Liver cell lines for the study of hepatocyte functions and immunological response

Clayton, R. M.; Rinaldi, A.; Kandyba, Eve; Edward, Mike; Willberg, Christian; Klenerman, Paul; Patel, Arvind H.

doi:10.1111/j.1478-3231.2005.01017.x

Cited by 64 publications

(53 citation statements)

References 53 publications

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“…The HHL-5 cell line, derived from human liver cells, and retaining many phenotypic features of the liver, was the kind gift of Dr. A. Patel (Clayton et al, 2005).…”

Section: Cell Culture and Transfectionmentioning

confidence: 99%

Link between high‐affinity adenosine concentrative nucleoside transporter‐2 (CNT2) and energy metabolism in intestinal and liver parenchymal cells

Huber-Ruano

Pinilla-Macua

Torres

et al. 2010

Journal Cellular Physiology

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Concentrative nucleoside transporter 2 (CNT2) is a high-affinity adenosine transporter that may play physiological roles beyond nucleoside salvage. Previous reports relate CNT2 function to modulation of purinergic signaling and energy metabolism in intestinal and liver parenchymal cells (Duflot et al., 2004, Mol Cell Biol 24:2710-2719; Aymerich et al., 2006, J Cell Sci 119:1612-1621). In the present study, to further examine the link between CNT2 and energy metabolism, CNT2 protein partners were identified using the bacterial two-hybrid and GST pull-down approaches. The N-terminal segment of CNT2 was used as bait, since proteins lacking this domain display impaired plasma membrane insertion and intracellular retention. Glucose-regulated protein 58 (GRP58) was identified as a potential rCNT2 partner in pull-down experiments. Two-hybrid screening performed against a liver human cDNA library led to the identification of aldolase B as another hCNT2 partner. Aldolase B-RFP and endogenous GRP58 separately co-localized with CNT2 in HeLa cells transfected with YFPrCNT2. CNT2 interaction with GRP58 was validated using co-immunoprecipitation experiments. In HeLa cells, fluorescence resonance energy transfer (FRET) efficiency increased upon fructose addition, consistent with a transient interaction between aldolase B and the transporter. The physiological basis for in vivo interactions was derived from experiments in which GRP58 was inhibited or overexpressed and aldolase B activity stimulated towards glycolysis. GRP58 appeared to be a negative effector of CNT2 function, whereas aldolase B flux modulated CNT2 activity via a mechanism involving acquisition of higher affinity for its substrates. These findings support the theory that CNT2 plays roles other than salvage and establishes links with energy metabolism.

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“…The HHL-5 cell line, derived from human liver cells, and retaining many phenotypic features of the liver, was the kind gift of Dr. A. Patel (Clayton et al, 2005).…”

Section: Cell Culture and Transfectionmentioning

confidence: 99%

Link between high‐affinity adenosine concentrative nucleoside transporter‐2 (CNT2) and energy metabolism in intestinal and liver parenchymal cells

Huber-Ruano

Pinilla-Macua

Torres

et al. 2010

Journal Cellular Physiology

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show abstract

“…The expression of multiple other proteins, such as the Ntcp bile acid transporter, cytochrome P450 enzymes, also changes during the primary culture of hepatocytes and in established cell lines. [33][34][35] Thus, prior The established rat hepatoma cell line McARH was stably transfected with either a control plasmid or a plasmid to express the bile acid transporter Ntcp. As indicated, cells were pretreated for 6 hours with PBS vehicle (VEH) or PTX (300 ng/mL) followed by exposure to TCA (100 mol/L).…”

Section: Discussionmentioning

confidence: 99%

Conjugated bile acids promote ERK1/2 and AKT activation via a pertussis toxin-sensitive mechanism in murine and human hepatocytes

et al. 2005

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“…HeLa cells were maintained at 37°C in a humidified 5% CO 2 environment in Dulbecco's modified Eagle's medium supplemented with 10% (v/v) fetal bovine serum, 50 units/ml penicillin, 50 g/ml streptomycin, and 2 mM L-glutamine. The hepatocyte-derived cell line HHL5 was a generous gift from Dr. Arvin H. Patel (University of Glasgow, Glasgow, Scotland) (Clayton et al, 2005). HHL5 cells were maintained as described for HeLa cells, with the same culture medium but supplemented with 1% nonessential amino acids.…”

Section: Methodsmentioning

confidence: 99%

Role of the Transporter Regulator Protein (RS1) in the Modulation of Concentrative Nucleoside Transporters (CNTs) in Epithelia

et al. 2012

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SLC28 genes encode three plasma membrane transporter proteins, human concentrative nucleoside transporter (CNT)1, CNT2, and CNT3, all of which are implicated in the uptake of natural nucleosides and a variety of nucleoside analogs used in the chemotherapy of cancer and viral and inflammatory diseases. Mechanisms determining their trafficking toward the plasma membrane are not well known, although this might eventually become a target for therapeutic intervention. The transporter regulator RS1, which was initially identified as a short-term, post-transcriptional regulator of the high-affinity, Na ϩ -coupled, glucose transporter sodium-dependent glucose cotransporter 1, was evaluated in this study as a candidate for coordinate regulation of membrane insertion of human CNTtype proteins. With a combination of studies with mammalian cells, Xenopus laevis oocytes, and RS1-null mice, evidence that RS1 down-regulates the localization and activity at the plasma membrane of the three members of this protein family (CNT1, CNT2, and CNT3) is provided, which indicates the biochemical basis for coordinate regulation of nucleoside uptake ability in epithelia and probably in other RS1-expressing cell types.

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Liver cell lines for the study of hepatocyte functions and immunological response

Abstract: These HHLs retain primary hepatocyte phenotype and should be useful for investigating mechanisms of entry and replication of hepatotropic viruses, and should also be valuable in the study of hepatocyte biology and pathology.

Cited by 64 publications

References 53 publications

Link between high‐affinity adenosine concentrative nucleoside transporter‐2 (CNT2) and energy metabolism in intestinal and liver parenchymal cells

Link between high‐affinity adenosine concentrative nucleoside transporter‐2 (CNT2) and energy metabolism in intestinal and liver parenchymal cells

Conjugated bile acids promote ERK1/2 and AKT activation via a pertussis toxin-sensitive mechanism in murine and human hepatocytes

Role of the Transporter Regulator Protein (RS1) in the Modulation of Concentrative Nucleoside Transporters (CNTs) in Epithelia

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