Journal articles: 'HuH-7'

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Vecchi, Chiara, Giuliana Montosi, and Antonello Pietrangelo. "Huh-7: A human “hemochromatotic” cell line." Hepatology 51, no. 2 (October 27, 2009): 654–59. http://dx.doi.org/10.1002/hep.23410.

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Koutsoudakis, George, Eva Herrmann, Stephanie Kallis, Ralf Bartenschlager, and Thomas Pietschmann. "The Level of CD81 Cell Surface Expression Is a Key Determinant for Productive Entry of Hepatitis C Virus into Host Cells." Journal of Virology 81, no. 2 (November 1, 2006): 588–98. http://dx.doi.org/10.1128/jvi.01534-06.

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ABSTRACT Recently a cell culture model supporting the complete life cycle of the hepatitis C virus (HCV) was developed. Searching for host cell determinants involved in the HCV replication cycle, we evaluated the efficiency of virus propagation in different Huh-7-derived cell clones. We found that Huh-7.5 cells and Huh7-Lunet cells, two former replicon cell clones that had been generated by removal of an HCV replicon by inhibitor treatment, supported comparable levels of RNA replication and particle production, whereas virus spread was severely impaired in the latter cells. Analysis of cell surface expression of CD81 and scavenger receptor class B type I (SR-BI), two molecules previously implicated in HCV entry, revealed similar expression levels for SR-BI, while CD81 surface expression was much higher on Huh-7.5 cells than on Huh7-Lunet cells. Ectopic expression of CD81 in Huh7-Lunet cells conferred permissiveness for HCV infection to a level comparable to that for Huh-7.5 cells. Modulation of CD81 cell surface density in Huh-7.5 cells by RNA interference indicated that a certain amount of this molecule (∼7 × 104 molecules per cell) is required for productive infection with a low dose of HCV. Consistent with this, we show that susceptibility to HCV infection depends on a critical quantity of CD81 molecules. While infection is restricted in cells expressing very small amounts of CD81, susceptibility rapidly rises within a narrow range of CD81 levels, reaching a plateau where higher expression does not further increase the efficiency of infection. Together these data indicate that a high density of cell surface-exposed CD81 is a key determinant for productive HCV entry into host cells.

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Liu, Siyu, Minzi Yao, Xiaolong Li, Yumei Liu, and Cuiling Xie. "Effects of circFOXO3 on the Proliferation and Invasion of Liver Cancer Cells by Regulating PI3K/Akt Pathway." Contrast Media & Molecular Imaging 2022 (July 14, 2022): 1–7. http://dx.doi.org/10.1155/2022/2109908.

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Objective. Hepatocellular carcinoma is a malignant disease occurring in the liver and is one of the main causes of death in cancer patients. Tumor cells are the main components of tumors and have a strong proliferative capacity. They are easily transferred to other parts of the body and can produce harmful substances that destroy the normal organ structure and endanger human life and health. In this study, we investigate the effect of circFOXO3 on the proliferation and invasion of hepatocellular carcinoma cells and its possible mechanism. Methods. Human hepatocellular carcinoma cells BEL-7404, Hep G2, Hep 3B2.1–7, HuH-7, Li-7, and human normal hepatocytes HHL-5 were selected, and the expression level of circFOXO3 in the cell lines was determined by qRT-PCR. The cell line with low circFOXO3 expression level (HuH-7 cells) was used for follow-up experiments. HuH-7 liver cancer cells were divided into the control group (normal cultured), circFOXO3-NC group (transfected with circFOXO3 negative control), circFOXO3 mimic group (transfected with circFOXO3 mimic), PI3K activator group (20 μmol/L PI3K activator 740Y-P), and circFOXO3 mimic + PI3K activator group (transfected with circFOXO3 mimic + treated with PI3K activator 740Y-P). The qRT-PCR method was used to determine the expression level of circFOXO3 in HuH-7 liver cancer cells in each group, WB was used to detect the expression of apoptosis, invasion, and phosphoinositide 3-kinase/protein kinase B (PI3K/Akt) pathway related proteins in HuH-7 liver cancer cells in each group, the CCK-8 method was used to determine the viability of HuH-7 liver cancer cells in each group, flow cytometry was used to determine the apoptotic ability of HuH-7 liver cancer cells in each group, the transwell chamber experiment was used to determine the invasion ability of HuH-7 liver cancer cells in each group, and the scratch test was used to determine the migration ability of HuH-7 liver cancer cells in each group. Results. circFOXO3 showed low expression in liver cancer cells; compared with the control group, the circFOXO3 expression and apoptosis rate of HuH-7 liver cancer cells in the circFOXO3 mimic group were significantly increased ( P < 0.05 ) and the PI3K/Akt pathway-related protein expression, cell viability, invasion, and migration abilities were significantly reduced ( P < 0.05 ); the apoptosis rate of HuH-7 liver cancer cells in the PI3K activator group was significantly reduced ( P < 0.05 ) and the PI3K/Akt pathway related protein expression, cell viability, invasion and migration abilities were significantly increased ( P < 0.05 ). Compared with the circFOXO3 mimic group, the apoptosis rate of HuH-7 liver cancer cells in the circFOXO3 mimic + PI3K activator group was significantly reduced ( P < 0.05 ) and the PI3K/Akt pathway-related protein expression, cell viability, invasion and migration abilities were significantly increased ( P < 0.05 ). Conclusion. Highly expressed circFOXO3 can inhibit the proliferation and invasion of HuH-7 liver cancer cells, which may be achieved by inhibiting the PI3K/Akt pathway.

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Alsubaie, Saba M., Daoud Ali, Bader O. Almutairi, Rafa Almeer, and Saud Alarifi. "Evaluation of Cyto - and Genotoxic Influence of Lanthanum Dioxide Nanoparticles on Human Liver Cells." Dose-Response 20, no. 3 (July 2022): 155932582211284. http://dx.doi.org/10.1177/15593258221128428.

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Inorganic nanoparticles are representing an emerging paradigm in molecular imaging probe design. We have determined lanthanum oxide nanoparticles (La2O3 NPs)-induced toxicity on human livers cells for 48 hrs. Before exposure to La2O3 NPs, the size and shape of NPs were confirmed by transmission electron microscope. It was found at 32 ±1.6 nm with a sheet-like morphological structure. The viability of CHANG and HuH-7 cells was reduced as the concentration of La2O3 NPs increased. HuH-7 cells were more sensitive than CHANG cells to La2O3 NPs. We observed production of intracellular ROS in HuH-7 cells was more than CHANG cells and the LPO level was more in CHANG cells than in HuH-7 cells at 50 μg/ml of La2O3 NPs. Glutathione was decreased and catalase was increased at 50 μg/ml of La2O3 NPs. More apoptotic and necrotic cells were observed at 300 μg/ml in HuH-7 cells FACS. DNA damage was observed by the SGCE test and more DNA damage was found in CHANG cells than HuH-7 cells at 300 μg/ml La2O3 NPs over 48 hrs. Thus, study warrants the application of La2O3 NPs in daily life and provides vital information about the toxicity of La2O3 NPs.

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Schemmerer, Mathias, Monika Erl, and Jürgen J. Wenzel. "HuH-7-Lunet BLR Cells Propagate Rat Hepatitis E Virus (HEV) in a Cell Culture System Optimized for HEV." Viruses 14, no. 5 (May 23, 2022): 1116. http://dx.doi.org/10.3390/v14051116.

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The family Hepeviridae comprises the species Orthohepevirus A–D (HEV-A to -D). HEV-C genotype 1 (HEV-C1, rat HEV) is able to infect humans. This study investigated whether an optimized HEV-A cell culture system is able to propagate the cell culture-derived rat HEV, and if de novo isolation of the virus from rat liver is possible. We tested the liver carcinoma cell lines PLC/PRF/5, HuH-7, and HuH-7-Lunet BLR for their susceptibility to HEV-C1 strains. Cells were infected with the cell culture-derived HEV-C1 strain R63 and rat liver-derived strain R68. Cells were maintained in MEMM medium, which was refreshed every 3–4 days. The viral load of HEV-C1 was determined by RT-qPCR in the supernatant and expressed as genome copies per mL (c/mL). Rat HEV replication was most efficient in the newly introduced HuH-7-Lunet BLR cell line. Even if the rat HEV isolate had been pre-adapted to PLC/PRF/5 by multiple passages, replication in HuH-7-Lunet BLR was still at least equally effective. Only HuH-7-Lunet BLR cells were susceptible to the isolation of HEV-C1 from the liver homogenate. These results suggest HuH-7-Lunet BLR as the most permissive cell line for rat HEV. Our HEV-C1 cell culture system may be useful for basic research, the animal-free generation of large amounts of the virus as well as for the testing of antiviral compounds and drugs.

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Almasoud, Hadeel A., Daoud Ali, Khadijah N. Yaseen, Hanouf Almukhlafi, Norah S. Alothman, Bader Almutairi, Rafa Almeer, Nouf Alyami, Saad Alkahtani, and Saud Alarifi. "Dose-Dependent Variation in Anticancer Activity of Hexane and Chloroform Extracts of Field Horsetail Plant on Human Hepatocarcinoma Cells." BioMed Research International 2022 (June 25, 2022): 1–8. http://dx.doi.org/10.1155/2022/5778411.

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Horsetail fern plant is botanically known as Equisetum arvense L., and it is a good source of phenolic flavonoids, phenolic acids, and compounds. Anticancer properties of hexane and chloroform extracts of the horsetail fern plant and their mechanisms involved in the anticancer activity on human hepatocarcinoma (HuH-7) cells were examined. Cytotoxicity was evaluated by using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) and NRU (neutral red uptake) assays. Other parameters such as oxidative stress and apoptosis in pretreated hexane and chloroform extracts of the horsetail fern plant were examined in HuH-7 cells. The observation showed that hexane and chloroform extract of the horsetail fern plant exhibited cytotoxicity against HuH-7 cells. The value of IC50-24 h of hexane and chloroform extract of the horsetail fern plant was determined as 199.0 μg/ml and 161.90 0 μg/ml for HuH-7 cells, respectively, and on the basis of IC50 value, three acute concentrations, viz., 75% of IC50, 50% of IC50, and 25% of IC50, were determined for further study. The lower dose of extracts hexane and chloroform extract of the horsetail fern plant did not show significant toxicity. Higher concentrations of extract induced significant antioxidant effects as well as apoptosis effects. However, exposure to hexane and chloroform extract of the horsetail fern plant upregulated the expression of Bax and p53 in HuH-7 cells. These data suggest that hexane and chloroform extract of the horsetail fern plant plays a significant role in the induction of toxicity via the regulation of oxidative stress in HuH-7 cells. This work may be useful for cancer chemotherapy.

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Katubi, KM, FM Alzahrani, D. Ali, and S. Alarifi. "Dose- and duration-dependent cytotoxicity and genotoxicity in human hepato carcinoma cells due to CdTe QDs exposure." Human & Experimental Toxicology 38, no. 8 (April 17, 2019): 914–26. http://dx.doi.org/10.1177/0960327119843578.

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Nanotechnology has achieved more commercial attention over recent years, and its application has increased concerns about its discharge in the environment. In this study, we have chosen human hepatic carcinoma (HuH-7) cells because liver tissue has played an important role in human metabolism. Therefore, the objective of this study was to determine DNA damaging and apoptotic potential of cadmium telluride quantum dots (CdTe QDs; average particle size (APS) 10 nm, 1–25 µg/ml) on HuH-7 cells and the basic molecular mechanism of its cellular toxicity. Cytotoxicity of different concentrations of CdTe QDs on HuH-7 cells was determined by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) and lactate dehydrogenase (LDH) tests. Moreover, reactive oxygen species (ROS) generation, mitochondrial membrane potential, DNA damage, and Hoechst 33342 fluorescent staining morphological analysis of necrotic/apoptotic cells were detected; cellular impairment in mitochondria and DNA was confirmed by JC-1 and comet assay, respectively. A dose- and time-dependent cytotoxicity effect of CdTe QDs exposure was observed HuH-7 cells; the significant ( p < 0.05) cytotoxicity was found at 25 μg/ml of CdTe QDs exposure. The percentage of cytotoxicity of CdTe QDs (25 μg/ml) in HuH-7 cells reached 62% in 48 h. CdTe QDs elicited intracellular ROS generation and mitochondrial depolarization, and DNA integrity cells collectively advocated the apoptotic cell death at higher concentration. DNA damage was observed in cells due to CdTe QDs exposure, which was mediated by oxidative stress. This study exploring the effects of CdTe QDs in HuH-7 cells has provided valuable insights into the mechanism of toxicity induced by CdTe QDs.

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Hada, Toshikazu, Tetsuya Yamamoto, Hiroyasu Imanishi, Sumio Takahashi, Yoshiki Amuro, Kazuya Higashino, and Jiro Sato. "Novel Cholinesterase Expression in the HuH-7 Cell Line." Tumor Biology 8, no. 1 (1987): 3–8. http://dx.doi.org/10.1159/000217485.

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Zhao, Dan, Meigeng Hu, Guoxu Ma, and Xudong Xu. "Five New Terpenes with Cytotoxic Activity from Pestalotiopsis sp." Molecules 26, no. 23 (November 29, 2021): 7229. http://dx.doi.org/10.3390/molecules26237229.

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Five new compounds called Pestalotis A–E (1–5), comprising three monoterpene-lactone compounds (1–3), one tetrahydrobenzofuran derivative (4), and one sesquiterpene (5), were isolated from the EtOAc extract of Pestalotiopsis sp. The structures of the new compounds were elucidated by analysis of their NMR, HRMS, and ECD spectra, and the absolute configurations were established through the comparison of experimental and calculated ECD spectra. All compounds were tested for antitumor activity against SW-480, LoVo, HuH-7, and MCF-7. The results showed that compounds 2 and 4 exhibited potent antitumor activity against SW-480, LoVo, and HuH-7 cell lines. Furthermore, compound 4 was assessed against HuH-7, and the results indicated that the rate of apoptosis was dose-dependent.

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Franco, Evelyn J., Camilly P. Pires de Mello, and Ashley N. Brown. "Antiviral Evaluation of UV-4B and Interferon-Alpha Combination Regimens against Dengue Virus." Viruses 13, no. 5 (April 27, 2021): 771. http://dx.doi.org/10.3390/v13050771.

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Dengue virus (DENV) is a flavivirus associated with clinical manifestations ranging in severity from self-limiting dengue fever, to the potentially life threatening condition, severe dengue. There are currently no approved antiviral therapies for the treatment of DENV. Here, we evaluated the antiviral potential of four broad-spectrum antivirals, UV-4B, interferon-alpha (IFN), sofosbuvir (SOF), and favipiravir (FAV) against DENV serotype 2 as mono- and combination therapy in cell lines that are physiologically relevant to human infection. Cell lines derived from human liver (HUH-7), neurons (SK-N-MC), and skin (HFF-1) were infected with DENV and treated with UV-4B, IFN, SOF, or FAV. Viral supernatant was sampled daily and infectious viral burden was quantified by plaque assay on Vero cells. Drug effect on cell proliferation in uninfected and infected cells was also assessed. UV-4B inhibited DENV in HUH-7, SK-N-MC, and HFF-1 cells yielding EC50 values of 23.75, 49.44, and 37.38 µM, respectively. Clinically achievable IFN concentrations substantially reduced viral burden in HUH-7 (EC50 = 102.7 IU/mL), SK-N-MC (EC50 = 86.59 IU/mL), and HFF-1 (EC50 = 163.1 IU/mL) cells. SOF potently inhibited DENV in HUH-7 cells but failed to produce the same effect in SK-N-MC and HFF-1 cells. Finally, FAV provided minimal suppression in HUH-7 and SK-N-MC cells, but was ineffective in HFF-1 cells. The two most potent anti-DENV agents, UV-4B and IFN, were also assessed in combination. UV-4B + IFN treatment enhanced antiviral activity in HUH-7, SK-N-MC, and HFF-1 cells relative to monotherapy. Our results demonstrate the antiviral potential of UV-4B and IFN against DENV in multiple physiologically relevant cell types.

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Logue, James, Walter Vargas Licona, Timothy Cooper, Becky Reeder, Russel Byrum, Jing Qin, Nicole Deiuliis Murphy, et al. "Ebola Virus Isolation Using Huh-7 Cells has Methodological Advantages and Similar Sensitivity to Isolation Using Other Cell Types and Suckling BALB/c Laboratory Mice." Viruses 11, no. 2 (February 16, 2019): 161. http://dx.doi.org/10.3390/v11020161.

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Following the largest Ebola virus disease outbreak from 2013 to 2016, viral RNA has been detected in survivors from semen and breast milk long after disease recovery. However, as there have been few cases of sexual transmission, it is unclear whether every RNA positive fluid sample contains infectious virus. Virus isolation, typically using cell culture or animal models, can serve as a tool to determine the infectivity of patient samples. However, the sensitivity of these methods has not been assessed for the Ebola virus isolate, Makona. Described here is an efficiency comparison of Ebola virus Makona isolation using Vero E6, Huh-7, monocyte-derived macrophage cells, and suckling laboratory mice. Isolation sensitivity was similar in all methods tested. Laboratory mice and Huh-7 cells were less affected by toxicity from breast milk than Vero E6 and MDM cells. However, the advantages associated with isolation in Huh-7 cells over laboratory mice, including cost effectiveness, sample volume preservation, and a reduction in animal use, make Huh-7 cells the preferred substrate tested for Ebola virus Makona isolation.

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Ali, Samir, Charles Pellerin, Daniel Lamarre, and George Kukolj. "Hepatitis C Virus Subgenomic Replicons in the Human Embryonic Kidney 293 Cell Line." Journal of Virology 78, no. 1 (January 1, 2004): 491–501. http://dx.doi.org/10.1128/jvi.78.1.491-501.2004.

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ABSTRACT Hepatitis C virus (HCV) infects liver cells and its replication in other cells is incompletely defined. Human hepatoma Huh-7 cells harboring subgenomic HCV replicons were used in somatic cell fusion experiments with human embryonic kidney 293 cells as a means of examining the permissiveness of 293 cells for HCV subgenomic RNA replication. 293 cells were generally not permissive for replication of Huh-7 cell-adapted replicons. However, upon coculturing of the two cell lines, we selected rare replicon-containing cells, termed 293Rep cells, that resembled parental 293 cells. Direct metabolic labeling of cells with 33P in the presence of actinomycin D and Northern blotting to detect the negative strand of the replicon demonstrated functional RNA replicons in 293Rep cells. Furthermore, Western blots revealed that 293Rep cells expressed the HCV nonstructural proteins as well as markers of the naïve 293 cells but not Huh-7 cells. Propidium iodide staining and fluorescence-activated cell sorting analysis of 293Rep cells revealed that clone 293Rep17 closely resembled naïve 293 cells. Transfection of total RNA from 293Rep17 into naïve 293 cells produced replicon-containing 293 cell lines with characteristics distinct from those of Huh-7-derived replicon cell lines. Relative to Huh-7 replicons, the 293 cell replicons were less sensitive to inhibition by alpha interferon and substantially more sensitive to inhibition by poly(I)-poly(C) double-stranded RNA. This study established HCV subgenomic replicons in nonhepatic 293 cells and demonstrated their utility in expanding the study of cellular HCV RNA replication.

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Li, Xiao-Hong, Kristine C. Y. McGrath, Van H. Tran, Yi-Ming Li, Sravan Mandadi, Colin C. Duke, Alison K. Heather, and Basil D. Roufogalis. "Identification of a Calcium Signalling Pathway ofS-[6]-Gingerol in HuH-7 Cells." Evidence-Based Complementary and Alternative Medicine 2013 (2013): 1–7. http://dx.doi.org/10.1155/2013/951758.

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Calcium signals in hepatocytes control cell growth, proliferation, and death. Members of the transient receptor potential (TRP) cation channel superfamily are candidate calcium influx channels. NFκB activation strictly depends on calcium influx and often induces antiapoptotic genes favouring cell survival. Previously, we reported thatS-[6]-gingerol is an efficacious agonist of the transient receptor potential cation channel subfamily V member 1 (TRPV1) in neurones. In this study, we tested the effect ofS-[6]-gingerol on HuH-7 cells using the Fluo-4 calcium assay, RT-qPCR, transient cell transfection, and luciferase measurements. We found thatS-[6]-gingerol induced a transient rise in[Ca2+]iin HuH-7 cells. The increase in[Ca2+]iinduced byS-[6]-gingerol was abolished by preincubation with EGTA and was also inhibited by the TRPV1 channel antagonist capsazepine. Expression of TRPV1 in HuH-7 cells was confirmed by mRNA analysis as well as a test for increase of[Ca2+]iby TRPV1 agonist capsaicin and its inhibition by capsazepine. We found thatS-[6]-gingerol induced rapid NFκB activation through TRPV1 in HuH-7 cells. Furthermore,S-[6]-gingerol-induced NFκB activation was dependent on the calcium gradient and TRPV1. The rapid NFκB activation byS-[6]-gingerol was associated with an increase in mRNA levels of NFκB-target genes: cIAP-2, XIAP, and Bcl-2 that encode antiapoptotic proteins.

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Yamashita, Yo-Ichi, Mitsuo Shimada, Norifumi Harimoto, Shinji Tanaka, Ken Shirabe, Hiroyuki Ijima, Kohji Nakazawa, Junji Fukuda, Kazumori Funatsu, and Yoshihiko Maehara. "cDNA Microarray Analysis in Hepatocyte Differentiation in Huh 7 Cells." Cell Transplantation 13, no. 7-8 (October 2004): 793–800. http://dx.doi.org/10.3727/000000004783983396.

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Yang, Dongqin, Takahiro Yaguchi, Tetsu Nagata, Akinobu Gotoh, Sinisa Dovat, Chunhua Song, and Tomoyuki Nishizaki. "AMID Mediates Adenosine-Induced Caspase-Independent HuH-7 Cell Apoptosis." Cellular Physiology and Biochemistry 27, no. 1 (2011): 37–44. http://dx.doi.org/10.1159/000325203.

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Mansoor, Tayyab A., Rita M. Ramalho, Xuan Luo, Cátia Ramalhete, Cecília M. P. Rodrigues, and Maria-José U. Ferreira. "Isoflavones as Apoptosis Inducers in Human Hepatoma HuH-7 Cells." Phytotherapy Research 25, no. 12 (April 14, 2011): 1819–24. http://dx.doi.org/10.1002/ptr.3498.

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Lee, Gigi, and Micheline Piquette-Miller. "Influence of IL-6 on MDR and MRP-mediated multidrug resistance in human hepatoma cells." Canadian Journal of Physiology and Pharmacology 79, no. 10 (October 1, 2001): 876–84. http://dx.doi.org/10.1139/y01-071.

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The objective of this study was to examine effects of interleukin-6 (IL-6) on the expression and activity of the drug resistance transporters (MDR1 and MRP) in human hepatoma cell lines. Expression and activity of MDR1 and MRP transporters were examined in IL-6-treated and control HuH 7 and HepG2 cells using semi-quantitative RT-PCR analysis and by rhodamine 123 and 5-carboxyfluorescin efflux assays. Results from RT-PCR demonstrated expression of MRP3, MRP6, and MDR1 in HuH 7 cells and expression of MRP1, MRP2, MRP3, MRP6, and MDR1 in HepG2 cells. Compared with controls, treatment of HuH 7 cells with IL-6 (10 ng/mL, 24 h) resulted in a 1.8-fold increase in MRP-mediated efflux of 5-CF with a corresponding 1.5-fold induction of MRP3 mRNA levels (p < 0.05). Similarly, in HepG2 cells, a 2-fold increase in MRP functional activity and a 1.8-fold induction of MRP1 mRNA levels were seen in the IL-6 treated cells (p < 0.05). Treatment of cells with IL-6 was also found to cause significant reductions in the expression and activity of MDR1 in HuH 7 cells, but not in HepG2 cells. Our data suggest that IL-6 induces MRP expression and activity in human hepatoma cell lines. Suppressive effects of IL-6 on MDR1 expression and activity were also observed in HuH 7 cells. This underscores the importance of examining the regulation of multiple drug resistance proteins as these proteins may have opposing regulatory mechanisms in malignant cells.Key words: P-Glycoprotein, multidrug resistance proteins, hepatocarcinogenesis, cytokines, inflammation.

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Takeda, Midori, Masanori Ikeda, Yasuo Ariumi, Takaji Wakita, and Nobuyuki Kato. "Development of hepatitis C virus production reporter-assay systems using two different hepatoma cell lines." Journal of General Virology 93, no. 7 (July 1, 2012): 1422–31. http://dx.doi.org/10.1099/vir.0.040725-0.

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A hepatitis C virus (HCV) infection system was developed previously using the HCV JFH-1 strain (genotype 2a) and HuH-7 cells, and this cell culture is so far the only robust production system for HCV. In patients with chronic hepatitis C, the virological effects of pegylated interferon and ribavirin therapy differ depending on the HCV strain and the genetic background of the host. Recently, we reported the hepatoma-derived Li23 cell line, in which the JFH-1 life cycle is reproduced at a level almost equal to that in HuH-7-derived RSc cells. To monitor the HCV life cycle more easily, we here developed JFH-1 reporter-assay systems using both HuH-7- and Li23-derived cell lines. To identify any genetic mutations by long-term cell culture, HCV RNAs in HuH-7 cells were amplified 130 days after infection and subjected to sequence analysis to find adaptive mutation(s) for robust virus replication. We identified two mutations, H2505Q and V2995L, in the NS5B region. V2995L but not H2505Q enhanced JFH-1 RNA replication. However, we found that H2505Q but not V2995L enhanced HCV RNA replication of strain O (genotype 1b). We also selected highly permissive D7 cells by serial subcloning of Li23 cells. The expression levels of claudin-1 and Niemann–Pick C1-like 1 in D7 cells are higher than those in parental Li23 cells. In this study, we developed HCV JFH-1 reporter-assay systems using two distinct hepatoma cell lines, HuH-7 and Li23. The mutations in NS5B resulted in different effects on strains O and JFH-1 HCV RNA replication.

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Lin, Chuan-Yi, May-Hua Liao, Chi-Yu Yang, Chao-Kai Chang, Shih-Mei Hsu, Chi-Long Juang, and Hsiao-Chuan Wen. "Anti-Metastatic Activity of Tagitinin C from Tithonia diversifolia in a Xenograft Mouse Model of Hepatocellular Carcinoma." Livers 2, no. 4 (December 1, 2022): 400–411. http://dx.doi.org/10.3390/livers2040030.

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Sesquiterpenoid tagitinin C, present in Tithonia diversifolia leaves, has been known to have anti-hepatoma properties. Therefore, we investigated the anti-metastatic potential of tagitinin C in xenograft models of hepatocellular carcinoma (HCC). We isolated tagitinin C from a methanolic extract of the leaves of T. diversifolia. HepG-2 and Huh 7 hepatoma cells were treated with tagitinin C, and cell viability, migration, and matrix metalloproteinase (MPP) activity were assessed using the 3-(4,5-dimethylthiozol-2-yl)-2,5-diphenyltetrazolium bromide assay, scratch migration assay, and MMP activity assay, respectively. We used magnetic resonance spectroscopy to determine the tumorigenicity of xenografts inoculated with Hep-G2 and Huh 7 cells. Tagitinin C was cytotoxic against Hep-G2 and Huh 7 cells, with IC50 values of 2.0 ± 0.1 µg/mL and 1.2 ± 0.1 µg/mL, respectively, and it showed an anti-metastatic effect in vitro. Additionally, MRS assays revealed that tagitinin C (15 g/mouse/day) reduced the tumorigenicity of Hep-G2 and Huh 7 cell xenografts. Tagitinin C demonstrated significant antitumor and anti-metastatic activity in the two human hepatoma cell lines. Tagitinin C might be used as an alternative or auxiliary therapy for the treatment of HCC, and its effect should be further investigated in clinical settings.

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Nakabayashi, Hidekazu, and Kazuhisa Taketa. "HuH-7 cell line established from a highly differentiated human hepatocellular carcinoma." Okayama Igakkai Zasshi (Journal of Okayama Medical Association) 124, no. 3 (2012): 231–38. http://dx.doi.org/10.4044/joma.124.231.

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Müller, Stefanie, Robert Geffers, and Stephan Günther. "Analysis of gene expression in Lassa virus-infected HuH-7 cells." Journal of General Virology 88, no. 5 (May 1, 2007): 1568–75. http://dx.doi.org/10.1099/vir.0.82529-0.

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The pathogenesis of Lassa fever is poorly understood. As the liver is a major target organ of Lassa virus, gene expression in Lassa virus-infected HuH-7 cells, a differentiated human hepatoma cell line, was studied. Cellular mRNA levels were measured at the late phase of acute infection, when virtually all cells expressed large amounts of nucleoprotein, and virus RNA concentration had reached >108 copies (ml supernatant)−1. Two types of transcription array were used: cDNA-based macroarrays with a set of 3500 genes (Atlas Human 1.2 arrays; Clontech) and oligonucleotide-based microarrays covering 18 400 transcripts (Human Genome U133A array; Affymetrix). Data analysis was based on statistical frameworks controlling the false-discovery rate. Atlas array data were considered relevant if they could be verified by U133A array or real-time RT-PCR. According to these criteria, there was no evidence for true changes in gene expression. Considering the precision of the U133A array and the number of replicates tested, potential expression changes due to Lassa virus infection are probably smaller than twofold. To substantiate the array data, beta interferon (IFN-β) gene expression was studied longitudinally in Lassa virus-infected HuH-7 and FRhK-4 cells by using real-time RT-PCR. IFN-β mRNA levels increased only twofold upon Lassa virus infection, although there was no evidence that the virus inhibited poly(I : C)-induced IFN-β gene expression. In conclusion, Lassa virus interferes only minimally with gene expression in HuH-7 cells and poorly induces IFN-β gene transcription.

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Kolke, Chisato, Yumiko Hayakawa, Kenji Niiya, Nobuo Sakuragawa, and Hideo Sasaki. "The Production of Heparin Cofactor li Is Not Regulated by Inflammatory Cytokines in Human Hepatoma Cells: Comparison with Plasminogen Activator Inhibitor Type-1." Thrombosis and Haemostasis 75, no. 02 (1996): 298–302. http://dx.doi.org/10.1055/s-0038-1650264.

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SummaryUsing the Northern blot technique, we screened 6 human hepatoma cell lines to investigate the regulation mechanism of heparin cofactor II (HCII) biosynthesis. We found that HuH-7 and Hep G2 cells constitutively expressed the HC II gene. In conditioned medium, HuH-7 cells constantly produced HC II that was functionally active and formed a complex with thrombin in the presence of dermatan sulfate. HC II is thought be an acute phase reactant, and, therefore, we examined the effects of the major inflammatory cytokines, IL-6, IL-1β, and TNF-α, on the regulation of HC II production in HuH-7 and Hep G2 cells. In HuH-7 cells, the antigen and mRNA levels of plasminogen activator inhibitor type-1 (PAI-1), an acute phase protein produced by hepa-tocytes, were increased in response to stimulation with either IL-6 or IL-1 (3 or both, but HC II antigen and mRNA levels were not changed by the same stimulation. Even when Hep G2 cells were treated with a combination of three cytokines, IL-6, IL-1β, and TNF-α, HC II antigen and mRNA levels were not changed; however, PAI-1 antigen and mRNA levels were clearly increased. These results suggest that the production of HC II in hepatoma cells is not regulated by the major inflammatory mediators, IL-6, IL-iβ, and TNF-α.

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Bin-Jumah, May N., Monera Al-Abdan, Gadah Al-Basher, and Saud Alarifi. "Molecular Mechanism of Cytotoxicity, Genotoxicity, and Anticancer Potential of Green Gold Nanoparticles on Human Liver Normal and Cancerous Cells." Dose-Response 18, no. 2 (April 1, 2020): 155932582091215. http://dx.doi.org/10.1177/1559325820912154.

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Nanomaterials are extensively applied in various fields such as industry, medicine, and food and drugs due to their unique properties. In this study, gold nanoparticles were biosynthesized using leaf extract of Azadirachta indica and chloroauric acid salt. We have determined the cytotoxicity, genotoxicity, and apoptotic effect of green gold nanoparticles (gGNPs) on human normal (CHANG) and liver cancer (HuH-7) cells. Before exposure to cells, physiochemical characteristic of gGNPs was characterized using a transmission electron microscope and dynamic light scattering. Cytotoxicity of gGNPs was found dose-dependent, as it was confirmed using 2 methods, namely, 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide and neutral red uptake. The gGNPs provoked intracellular reactive oxygen species (ROS), lipid peroxide, and reduced total glutathione and mitochondrial membrane potential in CHANG and HuH-7 cells in a dose-dependent manner. We have observed that N-acetyl-l-cysteine inhibits the generation of ROS in both cells after exposure to gGNPs. DNA damaging effects of gGNPs were determined by comet assay, and the maximum DNA damage was observed at 700 µg/mL gGNPs for 24 hours. It was observed that HuH-7 cells are slightly more sensitive to gGNPs exposure than CHANG cells. In conclusion, cytotoxicity and apoptosis in CHANG and HuH-7 cells due to gGNPs were mediated through oxidative stress.

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Lee, Kwong-Chiu, Yao-Li Chen, Ping-Yi Lin, and Wan-Ling Chuang. "Ursolic Acid-Induced Apoptosis via Regulation of the PI3K/Akt and MAPK Signaling Pathways in Huh-7 Cells." Molecules 23, no. 8 (August 13, 2018): 2016. http://dx.doi.org/10.3390/molecules23082016.

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Ursolic acid (UA), is a kind of triterpene acid that exhibits wide biological properties. In this article, the effects of UA on apoptosis and the proliferation of human hepatoma Huh-7 cells were reported. The MTT results showed that cell viability of Huh-7 was reduced in a concentration and time-dependent effect. In addition, DAPI staining was used to detected condensation of chromatin in nucleus. Apoptotic cell population was examined using Annexin V/PI staining. The results showed that exposure to UA affected extrinsic and intrinsic pathways through, reduced expression of Bcl-2, Mcl-1, and TCTP; increased levels of the apoptotic proteins TNF-α, Fas, FADD, and Bax; and activation of cleaved caspase-3 and PARP. UA also inhibited the p-Akt and p38 MAPK signaling transduction pathways, and increased activity in the p-ERK signaling pathway. Taken together, UA inhibited the cell growth of Huh-7 cells and affected apoptosis, via regulated cellular signaling transduction.

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Enjolras, Nathalie, Yesim Dargaud, Jean-Luc Plantier, Heli Simpson, Florine Guillaume, and Claude Negrier. "Hepatoma Cell Line HuH-7 : a Promising New Cellular System for Recombinant FIX Production." Blood 114, no. 22 (November 20, 2009): 4202. http://dx.doi.org/10.1182/blood.v114.22.4202.4202.

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Abstract Abstract 4202 Hemophilia B is characterized by a deficiency of coagulation factor IX (FIX) which is synthesized by the liver. CHO cells commonly used for the production of recombinant coagulation factors have a limited capacity for post-translational modifications leading to incomplete γ-carboxylation, decreased phosphorylation and sulfation of the recombinant coagulation proteins. In addition, studies in animal models and data from clinical studies in hemophilic patients, have shown that recombinant FIX has a lower recovery than plasma-derived preparations. In the present study, the human hepatoma cell line HuH-7 was assessed for the production of a recombinant FIX molecule with post-translational modifications similar to those of the plasma-derived FIX. Methods Human hepatoma cell line HuH-7 was transfected with native FIX cDNA directed by CMV promoter, and cellular clones stably expressing FIX were obtained. Benefix®, Mononine® and a protein extract from human liver were used as controls. Cellular expression was analyzed by northern blot, and FIX production was quantified by ELISA. Procoagulant activities were measured by one-stage clotting and thrombin generation (TGT) assays. Secreted and intracellular FIX were visualized by western blot using an antibody directed against FIX. Intracellular trafficking of FIX was further analyzed using inhibitors of proteasome, lysosome and Golgi pathways. Glycosylation profile was determined after successive digestions of lysates by neuraminidase and N-Glycosydase-F. Results Non transfected HuH-7 cells did not express FIX and northern blot did not show any signal corresponding to FIX mRNA. Transiently transfected HuH-7 cells studies revealed the ability to secrete 0.1μg/ml/24h of FIX. The specific activity, measured by a one-stage clotting assay, reached 200% in comparison with Benefix® or rFIX produced by CHO cells. TGT confirmed the high procoagulant activity showing a 2.5 fold increased thrombin generating capacity in comparison with FIX produced by CHO cells, in the same experimental conditions. The secreted molecule appeared as a single band on SDS-PAGE, and presented the same mobility than Benefix® and Mononine®. Following enzymatic digestion, the migration profile suggested that glycosylation and sialylation were similar to Benefix®. Western blot showed that FIX from cell lysate was retrieved as three bands. The upper band, presenting a lower signal than the two others, had the same molecular weight than the secreted form and the native human FIX extracted from a liver biopsy. The two other bands at different intensities migrated slightly faster than the secreted protein. Brefeldin A significantly blocked intracellular traffic of FIX from ER to the Golgi complex where the protein corresponding to the western blot upper band was dramatically accumulated. No intracellular accumulation of the non-secreted forms was observed, and lysosomal pathway was slightly involved in the degradation of both secreted and non-secreted forms of FIX. Conclusion These data demonstrate that HuH-7 cell line is able to produce in serum free medium a biologically active recombinant FIX with a higher specific activity. The protein is correctly synthesized, follows the classical pathway before secretion and therefore is not retained in the cells. N-linked oligosaccharide sialylation is also correctly performed. This preliminary data suggest that HuH7 cells may represent an effective cellular system for rFIX production. Further biochemical analysis of the post-translational modifications and pharmacokinetic properties are required to better characterize the rFIX molecule secreted by HuH7 cells. Disclosures: No relevant conflicts of interest to declare.

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Fan, Guangsheng, Xiaoming Ma, Betsy T. Kren, and Clifford J. Steer. "Unbound E2F modulates TGF-β1-induced apoptosis in HuH-7 cells." Journal of Cell Science 115, no. 15 (August 1, 2002): 3181–91. http://dx.doi.org/10.1242/jcs.115.15.3181.

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E2F is an important target of the retinoblastoma protein (pRb) and plays a critical role in G1/S progression through the cell cycle. TGF-β1 arrests HuH-7 cells in G1 by suppressing phosphorylation of pRb and induces apoptosis by inhibiting its expression. In this study, we examined the downstream effects of TGF-β1-induced apoptosis and the potential roles for pRb and E2F. The results indicated that greater than 90% of the TGF-β1-induced preapoptotic cells were arrested in G1 phase of the cell cycle. This was associated with a significant increase in both E2F-DNA-binding activity and transcription of E2F-responsive reporter constructs. In contrast, no significant changes were observed in E2F mRNA and protein levels, and the overexpression of pRb partially inhibited E2F activation. Gel-shift assays identified more than four E2F complexes from preapoptotic and synchronized G1 HuH-7 cells,each exhibiting different patterns of E2F-associated proteins. The increased E2F activity did not affect the association patterns with pRb, p107 and p130,but altered the formation of an E2F—DP-1 complex. In contrast,E2F—DP-2 exhibited little change in the preapoptotic cells. Moreover,TGF-β1 induced apoptosis at G1 and inhibited entry into S phase irrespective of the increased E2F activity. The release of preapoptotic cells from TGF-β1 resulted in rapid S phase entry and subsequent apoptosis in 33% of cells over a 72 hour period. In conclusion, the results demonstrate that TGF-β1-induced apoptosis in HuH-7 cells is associated with a marked increase in activity of transcription factor E2F that is partially inhibited by overexpression of pRb. Preapoptotic changes are, in part, reversible upon removal of TGF-β1 and the majority of cells re-enter the normal cell cycle. Finally, TGF-β1-induced apoptosis with the associated increase in E2F activity can occur in both the G1and S phases of the cell cycle.

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Jouan, Elodie, Marc Le Vée, Claire Denizot, Yannick Parmentier, and Olivier Fardel. "Drug Transporter Expression and Activity in Human Hepatoma HuH-7 Cells." Pharmaceutics 9, no. 4 (December 28, 2016): 3. http://dx.doi.org/10.3390/pharmaceutics9010003.

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Wang, Chunjing, Chengcheng Li, and Rui Hao. "miR-559 Inhibits Proliferation, Autophagy, and Angiogenesis of Hepatocellular Carcinoma Cells by Targeting PARD3." Mediators of Inflammation 2022 (September 5, 2022): 1–9. http://dx.doi.org/10.1155/2022/3121492.

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Hepatocellular carcinoma (HCC) is one of the most common cancers in the world and has a high mortality rate. Although prevention and treatment of HCC has improved, it still faces poor prognosis and high mortality. miRNAs play a critical role in the tumorigenesis of HCC, but the underlying mechanism has not been well investigated. Here, the functions and interaction between miR-559 and PARD3 were investigated in HCC cells. Increased PARD3 and decreased miR-559 expression were observed in HCC cells compared with those in normal liver cells, especially in Huh-7 cells. Studies further demonstrated that PARD3 silencing or miR-559 overexpression impaired the proliferation, autophagy, and angiogenesis in Huh-7 cells. Mechanistically, PARD3 represents a target of miR-559. Furthermore, investigations revealed that miR-559 inhibition induced the expression of PARD3, thereby enhancing cell proliferation, autophagy, and angiogenesis in Huh-7 cells. These results reveal the interaction between miR-559 and PARD3 in HCC cells and provide new insights into their potential targets as therapeutic treatment against HCC.

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Wang, Ya-Jing, Nan Ma, Yong-Fu Lu, Si-Yang Dai, Xue Song, Chang Li, Yi Sun, and Yue-Hu Pei. "Structure Elucidation and Anti-Tumor Activities of Trichothecenes from Endophytic Fungus Fusariumsporotrichioides." Biomolecules 12, no. 6 (June 2, 2022): 778. http://dx.doi.org/10.3390/biom12060778.

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The secondary metabolites of Fusarium sporotrichioides, an endophytic fungus with anti-tumor activity isolated from Rauvolfia yunnanensis Tsiang, were investigated. Five trichothecenes, including one previously undescribed metabolite, were isolated and identified. Their structures were elucidated by means of extensive spectroscopic methods; the absolute configuration of compound 1 was determined by the ECD method. Surprisingly, 8-n-butyrylneosolaniol (3) exhibited stronger anti-tumor activity than T-2 toxin against Huh-7 cell line, with an IC50 value of 265.9 nM. 8-n-butyrylneosolaniol (3) promoted apoptosis induction in Huh-7 cells. Moreover, cell cycle analysis showed that cell cycle arrest caused by 8-n-butyrylneosolaniol (3) at the G2/M phase resulted in cell proliferation inhibition and pro-apoptotic activity. Further studies showed a significant decrease in mitochondrial membrane permeabilization and a significant increase in ROS generation, which led to the activation of caspase cascades and subsequent cleavage of PARP fragments. In conclusion, 8-n-butyrylneosolaniol (3) induced cell apoptosis in Huh-7 cells via the mitochondria-mediated apoptotic signaling pathway, which could be a leading compound for anti-tumor agents.

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Hernandez-Santiago, Brenda I., Thierry Beltran, Lieven Stuyver, Chung K. Chu, and Raymond F. Schinazi. "Metabolism of the Anti-Hepatitis C Virus Nucleoside β-d-N4-Hydroxycytidine in Different Liver Cells." Antimicrobial Agents and Chemotherapy 48, no. 12 (December 2004): 4636–42. http://dx.doi.org/10.1128/aac.48.12.4636-4642.2004.

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ABSTRACT β-d-N 4-Hydroxycytidine (NHC) was found to have selective anti-hepatitis C virus (HCV) activity in the HCV replicon system (clone A). The intracellular metabolism of tritiated NHC was investigated in the HCV replicon system, Huh-7 cells, HepG2 cells, and primary human hepatocytes. Incubation of cells with 10 μM radiolabeled NHC demonstrated extensive and rapid phosphorylation in all liver cells. Besides the 5′-mono, -di-, and -triphosphate metabolites of NHC, other metabolites were characterized. These included cytidine and uridine mono-, di-, and triphosphates. UTP was the predominant early metabolite in Huh-7 cells and primary human hepatocytes, suggesting deamination of NHC as the primary catabolic pathway. The intracellular half-lives of radiolabeled NHC-triphosphate and of CTP and UTP derived from NHC incubation in Huh-7 cells were calculated to be 3.0 ± 1.3, 10.4 ± 3.3, and 13.2 ± 3.5 h (means ± standard deviations), respectively. Studies using monkey and human whole blood demonstrated more-rapid deamination and oxidation in monkey cells than in human cells, suggesting that NHC may not persist long enough in plasma to be delivered to liver cells.

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Blight, Keril J., Jane A. McKeating, and Charles M. Rice. "Highly Permissive Cell Lines for Subgenomic and Genomic Hepatitis C Virus RNA Replication." Journal of Virology 76, no. 24 (December 15, 2002): 13001–14. http://dx.doi.org/10.1128/jvi.76.24.13001-13014.2002.

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ABSTRACT Hepatitis C virus (HCV) replication appears to be restricted to the human hepatoma cell line Huh-7, indicating that a favorable cellular environment exists within these cells. Although adaptive mutations in the HCV nonstructural proteins typically enhance the replicative capacity of subgenomic replicons in Huh-7 cells, replication can only be detected in a subpopulation of these cells. Here we show that self-replicating subgenomic RNA could be eliminated from Huh-7 clones by prolonged treatment with alpha interferon (IFN-α) and that a higher frequency of cured cells could support both subgenomic and full-length HCV replication. The increased permissiveness of one of the cured cell lines allowed us to readily detect HCV RNA and antigens early after RNA transfection, eliminating the need for selection of replication-positive cells. We also demonstrate that a single amino acid substitution in NS5A is sufficient for establishing HCV replication in a majority of cured cells and that the major phosphate acceptor site of subtype 1b NS5A is not essential for HCV replication.

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Osabe, Makoto, and Masahiko Negishi. "Active ERK1/2 Protein Interacts with the Phosphorylated Nuclear Constitutive Active/Androstane Receptor (CAR; NR1I3), Repressing Dephosphorylation and Sequestering CAR in the Cytoplasm." Journal of Biological Chemistry 286, no. 41 (August 26, 2011): 35763–69. http://dx.doi.org/10.1074/jbc.m111.284596.

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The nuclear constitutive active/androstane receptor (CAR) is inactivated and sequestered in the cytoplasm when Thr-38 is phosphorylated. Here, we have demonstrated that activated ERK1/2 interacts with phosphorylated CAR to repress dephosphorylation of Thr-38. The phosphorylation-dependent interaction between CAR and ERK1/2 was examined by co-immunoprecipitation experiments of ectopically expressed FLAG-tagged CAR T38A and CAR T38D mutants with endogenous phospho-ERK1/2 in Huh-7 cells. Phospho-ERK1/2 coprecipitated only the phosphorylation-mimicking CAR T38D mutant; this coprecipitation was mediated by the interaction with the xenochemical response signal peptide near the C terminus of CAR. This interaction increased after EGF treatment and decreased after treatment with the MEK inhibitor U0126 as well as after knockdown of MEK1/2 by shRNA in Huh-7 cells. The phosphorylation levels of Thr-38 of CAR decreased in U0126-treated Huh-7 cells. Thus, activated ERK1/2 interacts with CAR and represses dephosphorylation of Thr-38, providing a cell signal-regulated mechanism for CAR activation.

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Grünvogel, Oliver, Katharina Esser-Nobis, Marc P. Windisch, Michael Frese, Martin Trippler, Ralf Bartenschlager, Volker Lohmann, and Marco Binder. "Type I and type II interferon responses in two human liver cell lines (Huh-7 and HuH6)." Genomics Data 7 (March 2016): 166–70. http://dx.doi.org/10.1016/j.gdata.2015.12.017.

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Nakabayashi, H., T. Hashimoto, Y. Miyao, K. K. Tjong, J. Chan, and T. Tamaoki. "A position-dependent silencer plays a major role in repressing alpha-fetoprotein expression in human hepatoma." Molecular and Cellular Biology 11, no. 12 (December 1991): 5885–93. http://dx.doi.org/10.1128/mcb.11.12.5885-5893.1991.

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A large percentage of human hepatomas produce alpha-fetoprotein (AFP), but the levels of AFP expression vary greatly among hepatomas. To understand the molecular basis for this variation, we analyzed transcriptional regulatory activities associated with the 5'-flanking region of the AFP gene in two human hepatoma cell lines, HuH-7 and huH-1/cl-2, which produce a high and a low level of AFP, respectively. We found that the low level of AFP production in huH-1/cl-2 is due to the action of at least two silencer regions located between the enhancer and the promoter of the AFP gene. In contrast, no silencer activity is expressed in HuH-7. We identified 5'-CTTCATAACTAATACTT-3' to be a core sequence responsible for the negative regulatory activity. This sequence is repeated four times in a strong, distal silencer region, Sd, whereas one copy is present in a weak, proximal silencer region, Sp. The silencer reduces transcriptional initiation by blocking enhancer activation of the AFP promoter in a position-dependent manner. The silencer functions in the presence of positive transcription factors and may play a key role in developmental repression as well as variable expression of the AFP gene in hepatomas.

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Nakabayashi, H., T. Hashimoto, Y. Miyao, K. K. Tjong, J. Chan, and T. Tamaoki. "A position-dependent silencer plays a major role in repressing alpha-fetoprotein expression in human hepatoma." Molecular and Cellular Biology 11, no. 12 (December 1991): 5885–93. http://dx.doi.org/10.1128/mcb.11.12.5885.

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A large percentage of human hepatomas produce alpha-fetoprotein (AFP), but the levels of AFP expression vary greatly among hepatomas. To understand the molecular basis for this variation, we analyzed transcriptional regulatory activities associated with the 5'-flanking region of the AFP gene in two human hepatoma cell lines, HuH-7 and huH-1/cl-2, which produce a high and a low level of AFP, respectively. We found that the low level of AFP production in huH-1/cl-2 is due to the action of at least two silencer regions located between the enhancer and the promoter of the AFP gene. In contrast, no silencer activity is expressed in HuH-7. We identified 5'-CTTCATAACTAATACTT-3' to be a core sequence responsible for the negative regulatory activity. This sequence is repeated four times in a strong, distal silencer region, Sd, whereas one copy is present in a weak, proximal silencer region, Sp. The silencer reduces transcriptional initiation by blocking enhancer activation of the AFP promoter in a position-dependent manner. The silencer functions in the presence of positive transcription factors and may play a key role in developmental repression as well as variable expression of the AFP gene in hepatomas.

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Tabata, Yuki, and Yoshihiro Shidoji. "Hepatic monoamine oxidase B is involved in endogenous geranylgeranoic acid synthesis in mammalian liver cells." Journal of Lipid Research 61, no. 5 (February 24, 2020): 778–89. http://dx.doi.org/10.1194/jlr.ra119000610.

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Geranylgeranoic acid (GGA) originally was identified in some animals and has been developed as an agent for preventing second primary hepatoma. We previously have also identified GGA as an acyclic diterpenoid in some medicinal herbs. Recently, we reported that in human hepatoma-derived HuH-7 cells, GGA is metabolically labeled from 13C-mevalonate. Several cell-free experiments have demonstrated that GGA is synthesized through geranylgeranial by oxygen-dependent oxidation of geranylgeraniol (GGOH), but the exact biochemical events giving rise to GGA in hepatoma cells remain unclear. Monoamine oxidase B (MOAB) has been suggested to be involved in GGOH oxidation. Here, using two human hepatoma cell lines, we investigated whether MAOB contributes to GGA biosynthesis. Using either HuH-7 cell lysates or recombinant human MAOB, we found that: 1) the MAO inhibitor tranylcypromine dose-dependently downregulates endogenous GGA levels in HuH-7 cells; and 2) siRNA-mediated MAOB silencing reduces intracellular GGA levels in HuH-7 and Hep3B cells. Unexpectedly, however, CRISPR/Cas9-generated MAOB-KO human hepatoma Hep3B cells had GGA levels similar to those in MAOB-WT cells. A sensitivity of GGA levels to siRNA-mediated MAOB downregulation was recovered when the MAOB-KO cells were transfected with a MAOB-expression plasmid, suggesting that MAOB is the enzyme primarily responsible for GGOH oxidation and that some other latent metabolic pathways may maintain endogenous GGA levels in the MAOB-KO hepatoma cells. Along with the previous findings, these results provide critical insights into the biological roles of human MAOB and provide evidence that hepatic MAOB is involved in endogenous GGA biosynthesis via GGOH oxidation.

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Chen, Li-Jeng, Tsai-Ching Hsu, Hsiang-Lin Chan, Chiao-Fan Lin, Jing-Yu Huang, Robert Stewart, Bor-Show Tzang, and Vincent Chin-Hung Chen. "Protective Effect of Escitalopram on Hepatocellular Carcinoma by Inducing Autophagy." International Journal of Molecular Sciences 23, no. 16 (August 17, 2022): 9247. http://dx.doi.org/10.3390/ijms23169247.

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Background: Hepatocellular carcinoma (HCC) is an aggressive cancer with poor prognosis. Although recent research has indicated that selective serotonin reuptake inhibitors (SSRIs), including escitalopram, have anticancer effects, little is known about the effects of escitalopram on HCC. Methods: Both in vitro and in vivo studies were conducted to verify the potentials of escitalopram on HCC treatment. To explore whether the effects of escitalopram are clinically consistent with laboratory findings, a nationwide population-based cohort study was also adopted to examine the association between escitalopram and HCC risk. Results: As compared with THLE-3 cells, escitalopram significantly inhibited the proliferation of HepG2 and Huh-7 cells. Specifically, escitalopram significantly induced autophagy in HepG2 and Huh-7 cells by increasing the LC3-II/LC3-I ratio and the expression of ATG-3, ATG-5, ATG-7, and Beclin-1 proteins. Moreover, escitalopram significantly inhibited the growth of xenografted Huh-7 cells in SCID mice that were treated with 12.5 mg/kg escitalopram. Accordingly, the risk of HCC was negatively correlated with escitalopram use. Conclusions: These findings provided evidence supporting the therapeutic potential of escitalopram for HCC. Both laboratory and nationwide population-based cohort evidence demonstrated the attenuated effects of escitalopram on HCC.

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Chuang, Yung Chun, and Trai Ming Yeh. "Macrophage migration inhibitory factor enhances dengue virus replication through autophagy formation and ROS generation (168.10)." Journal of Immunology 188, no. 1_Supplement (May 1, 2012): 168.10. http://dx.doi.org/10.4049/jimmunol.188.supp.168.10.

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Abstract Macrophage migration inhibitory factor (MIF) is a proinflammatory cytokine which has been identified as a risk factor in dengue virus (DENV) infection. Previous study showed MIF deficient mice exhibit lower virus titer than wild type mice during DENV infection. However, the mechanism is still unknown. Herein, we first examined whether MIF interfere DENV replication in vitro. By using flow cytometry assay, we found both MIF inhibitor ISO-1 or MIF knock down could reduce DENV replication in Huh 7 cells. Moreover, we found that reactive oxygen species (ROS) of Huh 7 cells induced by DENV infection were also decreased in the presence of ISO-1. ROS is an inducer of autophagy which has been showed to be involved in DENV replication. Thus, we hypothesize MIF facilitate DENV replication through ROS generation and autophagy formation. We found ISO-1, MIF knock down, or ROS scavenger N-acetyl-L-cysteine (NAC) could reduce autophagy during DENV infection by western blotting analysis. The LC3 punctae formation was also reduced in MIF knock down Huh 7 cells during DENV infection. Taken together, blocking of MIF by its inhibitor or antibody may not only prevent inflammation but also viral replication during DENV infection.

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Yeh, Trai-Ming, and Yung-Chun Chuang. "Macrophage migration inhibitory factor induces autophagy via reactive oxygen species generation (174.10)." Journal of Immunology 188, no. 1_Supplement (May 1, 2012): 174.10. http://dx.doi.org/10.4049/jimmunol.188.supp.174.10.

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Abstract Autophagy is an evolutionarily conserved catabolic process which maintains cellular homeostasis under stress conditions such as starvation and pathogen infection. Macrophage migration inhibitory factor (MIF) is a multifunctional cytokine that plays important roles in inflammation and tumorigenesis. Cytokines such as IL-1β and TNF-α that are induced by MIF have been shown to be involved in the induction of autophagy. However, the actually role of MIF in autophagy is still unclear. Here, we demonstrated that incubation of human hepatoma cell line Huh-7 cells with recombinant MIF (rMIF) induced reactive oxygen species (ROS) production and autophagy formation, including LC3-II expression, LC3 punctae formation and mitochondria membrane potential loss. The autophagy induced by rMIF was inhibited in the presence of MIF inhibitor, ISO-1 as well as ROS scavenger N-acetyl-L-cysteine (NAC). In addition, serum starvation-induced MIF release and autophagy of Huh-7 cells were partly blocked in the presence of NAC. Moreover, diminished MIF expression by shRNA transfection or inhibition of MIF by ISO-1 decreased serum starvation-induced autophagy of Huh-7 cells. Taken together, these data suggest that MIF is involved in autophagy through ROS generation to prevent cell death.

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Breiman, Adrien, Damien Vitour, Myriam Vilasco, Catherine Ottone, Sonia Molina, Lydiane Pichard, Chantal Fournier, et al. "A hepatitis C virus (HCV) NS3/4A protease-dependent strategy for the identification and purification of HCV-infected cells." Journal of General Virology 87, no. 12 (December 1, 2006): 3587–98. http://dx.doi.org/10.1099/vir.0.82214-0.

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As a tool for the identification and/or purification of hepatitis C virus (HCV)-infected cells, a chimeric form of the Gal4VP16 transcription factor was engineered to be activated only in the presence of the HCV NS3/4A protease and to induce different reporter genes [choramphenical acetyltransferase (CAT), green fluorescent protein (GFP) and the cell-surface marker H-2Kk] through the (Gal4)5-E1b promoter. For this, the NS5A/5B trans-cleavage motif of HCV of genotype 1a was inserted between Gal4VP16 and the N terminus of the endoplasmic reticulum (ER)-resident protein PERK, and it was demonstrated that it could be cleaved specifically by NS3/4A. Accordingly, transient transfection in tetracycline-inducible UHCV-11 cells expressing the HCV polyprotein of genotype 1a revealed the migration of the Gal4VP16 moiety of the chimera from the ER to the nucleus upon HCV expression. Activation of the chimera provoked specific gene induction, as shown by CAT assay, first in UHCV-11 cells and then in Huh-7 cells expressing an HCV replicon of genotype 1b (Huh-7 Rep). In addition, the GFP reporter gene allowed rapid fluorescence monitoring of HCV expression in the Huh-7 Rep cells. Finally, the chimera was introduced into Huh-7.5 cells infected with cell culture-generated HCV JFH1 (genotype 2a), allowing the purification of the HCV-infected cells by immunomagnetic cell sorting using H-2Kk as gene reporter. In conclusion, the Gal4VP16 chimera activation system can be used for the rapid identification and purification of HCV-infected cells.

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Al-Majid, Abdullah, Mohammad Islam, Saleh Atef, Fardous El-Senduny, Farid Badria, Yaseen Elshaier, M. Ali, Assem Barakat, and A. Motiur Rahman. "Synthesis of Pyridine-Dicarboxamide-Cyclohexanone Derivatives: Anticancer and α-Glucosidase Inhibitory Activities and In Silico Study." Molecules 24, no. 7 (April 4, 2019): 1332. http://dx.doi.org/10.3390/molecules24071332.

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An efficient and practical method for the synthesis of 2,6-diaryl-4-oxo-N,N′-di(pyridin-2-yl)cyclohexane-1,1-dicarboxamide is described in this present study, which occurs through a double Michael addition reaction between diamide and various dibenzalacetones. The reaction was carried out in dichloromethane (DCM) in the presence of 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU). The anticancer activities of the synthesized compounds were evaluated in several cancer cell lines, including MCF-7, MDA-MB-231, SAS, PC-3, HCT-116, HuH-7 and HepG2 cells. From these experiments, we determined that MDA-MB-231 was the most sensitive cancer cell line to the compounds 3c, 3e, 3d, 3j and 3l, which exhibited variable anticancer activities (3l [IC50 = 5 ± 0.25 µM] > 3e [IC50 = 5 ± 0.5 µM] > 3c [IC50 = 7 ± 1.12 µM] > 3d [IC50 = 18 ± 0.87 µM] > 3j [IC50 = 45 ± 3 µM]). Of these, 3l (substituted p-trifluoromethylphenyl and chloropyridine) showed good potency (IC50 = 6 ± 0.78 µM) against HCT-116 colorectal cancer cells and exhibited high toxicity against HuH-7 liver cancer cells (IC50 = 4.5 ± 0.3 µM). These values were three times higher than the values reported for cisplatin (IC50 of 8 ± 0.76 and 14.7 ± 0.5 µM against HCT-116 and HuH-7 cells, respectively). The highest α-glucosidase inhibitory activity was detected for the 3d, 3i and 3j compounds. The details of the binding mode of the active compounds were clarified by molecular docking studies.

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Kim, Eun-Young, and An-Keun Kim. "Apigenin Sensitizes Huh-7 Human Hepatocellular Carcinoma Cells to TRAIL-induced Apoptosis." Biomolecules and Therapeutics 20, no. 1 (January 31, 2012): 62–67. http://dx.doi.org/10.4062/biomolther.2012.20.1.062.

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Lee, Chang Min, Yong Jun Choi, See-Hyoung Park, and Myeong Jin Nam. "Indole-3-carbinol induces apoptosis in human hepatocellular carcinoma Huh-7 cells." Food and Chemical Toxicology 118 (August 2018): 119–30. http://dx.doi.org/10.1016/j.fct.2018.05.014.

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Eyre, Nicholas S., Renee J. Phillips, Scott Bowden, Evelyn Yip, Ben Dewar, Stephen A. Locarnini, and Michael R. Beard. "Hepatitis B virus and hepatitis C virus interaction in Huh-7 cells." Journal of Hepatology 51, no. 3 (September 2009): 446–57. http://dx.doi.org/10.1016/j.jhep.2009.04.025.

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Dahari, Harel, Ruy M. Ribeiro, Charles M. Rice, and Alan S. Perelson. "Mathematical Modeling of Subgenomic Hepatitis C Virus Replication in Huh-7 Cells." Journal of Virology 81, no. 2 (October 11, 2006): 750–60. http://dx.doi.org/10.1128/jvi.01304-06.

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ABSTRACT Cell-based hepatitis C virus (HCV) replicon systems have provided a means for understanding HCV replication mechanisms and for testing new antiviral agents. We describe here a mathematical model of HCV replication that assumes that the translation of the HCV polyprotein occurs in the cytoplasm, that HCV RNA synthesis occurs in vesicular-membrane structures, and that the strategy of replication involves a double-stranded RNA intermediate. Our results shed light on the intracellular dynamics of subgenomic HCV RNA replication from transfection to steady state within Huh-7 cells. We predict the following: (i) about 6 × 103 ribosomes are involved in generating millions of HCV NS5B-polymerase molecules in a Huh-7 cell, (ii) the observed 10:1 asymmetry of plus- to minus-strand RNA levels can be explained by a higher-affinity (200-fold) interaction of HCV NS5B polymerase-containing replication complexes with HCV minus-strand RNA over HCV plus-strand RNA in order to initiate synthesis, (iii) the latter higher affinity can also account for the observed ∼6:1 plus-strand/minus-strand ratio in vesicular-membrane structures, and (iv) the introduction of higher numbers of HCV plus-strand RNA by transfection leads to faster attainment of steady-state but does not change the steady-state HCV RNA level. Fully permissive HCV replication systems have been developed, and the model presented here is a first step toward building a comprehensive model for complete HCV replication. Moreover, the model can serve as an important tool in understanding HCV replication mechanisms and should prove useful in designing and evaluating new antivirals against HCV.

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Yamamoto, T., Y. Moriwaki, O. Agbedana, S. Takahashi, M. Suda, and K. Higashino. "Oxypurine Metabolism of Xanthine Oxidase-Deficient Hepatoma-Derived Cell Line HuH-7." Hormone and Metabolic Research 26, no. 08 (August 1994): 389–91. http://dx.doi.org/10.1055/s-2007-1001714.

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Mahou, Redouan, Nhu Mai Tran, Murielle Dufresne, Cécile Legallais, and Christine Wandrey. "Encapsulation of Huh-7 cells within alginate-poly(ethylene glycol) hybrid microspheres." Journal of Materials Science: Materials in Medicine 23, no. 1 (December 9, 2011): 171–79. http://dx.doi.org/10.1007/s10856-011-4512-3.

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Lohmann, Volker, Sandra Hoffmann, Ulrike Herian, Francois Penin, and Ralf Bartenschlager. "Viral and Cellular Determinants of Hepatitis C Virus RNA Replication in Cell Culture." Journal of Virology 77, no. 5 (March 1, 2003): 3007–19. http://dx.doi.org/10.1128/jvi.77.5.3007-3019.2003.

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ABSTRACT Studies on the replication of hepatitis C virus (HCV) have been facilitated by the development of selectable subgenomic replicons replicating in the human hepatoma cell line Huh-7 at a surprisingly high level. Analysis of the replicon population in selected cells revealed the occurrence of cell culture-adaptive mutations that enhance RNA replication substantially. To gain a better understanding of HCV cell culture adaptation, we characterized conserved mutations identified by sequence analysis of 26 independent replicon cell clones for their effect on RNA replication. Mutations enhancing replication were found in nearly every nonstructural (NS) protein, and they could be subdivided into at least two groups by their effect on replication efficiency and cooperativity: (i) mutations in NS3 with a low impact on replication but that enhanced replication cooperatively when combined with highly adaptive mutations and (ii) mutations in NS4B, -5A, and -5B, causing a strong increase in replication but being incompatible with each other. In addition to adaptive mutations, we found that the host cell plays an equally important role for efficient RNA replication. We tested several passages of the same Huh-7 cell line and found up to 100-fold differences in their ability to support replicon amplification. These differences were not due to variations in internal ribosome entry site-dependent translation or RNA degradation. In a search for cellular factor(s) that might be responsible for the different levels of permissiveness of Huh-7 cells, we found that replication efficiency decreased with increasing amounts of transfected replicon RNA, indicating that viral RNA or proteins are cytopathic or that host cell factors in Huh-7 cells limit RNA amplification. In summary, these data show that the efficiency of HCV replication in cell culture is determined both by adaptation of the viral sequence and by the host cell itself.

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Xu, Jifan, Bo Du, Yunfeng Liu, and Chonglin Tao. "Magnoflorine promotes Huh-7 cell apoptosis and autophagy by regulating PI3K/Akt/mTOR pathway." Quality Assurance and Safety of Crops & Foods 14, no. 1 (January 20, 2022): 39–45. http://dx.doi.org/10.15586/qas.v14i1.1013.

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Hepatoma is a malignant tumor with high rates of heterogeneity, metastasis, and mortality. Currently, there is no effective treatment available for hepatoma. In order to treat advanced hepatoma in a better manner, new and more effective therapeutic targets still need to be developed. Magnoflorine (MGN) is a quaternary ammonium alkaloid with a variety of therapeutic properties. MGN inhibited the proliferation of lung cancer, breast cancer, glioma, and rhabdomyosarcoma cells, induced apoptosis, and blocked cell cycle. However, its possible effects on the progression of hepatoma are still indefinite. In this study, the effects of MGN on the progression of hepatoma in vitro and the underlying mechanisms were determined. MGN suppressed the proliferation, induced the autophagy, and stimulated the apoptosis of human hepatoma Huh-7 cells. Mechanically, MGN could regulate PI3K/AKT/mTOR pathway, which therefore affects the progression of hepatoma in vitro. Taken together, MGN affected Huh-7 cell proliferation, autophagy, and apoptosis, and might act as a promising therapeutic drug for treating hepatoma.

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Windisch, Marc P., Michael Frese, Artur Kaul, Martin Trippler, Volker Lohmann, and Ralf Bartenschlager. "Dissecting the Interferon-Induced Inhibition of Hepatitis C Virus Replication by Using a Novel Host Cell Line." Journal of Virology 79, no. 21 (November 1, 2005): 13778–93. http://dx.doi.org/10.1128/jvi.79.21.13778-13793.2005.

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ABSTRACT The Hepatitis C virus (HCV), a member of the family Flaviviridae, is a major cause of chronic liver disease. Patients are currently treated with alpha interferon (IFN-α) that is given alone or in combination with ribavirin. Unfortunately, this treatment is ineffective in eliminating the virus in a large proportion of individuals. IFN-induced antiviral activities have been intensively studied in the HCV replicon system. It was found that both IFN-α and IFN-γ inhibit HCV replicons, but the underlying mechanisms have not yet been identified. Of note is that nearly all of these studies were performed with the human hepatoma cell line Huh-7. Here, we report that genotypes 1b and 2a replicons also replicate in the human hepatoblastoma cell line HuH6. Similar to what has been described for Huh-7 cells, we observed that efficient HCV replication in HuH6 cells depends on the presence of cell culture-adaptive mutations and the permissiveness of the host cell. However, three major differences exist: in HuH6 cells, viral replication is (i) independent from ongoing cell proliferation, (ii) less sensitive to certain antiviral compounds, and (iii) highly resistant to IFN-γ. The latter is not due to a general defect in IFN signaling, as IFN-γ induces the nuclear translocation of signal transducer and activator of transcription 1 (STAT1), the enhanced transcription of several IFN-regulated genes, and the inhibition of unrelated viruses such as influenza A virus and Semliki Forest virus. Taken together, the results establish HuH6 replicon cells as a valuable tool for IFN studies and for the evaluation of antiviral compounds.

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Journal articles on the topic 'HuH-7'

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