human hepatoma cell line plc prf 5  (ATCC)

Journal: Hepatology Communications

Article Title: Molnupiravir is effective against hepatitis E virus infection in an animal model

doi: 10.1097/HC9.0000000000000944

Figure Lengend Snippet: In vitro evaluation of molnupiravir in HEV cell culture models. (A) Cytotoxicity of molnupiravir, ribavirin, and sofosbuvir in PLC/PRF/5 cells and PRH. Three replicates per concentration. Bars represent the mean and SEM. Mean viability at each concentration was compared with the no-drug control group using the Student t test; ** indicates statistically significant difference ( p <0.05). (B) PRH infected with rHEV and treated with either PBS, varying concentrations of molnupiravir, ribavirin, and sofosbuvir, as well as combinations. (C) Immunofluorescence staining for rHEV-infected PRH with/without antivirals: uninfected cells (negative control), rHEV-infected cells treated with either PBS (no drug control), sofosbuvir 10 μM, ribavirin 200 μM, or molnupiravir 100 μM. Cells were stained with polyclonal anti-rHEV sera and counterstained with DAPI. (D) PLC/PRF/5 cells infected with HEV infectious clone Kernow-p6, (E) PLC/PRF/5 cells infected with human-derived HEV. Cells were treated with either PBS, varying concentrations of molnupiravir, ribavirin, and sofosbuvir, as well as combinations. Error bars represent SEM (n=3 replicates per concentration). Mean viral loads at each concentration were compared with the no-drug control group using the Student t test; * indicates statistically significant difference ( p <0.05) from the untreated mean. ** indicates an extremely statistically significant difference ( p <0.01) from the untreated mean. Abbreviations: HEV, hepatitis E virus; PRH, primary rat hepatocytes; SEM, standard error of the mean; PBS, phosphate-buffered saline; rHEV, Rocahepevirus ratti genotype 1; DAPI, 4′,6-diamidino-2-phenylindole.

Article Snippet: Human hepatoma cell line PLC/PRF/5 (ATCC CRL-8024, LGC Standards, Wesel, Germany) and primary rat hepatocytes (PRH) were used in this study.

Techniques: In Vitro, Cell Culture, Concentration Assay, Control, Infection, Immunofluorescence, Staining, Negative Control, Derivative Assay, Virus, Saline

Journal: Cell Reports Medicine

Article Title: Histone methyl-transferase G9a inhibition boosts the efficacy of immune checkpoint inhibitors in experimental hepatocellular carcinoma

doi: 10.1016/j.xcrm.2026.102717

Figure Lengend Snippet: Antitumoral effects of the G9a-specific inhibitor EZM8266 (A) Colony formation assays performed in PLC/PRF/5 human HCC cells treated with the indicated concentrations of EZM8266 ( n = 3). (B) Anchorage-independent growth assay performed in PLC/PRF/5 cells treated with the indicated concentrations of EZM8266. (C and D) Cell migration (C) and cell invasion (D) assays performed in HuH7 cells treated with the indicated concentrations of EZM8266 (48 h pretreatment and 24 h of transwell migration/invasion) ( n = 3). (E) Volcano plot representing all the genes with significant differential expression ( p < 0.05) in PLC/PRF/5 HCC cells treated with EZM8266 (5 μM for 48 h) vs . control cells and most relevant categories of differentially expressed genes identified by Gene Ontology Biological Process (GO-BP) functional classification. (F) GSEA analysis of specific categories including heatmaps with a list of genes modulated by EZM8266 selected from the RNA-seq data. (G) Effect of EZM8266, IFN-γ, and their combination on the expression of selected genes in the murine HCC cell line PM299L ( n = 3). (H) Validation of the effects of EZM8266, IFN-γ, and their combination, as indicated above, on the expression of selected genes identified in the RNA-seq analyses in the murine HCC cell line PM299L ( n = 3). (I) Experimental protocol for the study of the antitumoral effects of EZM8266 in an orthotopic model of HCC. Representative images of tumors and tumor burden (liver index) in vehicle and EZM8266-treated mice ( n = 9/group). (J) Representative images showing the immunohistochemical detection of CD4 + and CD8 + T cells and quantification of tumor-infiltrating CD8 + and CD4 + T cells (yellow arrows) in vehicle- and EZM8266-treated mice. Scale bars, 100 μm, Data are represented as mean ± SEM. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001 . All the replicates represent biological replicates.

Article Snippet: Human: PLC/PRF/5 , ATCC , Cat# CRL-8024 TM.

Techniques: Growth Assay, Migration, Quantitative Proteomics, Control, Functional Assay, RNA Sequencing, Expressing, Biomarker Discovery, Immunohistochemical staining

Journal: Frontiers in Physiology

Article Title: Long noncoding RNA X-inactive-specific transcript promotes hepatic fibrosis by suppressing ferroptosis in hepatic stellate cells via the miR-663a/GPX4 axis

doi: 10.3389/fphys.2025.1734886

Figure Lengend Snippet: Ethanol induced lncRNA-XIST expression, and silencing of lncRNA-XIST promoted ferroptosis to repress HSC activation. LX-2 cells were treated with 0 mM, 50 mM, 100 mM, and 150 mM ethanol for 24 h or with 100 mM ethanol for 0 h, 12 h, 24 h, and 48 h, respectively, to induce HSC activation. (A) CCK-8 assay to evaluate cell viability; (B) Western blot to determine α-SMA and CoL1A1 expression; (C) RT-qPCR to determine lncRNA-XIST expression; (D) RT-qPCR to measure the lncRNA-XIST level in activated LX-2 cells and hepatocellular carcinoma cells (HepG2, Huh7, and PLC/PRF/5). LX-2 cells were treated with 100 mM ethanol for 24 h, and (E) RT-qPCR to determine lncRNA-XIST mRNA expression; (F) CCK-8 to evaluate cell viability; (G) Colony formation assay to assess cell proliferation; (H) Western blot to measure α-SMA and CoL1A1 levels; (I) BODIPY 581/591 C11 immunofluorescence to observe LPO levels (for cells without LPO, the BODIPY 581/591 C11 fluorescent probe showed orange-red fluorescence; for cells with LPO, the BODIPY 581/591 C11 fluorescent probe exhibited green fluorescence); (J) Kits to determine Fe 2+ , MDA and GSH levels; (K) LDH assay to assess cell death. The cellular experiments were repeated three times independently and the data were depicted as mean ± SD. Data comparisons among multiple groups were performed by one-way ANOVA, with post hoc analysis using Tukey’s multiple comparison test. Differences were considered statistically significant at P < 0.05. The actual difference between the two groups of data was assessed using the effect size Cohen’s d (d) (LX-2 group vs . Con group; LX-2 + si-XIST group vs . LX-2 + si-NC group), with d > 0.8 considered a large effect size.

Article Snippet: Human hepatocellular carcinoma cell lines (HepG2, Huh7, and PLC/PRF/5) purchased from ATCC (Manassas, VA, United States) were cultured in the Dulbecco’s modified Eagle medium supplemented with 10% fetal bovine serum, penicillin (100 IU/mL), and streptomycin (100 mg/mL) [all obtained from Gibco (Grand Island, NY, United States)] and placed in an incubator at 37 °C with 5% CO 2 and 95% humidity.

Techniques: Expressing, Activation Assay, CCK-8 Assay, Western Blot, Quantitative RT-PCR, Colony Assay, Immunofluorescence, Fluorescence, Lactate Dehydrogenase Assay, Comparison