Journal: Journal of cellular and molecular medicine

Article Title: Functional analysis of microRNAs in human hepatocellular cancer stem cells.

doi: 10.1111/j.1582-4934.2011.01282.x

Figure Lengend Snippet: Fig. 1 Characterization of Cancer Stem Cells from human HCC tissues. (A) Real-time analysis of Oct 4, CD 133, Nestin, telom- erase, SSEA-4, AFP and CEA mRNA expres- sion in HCC stem cell under self-renewal con- dition (HSC-SR), under differentiation (HSC- DF) and HepG2 HCC cells. All the markers are significantly up-regulated in HCC cancer stem cells. *P 0.05 when compared with HepG2 group. (B) FACS analysis of HCC stem cells on day 1 before cell sorting. Results are given as the percentage of Oct-4 and CD133

Article Snippet: Human hepatocellular CSCs and parental HCC cells were provided by Celprogen Inc. (San Pedro, CA, USA).

Techniques: FACS

Journal: Journal of cellular and molecular medicine

Article Title: Functional analysis of microRNAs in human hepatocellular cancer stem cells.

doi: 10.1111/j.1582-4934.2011.01282.x

Figure Lengend Snippet: Fig. 4 HCC stem cells are resistant to con- ventional chemotherapy. (A, C) After serum starvation of cultured cells overnight, sorafenib and doxorubicin were added at various concentrations (10 3–10 10 M) and cell viability was assessed after 72 hrs. IC50 graph of high-content image analysis of human hepatocytes and HCC stem cells treated by sorafenib and doxorubicin was illustrated. Quantitative data of MTS assay in the sorafenib treated cells were calcu- lated for IC50 with XLfit software, which is marked in the middle of the box. (B, D) IC50 of sorafenib and doxorubicin in hepato- cytes and HCC stem cells is illustrated in bar graph. HCC stem cells under either self- renewal (HSC-SR) or differentiation (HSC- DF) are more resistance to sorafenib and doxorubicin than HepG2 cells and normal liver stem cells. IC50 results are expressed as the mean S.E. of eight different exper- iments. (E) Groups of five SCID Berge mice were selected 8–10 weeks old per treat- ment group. The human HSC-SRs (1000) as well as parental HCC cells (5 106) were injected subcutaneously and allowed to engraft for 10 days. At day 10 the tumour was visible, and then the doxorubicin treat- ment was started and the mice received the desired concentration of the drug intraper- oteneal three times per week for 14 days. At the end of the 14 days the tumour was dis- sociated into single cell suspension and assayed with Almar Blue to determine per- cent inhibition of the drug. (F, G) Differential expression of CD133 and let-7a in vivo following treatment with doxorubicin by real-time PCR is shown. Data represent percentage change in expression in doxo- cibicin-treated tumours compared with controls. *P 0.05 relative to controls.

Article Snippet: Human hepatocellular CSCs and parental HCC cells were provided by Celprogen Inc. (San Pedro, CA, USA).

Techniques: Cell Culture, MTS Assay, Software, Injection, Concentration Assay, Suspension, Inhibition, Quantitative Proteomics, In Vivo, Real-time Polymerase Chain Reaction, Expressing

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