parental hepg2 human hepatoma cells  (ATCC)

Journal: Journal of Ginseng Research

Article Title: Ginsenoside Rg3 in combination with artesunate overcomes sorafenib resistance in hepatoma cell and mouse models

doi: 10.1016/j.jgr.2021.07.002

Figure Lengend Snippet: Rg3-plus-ART reduces viability of, and induced apoptosis in HepG2-SR cells. (A) - (B) Chemical structures of Rg3 and ART. (C) Viability of HepG2-SR cells. The viability of solvent-treated cells was regarded as 100%. Data from three independent experiments are presented as mean ± SD. ∗ P < 0.05, ∗∗ P < 0.01 vs. the solvent-treated (48 hrs) group; # P < 0.05, ## P < 0.01 vs. the solvent-treated (72 hrs) group. § CDI <1, §§ CDI <0.7. (D) Rg3-plus-ART induces apoptosis in HepG2-SR cells. Representative scatter graphs are shown in the upper panels. FITC positive cells (the right quadrants) are regarded as apoptotic cells. Quantitative results are shown in the lower panel. Data from three independent experiments are presented as mean ± SD. ∗ P < 0.05, ∗∗ P < 0.01 vs. the solvent-treated group; # P < 0.05 vs. the 50 μM Rg3 + 10 μM ART-treated group; Δ P < 0.05 vs. the 75 μM Rg3 + 15 μM ART-treated group. (E) Protein levels of cleaved-PARP in HepG2-SR cultures. Cells were treated with indicated concentrations of Rg3-plus-ART for 48 hrs. β-Actin served as a loading control. Representative immunoblotting results are shown in the left panel, and quantitative results are shown in the right panel. Data are expressed as mean ± SD of 3 independent experiments. ∗ P < 0.05, ∗∗ P < 0.01 vs. the solvent-treated group. In (C) - (E) , brivanib was used as positive control.

Article Snippet: Briefly, parental HepG2 human hepatoma cells (purchased from ATCC) were cultured in DMEM with 5% FBS and increasing concentrations of sorafenib (0.1–5 μM) for 3 months and then maintained in 5 μM sorafenib thereafter.

Techniques: Solvent, Control, Western Blot, Positive Control

Journal: Journal of Ginseng Research

Article Title: Ginsenoside Rg3 in combination with artesunate overcomes sorafenib resistance in hepatoma cell and mouse models

doi: 10.1016/j.jgr.2021.07.002

Figure Lengend Snippet: Rg3-plus-ART suppresses HepG2-SR tumor growth in mice. (A) Images of mice after drug administration. (B) Tumor weights of mice at the end of the experiment. Representative images of tumors (the left panel) and weights of tumors (the right panel) are shown. (C) Tumor volumes. Tumor volumes of each mouse were measured at the indicated time points. (D) Body weights of mice at the indicated time points. Data are expressed as mean ± SD (n = 6). ∗ P < 0.05, ∗∗ P < 0.01 vs. the model group.

Article Snippet: Briefly, parental HepG2 human hepatoma cells (purchased from ATCC) were cultured in DMEM with 5% FBS and increasing concentrations of sorafenib (0.1–5 μM) for 3 months and then maintained in 5 μM sorafenib thereafter.

Techniques:

Journal: Journal of Ginseng Research

Article Title: Ginsenoside Rg3 in combination with artesunate overcomes sorafenib resistance in hepatoma cell and mouse models

doi: 10.1016/j.jgr.2021.07.002

Figure Lengend Snippet: Rg3-plus-ART inhibits the Src/STAT3 pathway in tumor tissues and in cultured HepG2-SR cells . (A) Protein levels of phospho-Src (Tyr416), phospho-STAT3 (Tyr705), Src and STAT3 in tumor tissues of mice. Data are expressed as mean ± SD (n = 6) ∗ P < 0.05, ∗∗ P < 0.01 vs. the model group. (B) Protein levels of phospho-Src (Tyr416), phospho-STAT3 (Tyr705), Src and STAT3 in HepG2-SR cultures. (C) Protein levels of Bcl-2 and Mcl-1 in HepG2-SR cultures. In (B) and (C) , cells were incubated with the indicated concentrations of Rg3-plus-ART or stattic for 24 hrs. In (A) - (C) , β-actin served as the loading control. (D) Protein levels of STAT3 in cytoplasm and nuclear extracts. Lamin B1 and β-actin served as the loading controls of nuclear and cytoplasmic extracts, respectively. Cells were incubated with the indicated concentrations of Rg3-plus-ART or stattic for 30 min. Representative immunoblotting results are shown in the left panel, and quantitative results are shown in the right panel. In (B) - (D) , Data are expressed as mean ± SD of 3 independent experiments. ∗ P < 0.05, ∗∗ P < 0.01 vs. the solvent-treated group.

Article Snippet: Briefly, parental HepG2 human hepatoma cells (purchased from ATCC) were cultured in DMEM with 5% FBS and increasing concentrations of sorafenib (0.1–5 μM) for 3 months and then maintained in 5 μM sorafenib thereafter.

Techniques: Cell Culture, Incubation, Control, Western Blot, Solvent

Journal: Journal of Ginseng Research

Article Title: Ginsenoside Rg3 in combination with artesunate overcomes sorafenib resistance in hepatoma cell and mouse models

doi: 10.1016/j.jgr.2021.07.002

Figure Lengend Snippet: Over-activation of STAT3 diminishes the inhibitory effects of Rg3-plus-ART on cell viability in HepG2-SR cells. HepG2-SR cells were transduced with the Ad-Empty vector (HepG2-SR Empty vector ) or the Ad-STAT3C plasmid (HepG2-SR STAT3C ). (A) GFP expression and the protein levels of STAT3, phospho-STAT3 (Tyr 705) and Flag in HepG2-SR Empty vector and HepG2-SR STAT3C cells. Representative green fluorescent and bright-field microscopy images of HepG2-SR Empty vector and HepG2-SR STAT3C (the left panel, scale bar, 10 μm), and representative immunoblotting results (the right panel) are shown. (B) Over-activation of STAT3 diminishes the anti-proliferative effects of Rg3-plus-ART in HepG2-SR cells. Differences of relative cell viabilities between Rg3-plus-ART-treated HepG2-SR Empty vector cells and Rg3-plus-ART-treated HepG2-SR STAT3C cells were calculated (n = 3). ∗ P < 0.05, ∗∗ P < 0.01.

Article Snippet: Briefly, parental HepG2 human hepatoma cells (purchased from ATCC) were cultured in DMEM with 5% FBS and increasing concentrations of sorafenib (0.1–5 μM) for 3 months and then maintained in 5 μM sorafenib thereafter.

Techniques: Activation Assay, Transduction, Plasmid Preparation, Expressing, Microscopy, Western Blot

Journal: Journal of Ginseng Research

Article Title: Ginsenoside Rg3 in combination with artesunate overcomes sorafenib resistance in hepatoma cell and mouse models

doi: 10.1016/j.jgr.2021.07.002

Figure Lengend Snippet: Induction of ROS production contributes to Rg3-plus-ART's inhibitory effects on cell viability and STAT3 activation in HepG2-SR cultures. (A) Rg3, ART and Rg3-plus-ART induced ROS production in HepG2-SR cells. Relative ROS production in the control group at 0 hr was regarded as 1. Data are expressed as mean ± SD of 3 independent experiments. ∗ P < 0.05, ∗∗ P < 0.01 vs. the solvent-treated control group. (B) Fluorescence microscopy images of DAPI- and DCF-DA-stained HepG2-SR cells treated with Rg3-plus-ART and/or NAC for 8 hrs. ROS generated in cells displayed green fluorescence, and cell nuclei displayed blue fluorescence. Scale bar, 10 μm. (C) Co-treatment with NAC diminishes Rg3-plus-ART's inhibitory effects on the viability of HepG2-SR cells. Cells were treated with indicated concentrations of Rg3-plus-ART with or without 5 mM NAC for 72 hrs. Differences in the relative cell viabilities between the NAC-treated group and NAC-untreated group were calculated. Data from three independent experiments are presented as mean ± SD. ∗ P < 0.05, ∗∗ P < 0.01. (D) NAC diminishes the inhibitory effects of Rg3-plus-ART on STAT3 phosphorylation in HepG2-SR cultures. Cells were treated with 75 μM Rg3 + 15 μM ART and/or 5 mM NAC for 24 hrs. Representative immunoblotting results are shown in the left panel, and quantitative results are shown in the right panel. Differences in the relative protein levels of phospho-STAT3 (Tyr705) between the Rg3-plus-ART-treated group and Rg3-plus-ART-untreated group were calculated. Data are expressed as mean ± SD of 3 independent experiments. ∗ P < 0.05, ∗∗ P < 0.01.

Article Snippet: Briefly, parental HepG2 human hepatoma cells (purchased from ATCC) were cultured in DMEM with 5% FBS and increasing concentrations of sorafenib (0.1–5 μM) for 3 months and then maintained in 5 μM sorafenib thereafter.

Techniques: Activation Assay, Control, Solvent, Fluorescence, Microscopy, Staining, Generated, Phospho-proteomics, Western Blot