The human hepatocyte cell lines IHH and HepaRG: models to study glucose, lipid and lipoprotein metabolism

Samanez, Carolina Huaman; Caron, Sandrine; Briand, Olivier; Dehondt, Hélène; Duplan, Isabelle; Kuipers, Folkert; Hennuyer, Nathalie; Clavey, Véronique; Staels, Bart

doi:10.3109/13813455.2012.683442

Cited by 50 publications

(40 citation statements)

References 36 publications

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“…Cell culture Immortalised human hepatocytes (IHHs) (a gift from B. Staels, the Pasteur Institute of Lille, University of Lille Nord de France, Lille, France [8]) were maintained in Complete William's E medium (Gibco, Paisley, UK) supplemented with bovine insulin (20 U/l; Sigma-Aldrich, St Louis, MO, USA) and dexamethasone (50 nmol/l; Sigma-Aldrich). HepG2 cells (hepatocellular carcinoma, human, American Type Culture Collection, Manassas, VA, USA) were maintained in DMEM (Lonza, Basel, Switzerland).…”

Section: Methodsmentioning

confidence: 99%

Protein kinase STK25 controls lipid partitioning in hepatocytes and correlates with liver fat content in humans

et al. 2015

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Aims/hypothesis Type 2 diabetes is closely associated with pathological lipid accumulation in the liver, which is suggested to actively contribute to the development of insulin resistance. We recently identified serine/threonine protein kinase 25 (STK25) as a regulator of liver steatosis, whole-body glucose tolerance and insulin sensitivity in a mouse model system. The aim of this study was to assess the role of STK25 in the control of lipid metabolism in human liver. Methods Intracellular fat deposition, lipid metabolism and insulin sensitivity were studied in immortalised human hepatocytes (IHHs) and HepG2 hepatocellular carcinoma cells in which STK25 was overexpressed or knocked down by small interfering RNA. The association between STK25 mRNA expression in human liver biopsies and hepatic fat content was analysed.Results Overexpression of STK25 in IHH and HepG2 cells enhanced lipid deposition by suppressing β-oxidation and triacylglycerol (TAG) secretion, while increasing lipid synthesis. Conversely, knockdown of STK25 attenuated lipid accumulation by stimulating β-oxidation and TAG secretion, while inhibiting lipid synthesis. Furthermore, TAG hydrolase activity was repressed in hepatocytes overexpressing STK25 and reciprocally increased in cells with STK25 knockdown. Insulin sensitivity was reduced in STK25-overexpressing cells and enhanced in STK25-deficient hepatocytes. We also found a statistically significant positive correlation between STK25 mRNA expression in human liver biopsies and hepatic fat content. Conclusions/interpretation Our data suggest that STK25 regulates lipid partitioning in human liver cells by controlling TAG synthesis as well as lipolytic activity and thereby NEFA release from lipid droplets for β-oxidation and TAG secretion. Our findings highlight STK25 as a potential drug target for the prevention and treatment of type 2 diabetes.

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Section: Methodsmentioning

confidence: 99%

Protein kinase STK25 controls lipid partitioning in hepatocytes and correlates with liver fat content in humans

et al. 2015

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“…Total RNA was prepared from IHH cells stimulated by low and high glucose concentrations, as previously described (31), in the presence of DMSO or GW4064 (5 M) for 24 h. mRNA quality was analyzed on a Bioanalyzer 2100 (Agilent Technologies), and only RNA preparations with high integrity (RNA integrity number [RIN] of Ͼ9) were used. RNA was amplified and labeled using the quick amp labeling kit (Agilent Technologies) and hybridized to Agilent whole human genome GE 4x44K microarrays (Agilent Technologies).…”

Section: Methodsmentioning

confidence: 99%

“…IHH and HepaRG cells were cultured and stimulated with 11 mM and 25 mM glucose, respectively, as described previously (31,33,34). For some experiments, as indicated below, IHH cells were pretreated for 1 h with trichostatin A (TSA) (300 g/ml) before incubation with glucose.…”

Section: Methodsmentioning

confidence: 99%

“…IHH and HepaRG cells were transfected as previously described (31,35) with the following small interfering RNAs (siRNAs): OnTargetPlus SmartPools, scramble control (product no. D-001810; Dharmacon, Lafayette, CO), FXR (product no.…”

Section: Methodsmentioning

confidence: 99%

“…Furthermore, this regulation is conserved in immortalized human hepatocytes (IHH), which display glucose and insulin responsiveness (31). Investigation of the molecular mechanisms by which FXR inhibits the response to glucose shows that FXR inhibits glucose-induced L-PK expression by a transrepressive mechanism involving the release of the transcription factor ChREBP and the recruitment of the transcriptional corepressor SMRT to the ChORE of the L-PK promoter.…”

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confidence: 99%

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Farnesoid X Receptor Inhibits the Transcriptional Activity of Carbohydrate Response Element Binding Protein in Human Hepatocytes

Caron

Samanez

Dehondt

et al. 2013

Molecular and Cellular Biology

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120

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Glucose sensing O-GlcNAcylation pathway regulates the nuclear bile acid receptor farnesoid X receptor (FXR)

et al. 2014

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Bile acid metabolism is intimately linked to the control of energy homeostasis and glucose and lipid metabolism. The nuclear receptor farnesoid X receptor (FXR) plays a major role in the enterohepatic cycling of bile acids, but the impact of nutrients on bile acid homeostasis is poorly characterized. Metabolically active hepatocytes cope with increases in intracellular glucose concentrations by directing glucose into storage (glycogen) or oxidation (glycolysis) pathways, as well as to the pentose phosphate shunt and the hexosamine biosynthetic pathway. Here we studied whether the glucose nonoxidative hexosamine biosynthetic pathway modulates FXR activity. Our results show that FXR interacts with and is OGlcNAcylated by O-GlcNAc transferase in its N-terminal AF1 domain. Increased FXR OGlcNAcylation enhances FXR gene expression and protein stability in a cell type-specific manner. High glucose concentrations increased FXR O-GlcNAcylation, hence its protein stability and transcriptional activity by inactivating corepressor complexes, which associate in a ligand-dependent manner with FXR, and increased FXR binding to chromatin. Finally, in vivo fasting-refeeding experiments show that FXR undergoes O-GlcNAcylation in fed conditions associated with increased direct FXR target gene expression and decreased liver bile acid content. Conclusion: FXR activity is regulated by glucose fluxes in hepatocytes through a direct posttranslational modification catalyzed by the glucose-sensing hexosamine biosynthetic pathway. (HEPATOLOGY 2014;59:2023-2034 See Editorial on Page 1665 T he liver plays an important role in energy homeostasis by constantly tuning metabolic networks in response to varying nutrient fluxes. Marked variations in blood glucose concentrations occur daily, which are rapidly compensated by an integrated response of several tissues, such as the liver, which reacts by adjusting the relative activity of gluconeogenic, glycogenogenic and glycolytic pathways, hence contributing to the maintenance of glucose homeostasis. Hepatocytes also direct glucose into the pentose phosphate shunt and the hexosamine biosynthetic pathways (HBP), the latter consuming up to 5% of the cellular glucose. HBP generates uridine diphosphate N-acetyl-glucosamine (UDP-GlcNAc) from glucose, glutamine, acetyl-coenzyme A, uridine, and adenosine triphosphate (ATP). UDP-GlcNAc synthesis is catalyzed by Abbreviations: BA, bile acid; DON,6

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The human hepatocyte cell lines IHH and HepaRG: models to study glucose, lipid and lipoprotein metabolism

Cited by 50 publications

References 36 publications

Protein kinase STK25 controls lipid partitioning in hepatocytes and correlates with liver fat content in humans

Protein kinase STK25 controls lipid partitioning in hepatocytes and correlates with liver fat content in humans

Farnesoid X Receptor Inhibits the Transcriptional Activity of Carbohydrate Response Element Binding Protein in Human Hepatocytes

Glucose sensing O-GlcNAcylation pathway regulates the nuclear bile acid receptor farnesoid X receptor (FXR)

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