scholarly journals Transferrin receptor 2 is crucial for iron sensing in human hepatocytes

2010 ◽  
Vol 299 (3) ◽  
pp. G778-G783 ◽  
Author(s):  
Chiara Rapisarda ◽  
Juliana Puppi ◽  
Robin D. Hughes ◽  
Anil Dhawan ◽  
Sebastien Farnaud ◽  
...  
Keyword(s):  
Cell Lines ◽  
Transferrin Receptor ◽  
Iron Status ◽  
Hepatoma Cell ◽  
Human Hepatocytes ◽  
Mrna Levels ◽  
Hepcidin Expression ◽  
Hepatoma Cell Lines ◽  
Primary Human Hepatocytes ◽  
Transferrin Receptor 2

Hepcidin expression in vivo is regulated in proportion to iron status (i.e., increased by iron loading and decreased in iron deficiency). However, in vitro studies with hepatoma cell lines often show an inverse relationship between iron status and hepcidin expression. Here, we investigated possible molecular mechanisms responsible for the differences in iron sensing between hepatoma cell lines and human primary hepatocytes. RNA was collected from primary human hepatocytes, and HepG2 and HuH7 hepatoma cells were treated with either transferrin-bound and non-transferrin-bound iron. Expression of hepcidin, transferrin receptor 2, HFE, and hemojuvelin were quantified by real-time PCR. Hepcidin expression was increased in primary human hepatocytes following 24-h exposure to holoferric transferrin. In contrast, hepcidin mRNA levels in hepatoma cells were decreased by transferrin. Hepcidin expression was positively correlated with transferrin receptor 2 mRNA levels in primary human hepatocytes. Compared with primary hepatocytes, transferrin receptor 2 expression was significantly lower in hepatoma cell lines; furthermore, there was no correlation between transferrin receptor 2 and hepcidin mRNA levels in either HepG2 or HuH7 cells. Taken together our data suggest that transferrin receptor 2 is a likely candidate to explain the differences in iron sensing between hepatoma cell lines and primary human hepatocytes.

scholarly journals Distinctive HBV Replication Capacity and Susceptibility to Tenofovir Induced by a Polymerase Point Mutation in Hepatoma Cell Lines and Primary Human Hepatocytes

2021 ◽  
Vol 22 (4) ◽  
pp. 1606
Author(s):  
Ah Ram Lee ◽  
Ju-Yeon Cho ◽  
Jong Chul Kim ◽  
Mehrangiz Dezhbord ◽  
Soo Yeun Choo ◽  
...  
Keyword(s):  
Hepatitis B ◽  
Viral Replication ◽  
Cell Lines ◽  
Hepatoma Cell ◽  
Human Hepatocytes ◽  
Viral Fitness ◽  
High Rate ◽  
Replication Capacity ◽  
Hepatoma Cell Lines ◽  
Primary Human Hepatocytes

Tenofovir disoproxil fumarate (TDF) has been regarded as the most potent drug for treating patients with chronic hepatitis B (CHB). However recently, viral mutations associated with tenofovir have been reported. Here, we found a CHB patient with suboptimal response after more than 4 years of TDF treatment. Clonal analysis of hepatitis B virus (HBV) isolated from sequential sera of this patient identified the seven previously reported TDF-resistant mutations (CYELMVI). Interestingly, a threonine to alanine mutation at the 301 amino acid position of the reverse-transcriptase (RT) domain, (rtT301A), was commonly accompanied with CYELMVI at a high rate (72.7%). Since the rtT301A mutation has not been reported yet, we investigated the role of this naturally occurring mutation on the viral replication and susceptibility to tenofovir in various liver cells (hepatoma cells as well as primary human hepatocytes). A cell-based phenotypic assay revealed that the rtT301A mutation dramatically impaired the replication ability with meaningful reduction in sensitivity to tenofovir in hepatoma cell lines. However, attenuated viral replication by the rtT301A mutation was significantly restored in primary human hepatocytes (PHHs). Our findings suggest that the replication capability and drug sensitivity of HBV is different between hepatoma cell lines and PHHs. Therefore, our study emphasizes that validation studies should be performed not only in the liver cancer cell lines but also in the PHHs to understand the exact viral fitness under antiviral pressure in patients.

scholarly journals Correction: Permissivity of Primary Human Hepatocytes and Different Hepatoma Cell Lines to Cell Culture Adapted Hepatitis C Virus

PLoS ONE ◽  
2019 ◽  
Vol 14 (9) ◽  
pp. e0223022
Author(s):  
Francois Helle ◽  
Etienne Brochot ◽  
Carole Fournier ◽  
Véronique Descamps ◽  
Laure Izquierdo ◽  
...  
Keyword(s):  
Cell Culture ◽  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Cell Lines ◽  
Hepatoma Cell ◽  
Human Hepatocytes ◽  
Hepatoma Cell Lines ◽  
Primary Human Hepatocytes

scholarly journals Permissivity of Primary Human Hepatocytes and Different Hepatoma Cell Lines to Cell Culture Adapted Hepatitis C Virus

PLoS ONE ◽  
2013 ◽  
Vol 8 (8) ◽  
pp. e70809 ◽  
Author(s):  
Francois Helle ◽  
Etienne Brochot ◽  
Carole Fournier ◽  
Véronique Descamps ◽  
Laure Izquierdo ◽  
...  
Keyword(s):  
Cell Culture ◽  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Cell Lines ◽  
Hepatoma Cell ◽  
Human Hepatocytes ◽  
Hepatoma Cell Lines ◽  
Primary Human Hepatocytes

Cell-specific toxicity of a chimeric protein composed of interleukin-6 and Pseudomonas exotoxin (IL6-PE40) on tumor cells

1990 ◽  
Vol 10 (6) ◽  
pp. 2443-2447
Author(s):  
C B Siegall ◽  
R P Nordan ◽  
D J FitzGerald ◽  
I Pastan
Keyword(s):  
Cell Lines ◽  
Interleukin 6 ◽  
Hepatoma Cell ◽  
Chimeric Protein ◽  
Peptide Bond ◽  
Myeloma Cell ◽  
Mrna Levels ◽  
Pseudomonas Exotoxin ◽  
Chain Reactions ◽  
Hepatoma Cell Lines

IL6-PE40 is a chimeric toxin composed of human interleukin-6 (IL6) linked by a peptide bond to PE40, a form of Pseudomonas exotoxin (PE) devoid of its cell recognition domain. To identify cancer cell lines with high numbers of IL6 receptors and to assess the usefulness of IL6-PE40 as a possible anticancer agent, we evaluated the toxicity of IL6-PE40 on a variety of tumor cell lines and demonstrated that certain human myeloma and hepatoma cell lines were particularly sensitive. IL6 binding to selected hepatoma and myeloma cell lines were determined by using [125I]IL6. IL6 receptor mRNA levels were measured by polymerase chain reactions. When comparisons were made among different hepatoma cell lines, the sensitivity to IL6-PE40 correlated with the number of IL6 receptors. However, the hepatoma line PLC/PRF/5, which contains 2,300 IL6 receptors, was more sensitive to IL6-PE40 (amount of protein required to inhibit protein synthesis by 50% was 5 ng/ml) than both the myeloma cell lines U266 and H929 (for both cell lines, the 50% inhibitory dose was 8 ng/ml), which contain 15,500 and 16,500 IL6 receptors, respectively. RNA analysis confirmed that the sensitivity of these cells to IL6-PE40 and the amount of IL6 receptor RNA detected did not correlate. These data suggest that factors in addition to the number of IL6-binding sites contribute to the sensitivity of cells to IL6-PE40.

scholarly journals Chromatin structures of the rat tyrosine aminotransferase gene relate to the function of its cis-acting elements.

1990 ◽  
Vol 10 (7) ◽  
pp. 3334-3342 ◽  
Author(s):  
D Nitsch ◽  
A F Stewart ◽  
M Boshart ◽  
R Mestril ◽  
F Weih ◽  
...  
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Hepatoma Cell ◽  
Tyrosine Aminotransferase ◽  
Transient Transfection ◽  
Negative Regulator ◽  
Mrna Levels ◽  
Cell Type ◽  
Hepatoma Cell Lines ◽  
Cell Type Specific

The relationship between DNase I-hypersensitive sites (HSs) and transcriptional enhancers of the rat tyrosine aminotransferase (TAT) gene was examined by comparing HSs in and around the TAT gene with the activity of the corresponding DNA sequences in transient transfection assays. In this manner, we identified two HSs as liver-specific enhancers. Of three hepatoma cell lines examined, only one sustained TAT mRNA levels comparable to those of liver. In this cell line, both enhancers were strongly active, and strong hypersensitivity in chromatin over the enhancers was evident. The other two hepatoma cell lines had reduced levels of TAT mRNA and no or altered hypersensitivity over either the enhancers or the promoter. One of these lines carried a negative regulator of the TAT gene, the tissue specific extinguisher Tse-1. This cell line exhibited all HSs characteristic of the strongly active gene except at the promoter; however, one enhancer was inactive even though hypersensitive in chromatin. In a TAT-nonexpressing cell line, inactivity of both enhancers correlated with absence of the respective HSs. We conclude that although hypersensitivity in chromatin necessarily accompanies cell-type-specific enhancer activity, the occurrence of cell-type-specific HSs does not imply that the underlying sequences harbor enhancers active in transient transfection assays.

Inducible anti-sense RNA for angiotensinogen in stably transformed hepatoma cell lines

1990 ◽  
Vol 4 (2) ◽  
pp. 107-117 ◽  
Author(s):  
W. M. Clouston ◽  
C. J. Lloyd ◽  
R. I. Richards
Keyword(s):  
Cell Lines ◽  
Reductase Activity ◽  
Hepatoma Cell ◽  
Shuttle Vector ◽  
Mrna Level ◽  
Mrna Levels ◽  
Hepatoma Cell Lines ◽  
Metallothionein Promoter ◽  
Clonal Cell Lines ◽  
Fold Induction

ABSTRACT Angiotensinogen mRNA is found in many extrahepatic tissues, where it may participate in local angiotensin-generating systems. In this study we explore the feasibility of using anti-sense RNA to decrease angiotensinogen production in rat H4IIEC3 hepatoma cells. An amplifiable shuttle vector was modified to allow the production of high levels of stable anti-sense RNA from two regions of the mouse angiotensinogen gene under the control of the inducible sheep metallothionein promoter. Stably transformed, clonal cell lines expressing anti-sense RNA for angiotensinogen were isolated after selection with the aminoglycoside G418. Subsequently, the number of chromosomally integrated copies of the angiotensinogen anti-sense constructs was coamplified by methotrexate selection for dihydrofolate reductase activity carried on the shuttle vector. With a 20- to 30-fold induction of the anti-sense RNAs, the target angiotensinogen mRNA level was reduced to 25–30% of control values. The specificity of this effect was confirmed by showing no decrease in either β-tubulin or neomycin phosphotransferase mRNA levels. Using tissue-specific promoters, it should be possible to direct these effects to specific organs in transgenic mice. However, in agreement with results from other groups, our findings suggest that it will not be possible to eradicate completely the target gene product using the anti-sense RNA strategy.

Cytochrome P450 expression in human hepatocytes and hepatoma cell lines: molecular mechanisms that determine lower expression in cultured cells

Xenobiotica ◽  
2002 ◽  
Vol 32 (6) ◽  
pp. 505-520 ◽  
Author(s):  
C. Rodríguez-Antona ◽  
M. T. Donato ◽  
A. Boobis ◽  
R. J. Edwards ◽  
P. S. Watts ◽  
...  
Keyword(s):  
Cytochrome P450 ◽  
Cell Lines ◽  
Molecular Mechanisms ◽  
Cultured Cells ◽  
Hepatoma Cell ◽  
Human Hepatocytes ◽  
Hepatoma Cell Lines ◽  
P450 Expression ◽  
Lower Expression
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