecular Mass Spectrometry
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