normal human liver cell line wrl-68  (National Centre for Cell Science)

Journal: International Journal of Molecular Sciences

Article Title: Protective Effect of Antrodia cinnamomea Extract against Irradiation-Induced Acute Hepatitis

doi: 10.3390/ijms20040846

Figure Lengend Snippet: The protective effects of Antrodia cinnamomea extract (ACE) against irradiation in human normal liver CL48 cells. ( A ) Cytotoxicity of irradiation in CL48. Cells were treated with different dosages (0, 5, 10, 15, and 20 Gy) of irradiation and counted by hemocytometer at intervals as indicated. ( B ) Determination of the optimal ACE dosages. Cells were treated with different concentrations of ACE for 24 h or 48 h, followed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay for cell survival. For the control group, the same concentrations of ethanol were used for each ACE treatment experiment. ( C ) The protective effect of ACE against irradiation. Cells were pre-treated with different concentrations of ACE for 16 h followed by various dosages of irradiation; they were evaluated by MTT assay 48 h later. Cell survival was evaluated by MTT assay. Results were obtained from three independent experiments; each experiment was done in triplicate. **, p < 0.01, as compared with the control group.

Article Snippet: Normal human embryonic liver cell line CL48 cells (American Type Culture Collection) were cultured in high glucose DMEM with 10% fetal bovine serum (FBS), 0.1 mg/mL of streptomycin, and 100 units/mL of penicillin, at 37 °C in a humidified atmosphere containing 5% CO 2 .

Techniques: Irradiation, MTT Assay, Control

Journal: International Journal of Molecular Sciences

Article Title: Protective Effect of Antrodia cinnamomea Extract against Irradiation-Induced Acute Hepatitis

doi: 10.3390/ijms20040846

Figure Lengend Snippet: Protective effects of ACE against irradiation-induced apoptosis. CL48 cells were pre-treated with various concentrations of ACE for 16 h followed by irradiation at 15 Gy. Cells were harvested 48 h later for determination of caspase-3 activity ( A ), caspase-8 activity ( B ), and caspase-9 activity ( C ). The fold differences of the three caspase activities relative to individual control groups were also presented ( D ). For apoptosis analysis, cells were harvested 30 h after treatment followed by staining with PI and Annexin V ( E ). Results are representative of three independent experiments. The statistical analysis of the percentages of early plus late apoptotic cells ( F ). *, p < 0.05; **, p < 0.01, as compared with the irradiated ACE 0 μg/mL control group.

Article Snippet: Normal human embryonic liver cell line CL48 cells (American Type Culture Collection) were cultured in high glucose DMEM with 10% fetal bovine serum (FBS), 0.1 mg/mL of streptomycin, and 100 units/mL of penicillin, at 37 °C in a humidified atmosphere containing 5% CO 2 .

Techniques: Irradiation, Activity Assay, Control, Staining

Journal: International Journal of Molecular Sciences

Article Title: Protective Effect of Antrodia cinnamomea Extract against Irradiation-Induced Acute Hepatitis

doi: 10.3390/ijms20040846

Figure Lengend Snippet: Reactive oxygen species (ROS) scavenging activity of ACE in CL48 cells. After being treated and harvested as described in the Materials and Methods section, cells were analyzed by flowcytometry after incubation with 2′,7′-dichlorofluorescin diacetate (DCFH-DA) for 15 min. ( A ) Control cells without treatment, ( B ) IR treated only, ( C ) H 2 O 2 treated only, ( D ) pre-treated with NAC before IR, ( E ) pre-treated with ACE 40 μg/mL before IR, ( F ) pre-treated with ACE 80 μg/mL before IR. ( G ) Histograms of B, E, and F were combined to show the dose-dependent ROS scavenging activity of ACE. ( H ) Fold change of control fluorescence intensity by 40 μg/mL and 80 μg/mL of ACE, respectively.

Article Snippet: Normal human embryonic liver cell line CL48 cells (American Type Culture Collection) were cultured in high glucose DMEM with 10% fetal bovine serum (FBS), 0.1 mg/mL of streptomycin, and 100 units/mL of penicillin, at 37 °C in a humidified atmosphere containing 5% CO 2 .

Techniques: Activity Assay, Incubation, Control, Fluorescence

Journal: International Journal of Molecular Sciences

Article Title: Protective Effect of Antrodia cinnamomea Extract against Irradiation-Induced Acute Hepatitis

doi: 10.3390/ijms20040846

Figure Lengend Snippet: Redox-related enzymes expression and activity profiles of CL48 hepatocytes after IR and/or ACE treatments. With or without ACE pre-treatment for 16 h, cells were irradiated with a dose of 15 Gy and harvested 24 h later for RT-PCR ( A ), Western blot ( B ), total superoxide dismutase (SOD) activity ( C ), and glutathione peroxidase (GPx) activity ( D ) analyses. Results were obtained from three independent experiments, each experiment was done in triplicate. **, p < 0.01, as compared with the control IR only group.

Article Snippet: Normal human embryonic liver cell line CL48 cells (American Type Culture Collection) were cultured in high glucose DMEM with 10% fetal bovine serum (FBS), 0.1 mg/mL of streptomycin, and 100 units/mL of penicillin, at 37 °C in a humidified atmosphere containing 5% CO 2 .

Techniques: Expressing, Activity Assay, Irradiation, Reverse Transcription Polymerase Chain Reaction, Western Blot, Control

Journal: International Journal of Molecular Sciences

Article Title: Protective Effect of Antrodia cinnamomea Extract against Irradiation-Induced Acute Hepatitis

doi: 10.3390/ijms20040846

Figure Lengend Snippet: Enhancement of the irradiation-induced nuclear factor erythroid-2-related factor (Nrf2) expression and nuclear translocation by ACE treatment. With or without ACE pre-treatment for 16 h, CL48 cells were irradiated with a dose of 15 Gy and harvested 24 h later for Western blot ( A ) and immunofluorescence staining ( B , left) of Nrf2. The slides were counterstained with 4′,6-diamidino-2-phenylindole (DAPI) (300 nM in PBS) ( B , middle) and merged images ( B , right). ( C ) The nuclear Nrf2 positivity rates were plotted. Results were obtained from random fields of three independent experiments for each group. *, p < 0.05; **, p < 0.01, as compared with the IR only group.

Article Snippet: Normal human embryonic liver cell line CL48 cells (American Type Culture Collection) were cultured in high glucose DMEM with 10% fetal bovine serum (FBS), 0.1 mg/mL of streptomycin, and 100 units/mL of penicillin, at 37 °C in a humidified atmosphere containing 5% CO 2 .

Techniques: Irradiation, Expressing, Translocation Assay, Western Blot, Immunofluorescence, Staining

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Combined therapy with oncolytic adenoviruses encoding TRAIL and IL-12 genes markedly suppressed human hepatocellular carcinoma both in vitro and in an orthotopic transplanted mouse model

doi: 10.1186/s13046-016-0353-8

Figure Lengend Snippet: In vitro effects of Ad-ΔB and Ad-ΔB/TRAIL+Ad-ΔB/IL-12 on Hep3B and HuH7 human HCC cells and normal human liver (WRL68) cells. a Quantitative results of MTT assay showing the inhibitor effects on viability of Hep3B cells at 5 and 10 MOI (the multiplicity of infection) per virus; ( b ) Quantitative results of MTT assay showing the inhibitor effects on viability of HuH7 cells at 10 and 20 MOI per virus; and ( c ) Quantitative results of MTT assay showing the inhibitor effects on viability of WRL68 cells at 50 MOI per virus. Data are represented as mean ± SE. d Representatives of semi-quantitative assessment of cytotoxic potency by crystal violet cytopathic effect assay on Hep3B cells (left panel), HuH7 cells (middle panel), and WRL68 cells (right panel) at indicated MOIs. Ad-ΔB = non-armed control oncolytic adenovirus; Ad-ΔB/TRAIL+Ad-ΔB/IL-12 (or Ad-ΔB/TRAIL+IL-12) = combined therapy with two oncolytic adenoviruses encoding human TRAIL and IL-12 genes, respectively

Article Snippet: The Hep3B human HCC cell line, the WRL68 human normal liver cell line, and the HEK293 human embryonic kidney cell line expressing the E1A region of Ad5 were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA), while the HuH7 human HCC cell line was obtained from Japan Health Science Research Resources (JCRB Genebank, Osaka, Japan).

Techniques: In Vitro, MTT Assay, Infection, Virus, Control