Identification of a high molecular weight presumptive precursor to albumin mRNA in the nucleus of rat liver and hepatoma cell line H4AZC2.

The hormonal regulation of 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase gene expression was studied in the rat hepatoma cell line FAO-1. Both 6-phosphofructo-2-kinase and fructose-2,6-bisphosphatase activities were detected in FAO-1 cells, at 68% of the levels found in rat liver. Northern blot analysis showed that FAO-1 cells, like rat liver, contained a predominant species of bifunctional enzyme mRNA, which is 2.2 kb in size. A sensitive RNAase protection assay revealed the presence in FAO-1 cells of an additional mRNA species, which is generated when transcription is initiated from the skeletal muscle promoter of the rat liver/skeletal muscle gene. The liver/skeletal muscle mRNA ratio in FAO-1 cells was 10:1, which is similar to that observed in rat liver. In contrast, in another rat hepatoma cell line, FTO-2B, only the skeletal muscle mRNA was detected. Insulin and dexamethasone induced the liver bifunctional enzyme mRNA in FAO-1 cells by 2-4-fold and 10-20-fold respectively in a concentration- and time-dependent manner, and their effects were antagonized by cyclic AMP. Transcription of the gene in FAO-1 cells, measured by nuclear run-on assays, was also enhanced by dexamethasone and insulin. It is concluded that the FAO-1 cell line is similar to liver with respect to both the preferential use of the liver promoter of the gene and its regulation by hormones, and is therefore an excellent model for the study of the hepatic expression of this gene.

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Identification of a plasma membrane glutamine transporter from the rat hepatoma cell line H4-IIE-C3

Biochemical Journal ◽

10.1042/bj20020982 ◽

2002 ◽

Vol 368 (1) ◽

pp. 371-375 ◽

Cited By ~ 9

Author(s):

Matthew POLLARD ◽

David MEREDITH ◽

John D. McGIVAN

Keyword(s):

Amino Acid ◽

Cell Line ◽

Rat Liver ◽

Hepatoma Cell ◽

Single Amino Acid ◽

Hepatoma Cell Line ◽

Normal Rat ◽

Rat Hepatoma Cell Line ◽

Rat Hepatoma ◽

Glutamine Uptake

Glutamine is taken up into the rat hepatoma cell line H4-IIE-C3 by a Na+-dependent transport system which is specific for glutamine, alanine, serine, cysteine and asparagine and does not tolerate substitution of Na+ by Li+. Glutamine transport was relatively weakly inhibited by a 50-fold excess of leucine and was not inhibited by phenylalanine or N-methyl aminoisobutyrate. These general properties are characteristic of the recently identified ASCT/B0 family of transporters. Using a reverse transcriptase PCR-based homology cloning approach, we have characterized a cDNA for a novel member of this transporter family (H4-ASCT2) from H4-IIE-C3 cells. The cDNA encodes a 551-amino acid protein which exhibits similarities of between 75 and 85% with ASCT/B0 transporters previously cloned from other sources. When expressed in Xenopus oocytes, this transporter catalyses Na+-dependent glutamine uptake with characteristics very similar to those of glutamine uptake into the H4-IIE-C3 cells. This newly characterized transporter possesses a number of amino acid sequence differences from ASCT2 clones recently isolated from rat astroglial cells and from normal rat liver. In particular, the loop region between transmembrane helices 3 and 4 from H4-ASCT2 shares less than 60% sequence similarity with ASCT2 from rat liver; furthermore, there are some 25 single amino acid substitutions elsewhere in the H4-ASCT2 sequence compared with that from rat liver. Thus enhanced glutamine uptake in rat hepatoma cells is mediated by the expression of a novel ASCT/B0 transporter isoform rather than by increased expression of the ASCT2 mRNA found in normal rat liver.

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The aqueous extract from Artemisia capillaris Thunb. inhibits lipopolysaccharide-induced inflammatory response through preventing NF-κB activation in human hepatoma cell line and rat liver

International Journal of Molecular Medicine ◽

10.3892/ijmm.13.5.717 ◽

2004 ◽

Cited By ~ 1

Author(s):

Sang Hong ◽

Sang Seo ◽

Jun Lee ◽

Byung Choi

Keyword(s):

Inflammatory Response ◽

Cell Line ◽

Rat Liver ◽

Aqueous Extract ◽

Hepatoma Cell ◽

Hepatoma Cell Line ◽

Human Hepatoma ◽

Human Hepatoma Cell ◽

Human Hepatoma Cell Line ◽

Artemisia Capillaris

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The hormonal induction of gamma glutamyltransferase in rat liver and in a hepatoma cell line

Molecular and Cellular Biochemistry ◽

10.1007/bf00225247 ◽

1983 ◽

Vol 53-54 (1-2) ◽

Cited By ~ 11

Author(s):

Robert Barouki ◽

Marie-N�ble Chobert ◽

Jo�le Finidori ◽

Marie-Claude Billon ◽

Jacques Hanoune

Keyword(s):

Cell Line ◽

Rat Liver ◽

Hepatoma Cell ◽

Hepatoma Cell Line ◽

Gamma Glutamyltransferase ◽

Hormonal Induction

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Chromatin Low Molecular Weight RNA Components

Canadian Journal of Biochemistry ◽

10.1139/o73-112 ◽

1973 ◽

Vol 51 (6) ◽

pp. 903-912 ◽

Cited By ~ 10

Author(s):

John J. Monahan ◽

Ross H. Hall

Keyword(s):

Molecular Weight ◽

Cell Line ◽

Hepatoma Cell ◽

Chinese Hamster ◽

Low Molecular Weight ◽

Hepatoma Cell Line ◽

Kb Cells ◽

Chain Lengths ◽

Rat Hepatoma Cell Line ◽

Polynucleotide Chain

A low molecular weight RNA fraction has been isolated from chromatin of L-cells, KB cells, DON Chinese hamster cells, and a rat hepatoma cell line. This RNA fraction has been further fractionated into 11 subfractions which have widely different polynucleotide chain lengths. All subfractions are present in each of the cell lines, though the relative proportions of each differ from one cell line to another. The base composition and approximate s value of the 11 individual L-cell subfractions were determined. It is possible that each subfraction contains multiple RNA components.

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Comparison of the multiple deoxyribonucleic acid-dependent ribonucleic acid polymerase forms of whole rat liver and a minimal-deviation rat hepatoma cell line

Biochemical Journal ◽

10.1042/bj1260675 ◽

1972 ◽

Vol 126 (3) ◽

pp. 675-681 ◽

Cited By ~ 21

Author(s):

C. J. Chesterton ◽

S M Humphrey ◽

P H W Butterworth

Keyword(s):

Cell Line ◽

Rat Liver ◽

Animal Cell ◽

Hepatoma Cell ◽

Rna Polymerases ◽

Hepatoma Cell Line ◽

Enzyme Preparations ◽

Minimal Deviation ◽

Rat Hepatoma Cell Line ◽

Rat Hepatoma

To investigate the possibility that the pattern of multiple DNA-dependent RNA polymerases of an animal cell exerts a controlling influence on its nature, the activities of these enzymes were compared in differentiated rat liver and in a rapidly growing minimal-deviation rat hepatoma cell line by using established techniques of enzyme extraction, separation and determination. Relative to the DNA content of the tissues, RNA polymerase activities of forms AI, AII and B were approx. ninefold, twofold and twofold higher respectively in the cell line than in the liver. Tests indicated that these results could not be explained by differences in extraction efficiency or by the presence of unbound inhibitors or stimulators of polymerase activity in the final enzyme preparations. New forms of the enzyme were not detected in either tissue. The significance of these findings with respect to the possible role of multiple RNA polymerases in the control of cellular activities is discussed.

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Mechanisms of NADPH – cytochrome P450 oxidoreductase induction by dexamethasone in the H4IIE rat hepatoma cell line

Canadian Journal of Physiology and Pharmacology ◽

10.1139/cjpp-2019-0586 ◽

2020 ◽

Vol 98 (5) ◽

pp. 267-274

Author(s):

David S. Riddick ◽

Anne K. Mullen Grey

Keyword(s):

Cytochrome P450 ◽

Cell Line ◽

Rat Liver ◽

Hepatoma Cell ◽

Hepatoma Cell Line ◽

P450 Oxidoreductase ◽

Nadph Cytochrome ◽

Cytochrome P450 Oxidoreductase ◽

Rat Hepatoma Cell Line ◽

Rat Hepatoma

Expression of NADPH – cytochrome P450 oxidoreductase (POR), electron donor for microsomal P450s, is induced in rat liver by dexamethasone (DEX), an activator of the glucocorticoid receptor (GR) and the pregnane X receptor (PXR). DEX induction of POR in rat liver is primarily PXR-mediated, although GR may contribute to mRNA effects. We examined the role of GR and PXR in the DEX induction of POR mRNA and protein in the H4IIE rat hepatoma cell line. The DEX EC50 for a PXR target, CYP3A23, exceeded that for the GR targets tyrosine aminotransferase and PXR as well as POR itself. POR protein levels were induced 3- and 4-fold, respectively, by DEX concentrations activating GR selectively (100 nM) or both GR and PXR (10 μM). POR was induced by triamcinolone acetonide, a selective GR agonist, but not pregnenolone-16α-carbonitrile, a selective PXR agonist. POR induction was blocked by the GR antagonist RU486 but minimally influenced by the PXR antagonist FLB-12. The half-life for POR mRNA was prolonged by DEX at both 100 nM and 10 μM. GR is more important in DEX-induced POR expression in H4IIE cells compared to rat liver in vivo, calling into question the suitability of this cell model for mechanistic studies.

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Molecular and functional characterization of microsomal UDP-glucuronic acid uptake by members of the nucleotide sugar transporter (NST) family

Biochemical Journal ◽

10.1042/bj20060429 ◽

2006 ◽

Vol 400 (2) ◽

pp. 281-289 ◽

Cited By ~ 23

Author(s):

Tsutomu Kobayashi ◽

Judith E. Sleeman ◽

Michael W. H. Coughtrie ◽

Brian Burchell

Keyword(s):

Cell Line ◽

Rat Liver ◽

Glucuronic Acid ◽

Hepatoma Cell ◽

Functional Characterization ◽

Liver Microsomes ◽

Hepatoma Cell Line ◽

Acid Uptake ◽

Rat Liver Microsomes ◽

Nucleotide Sugar

Transport of the co-substrate UDPGA (UDP-glucuronic acid) into the lumen of the endoplasmic reticulum is an essential step in glucuronidation reactions due to the intraluminal location of the catalytic site of the enzyme UGT (UDP-glucuronosyltransferase). In the present study, we have characterized the function of several NSTs (nucleotide sugar transporters) and UGTs as potential carriers of UDPGA for glucuronidation reactions. UDPGlcNAc (UDP-N-acetylglucosamine)-dependent UDPGA uptake was found both in rat liver microsomes and in microsomes prepared from the rat hepatoma cell line H4IIE. The latency of UGT activity in microsomes derived from rat liver and V79 cells expressing UGT1A6 correlated well with mannose-6-phosphatase latency, confirming the UGT in the recombinant cells retained a physiology similar to rat liver microsomes. In the present study, four cDNAs coding for NSTs were obtained; two were previously reported (UGTrel1 and UGTrel7) and two newly identified (huYEA4 and huYEA4S). Localization of NSTs within the human genome sequence revealed that huYEA4S is an alternatively spliced form of huYEA4. All the cloned NSTs were stably expressed in V79 (Chinese hamster fibroblast) cells, and were able to transport UDPGA after preloading of isolated microsomal vesicles with UDPGlcNAc. The highest uptake was seen with UGTrel7, which displayed a Vmax approx. 1% of rat liver microsomes. Treatment of H4IIE cells with β-naphthoflavone induced UGT protein expression but did not affect the rate of UDPGA uptake. Furthermore, microsomes from UGT1-deficient Gunn rat liver showed UDPGA uptake similar to those from control rats. These data show that NSTs can act as UDPGA transporters for glucuronidation reactions, and indicate that UGTs of the 1A family do not function as UDPGA carriers in microsomes. The cell line H4IIE is a useful model for the study of UDPGA transporters for glucuronidation reactions.

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