Journal: Journal of cellular physiology

Article Title: Involvement of Notch-1 in Resistance to Regorafenib in Colon Cancer Cells.

doi: 10.1002/jcp.25206

Figure Lengend Snippet: Fig. 1. Chemical structure of regorafenib.

Article Snippet: Regorafenib were purchased from Selleck Chemicals (Houston, TX) (Stivarga, 2013) (Fig. 1).

Techniques:

Journal: Journal of cellular physiology

Article Title: Involvement of Notch-1 in Resistance to Regorafenib in Colon Cancer Cells.

doi: 10.1002/jcp.25206

Figure Lengend Snippet: Fig. 2. Growth profile of regorafenib-resistant cells. (A) Regorafenib-resistant cells (Reg-R-SW480 cells) and their corresponding parental SW480 cells controls were treated with increasing amounts of regorafenib. Following 24 h incubation, the numbers of viable cells in each well were determined by a proliferation assay as described in “Materials and Methods.” Results were expressed as the percentage of cell growth relative to controls. Each point represents mean SD of three determinations. (B) Cell cycle analysis of Reg-R-SW480 and parental cells. The percentage of cells at each stage of the cell cycle was analyzed by flow cytometry after DNA staining with propidium iodide. Data from a representative experiment (from a total of three) are shown. (C) Values in B represent the percentage of cells undergoing G0-G1 phase arrest. The difference in the percentage of cells in G0-G1 phase arrest between the Reg-R-SW480 and SW480 cells treated both with regorafenib was statistically significant in (P < 0.05, P < 0.01, and P < 0.001. (D) Regorafenib-resistant cells (Reg-R-SW480 cells) maintain resistant phenotype after long–term culture in drug free media. Anti-proliferative effects of Regorafenib were examined in SW480 parental and resistant cultured either in maximal tolerable doses of inhibitor or in drug free conditions for 9 months (Reg-R Drug Free). Colon cancer cells were exposed to the indicated doses of regorafenib for 72 h. The number of viable cells in each well was determined by proliferation assay as described in “Materials and Methods.” Results were expressed as the percentage of cell growth relative to controls. Each point represents mean SD of three determinations. Similar results were obtained in replicate experiments.

Article Snippet: Regorafenib were purchased from Selleck Chemicals (Houston, TX) (Stivarga, 2013) (Fig. 1).

Techniques: Incubation, Proliferation Assay, Cell Cycle Assay, Flow Cytometry, Staining, Cell Culture

Journal: Journal of cellular physiology

Article Title: Involvement of Notch-1 in Resistance to Regorafenib in Colon Cancer Cells.

doi: 10.1002/jcp.25206

Figure Lengend Snippet: Fig. 3. Effect of regorafenib on activation of c-Kit and PDGFR-b in resistant cells (Reg-R SW480 cells) and their corresponding parental SW480 cells controls. (A) Both cell lines were exposed to regorafenib 1 mM for 24 h, then processed for Western blot analysis of p-c-KIT, c-KIT, p-PDGFR-b, PDGFR-b. b-actin was used as a control for protein loading. Blots are representative at of at least three separate experiments. (B) Values represent the mean relative ratios SD from three experiments. P < 0.05, P < 0.01 refers to Reg-R SW480 and parental SW480 treated with regorafenib. (C) Both cell lines were exposed to regorafenib for 24 h, then processed for Western blot analysis of pS473-AKT, AKT, p-ERK1/2, EK1/2, p-mTOR, mTOR. b-actin was used as a control for protein loading. Blots are representative at of at least three separate experiments. (D) Values represent the mean relative ratios SD from three experiments. P < 0.05, P < 0.01 refers to Reg-R SW480 parental SW480 treated with regorafenib. (E) Values represent the mean relative ratios SD from three experiments. P < 0.05, P < 0.01 refers to Reg-R SW480 parental SW480 treated with regorafenib.

Article Snippet: Regorafenib were purchased from Selleck Chemicals (Houston, TX) (Stivarga, 2013) (Fig. 1).

Techniques: Activation Assay, Western Blot, Control

Journal: Journal of cellular physiology

Article Title: Involvement of Notch-1 in Resistance to Regorafenib in Colon Cancer Cells.

doi: 10.1002/jcp.25206

Figure Lengend Snippet: Fig. 4. Effect of regorafenib on expression of c-Kit and Notch-1 in SW480 cells, Reg-R-SW480 cells, Reg-R-SW480-SiRNA-Notch-1 cells, and Reg-R-SW480-SiRNA-control cells. (A) Both cell lines were exposed to regorafenib 1 mM for 24 h, then processed for RT-PCR assay of p-c-KIT and Notch-1. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH)-actin was used as a control. Gels are representative at of at least three separate experiments. (B) Values represent the mean relative ratios SD from three experiments. P < 0.05, P < 0.01 refers to Reg-R SW480 parental SW480 treated with regorafenib. (C) All cell lines (SW480 cells, Reg-R-SW480 cells, Reg-R-SW480-SiRNA-Notch-1 cells, and Reg-R-SW480-SiRNA-control cells) were processed for Western blot analysis Notch-1. b-actin was used as a control for protein loading. Blots are representative at of at least three separate experiments. (D) Values represent the mean relative ratios SD from three experiments. P < 0.05, P < 0.01 refers to Reg-R SW480 cells, parental SW480 cells, Reg-R-SW480-SiRNA-Notch-1 cells, and Reg-R-SW480-SiRNA- control cells.

Article Snippet: Regorafenib were purchased from Selleck Chemicals (Houston, TX) (Stivarga, 2013) (Fig. 1).

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

Journal: Journal of cellular physiology

Article Title: Involvement of Notch-1 in Resistance to Regorafenib in Colon Cancer Cells.

doi: 10.1002/jcp.25206

Figure Lengend Snippet: Fig. 5. Growth profile and effect of regorafenib on expression p-AKT and AKT in SW480 cells, Reg-R-SW480 cells, Reg-R-SW480-SiRNA- Notch-1 cells, and Reg-R-SW480-SiRNA-control cells. (A) Reg-R-SW480 cells, Reg-R-SW480-SiRNA-Notch-1 cells, and Reg-R-SW480-SiRNA- control cells were treated with regorafenib at concentration 1 mM. Following 24 h incubation, the numbers of viable cells in each well were determined by a proliferation assay as described in “Materials and Methods.” Results were expressed as the percentage of cell growth relative to controls. Each point represents mean SD of three determinations. (B) All cell lines were exposed to regorafenib 1 mM for 24 h, then processed for Western blot analysis of pS473-AKT and AKT. b-actin was used as a control for protein loading. Blots are representative at of at least three separate experiments. (C) Values represent the mean relative ratios SD from three experiments. P < 0.05, P < 0.01 refers to Reg-R-SW480 cells, Reg-R-SW480-SiRNA-Notch-1 cells, and Reg-R-SW480-SiRNA-control cells treated with regorafenib.

Article Snippet: Regorafenib were purchased from Selleck Chemicals (Houston, TX) (Stivarga, 2013) (Fig. 1).

Techniques: Expressing, Control, Concentration Assay, Incubation, Proliferation Assay, Western Blot

Journal: Journal of cellular physiology

Article Title: Involvement of Notch-1 in Resistance to Regorafenib in Colon Cancer Cells.

doi: 10.1002/jcp.25206

Figure Lengend Snippet: Fig. 7. Growth profile and effect of regorafenib on expression p-AKT and AKT in SW480 cells, Reg-R-SW480 cells, SW480-Notch-1 cells treated with regorafenib. (A) SW480 cells, Reg-R-SW480 cells, and SW480-Notch-1 cells were treated with regorafenib. Following 24 h incubation, the numbers of viable cells in each well were determined by a proliferation assay as described in “Materials and Methods.” Results were expressed as the percentage of cell growth relative to controls. Each point represents mean SD of three determinations. (B) SW480 cells, SW480-vector cells, and SW480-Notch-1 cells were exposed to regorafenib 1 mM for 24 h, then processed for Western blot analysis of pS473-AKT and AKT. b-actin was used as a control for protein loading. Blots are representative at of at least three separate experiments. (C) Values represent the mean relative ratios SD from three experiments. P < 0.05, P < 0.01 refers to parental SW480 cells, SW480-Notch-1 cells, and SW480–vector cells were treated with regorafenib 1 mM. (D) SW480 and Reg-R-SW480 cells were incubated with regorafenib 1 mM for 24 h. After 10–14 days incubation, clones (>50 cells) were fixed and manually counted. (E) A representative of four experiments is shown. P < 0.05, P < 0.01 refers to drug treatment group vs. corresponding group without treatment.

Article Snippet: Regorafenib were purchased from Selleck Chemicals (Houston, TX) (Stivarga, 2013) (Fig. 1).

Techniques: Expressing, Incubation, Proliferation Assay, Plasmid Preparation, Western Blot, Control, Clone Assay

Journal: Molecules

Article Title: The Synergistic Anti-Cancer Effects of NVP-BEZ235 and Regorafenib in Hepatocellular Carcinoma

doi: 10.3390/molecules25102454

Figure Lengend Snippet: BEZ235 enhances the anti-proliferation effect of regorafenib in HCC cells. The cell viability analysis for HCC cells treated for 48 h with ( A ) BEZ235 or ( B ) regorafenib. ( C ) Cell viability analysis for the HCC cells treated with various combinations of BEZ235 and regorafenib for 48 h. Scale bar: 50 μm. Data are represented as mean ± S.D. * p < 0.05, and ** p < 0.01 versus untreated control, and BEZ235 or regorafenib alone group.

Article Snippet: BEZ235 was purchased from LC laboratories (Woburn, MA, USA) and regorafenib was obtained from Toronto Research Chemicals (North York, ON, Canada).

Techniques: Control

Journal: Molecules

Article Title: The Synergistic Anti-Cancer Effects of NVP-BEZ235 and Regorafenib in Hepatocellular Carcinoma

doi: 10.3390/molecules25102454

Figure Lengend Snippet: CI and DRI of BEZ235 and Regorafenib combination in HCC.

Article Snippet: BEZ235 was purchased from LC laboratories (Woburn, MA, USA) and regorafenib was obtained from Toronto Research Chemicals (North York, ON, Canada).

Techniques: Concentration Assay

Journal: Molecules

Article Title: The Synergistic Anti-Cancer Effects of NVP-BEZ235 and Regorafenib in Hepatocellular Carcinoma

doi: 10.3390/molecules25102454

Figure Lengend Snippet: The regorafenib-induced apoptosis in HCC cells increases upon treatment with BEZ235. The flow cytometry analysis of cell cycle profiles in HCC cells treated with BEZ235 and regorafenib for 48 h ( A ). The Western blot analysis indicating the expression pattern of cleaved caspase-3 and cleaved PARP ( B ). Data are represented as mean ± S.D. * p < 0.05, and ** p < 0.01 versus untreated control, and BEZ235 or regorafenib alone group.

Article Snippet: BEZ235 was purchased from LC laboratories (Woburn, MA, USA) and regorafenib was obtained from Toronto Research Chemicals (North York, ON, Canada).

Techniques: Flow Cytometry, Western Blot, Expressing, Control

Journal: Molecules

Article Title: The Synergistic Anti-Cancer Effects of NVP-BEZ235 and Regorafenib in Hepatocellular Carcinoma

doi: 10.3390/molecules25102454

Figure Lengend Snippet: Treatment with BEZ235 enhances the anti-migration and -invasion effect of regorafenib in HCC cells. The analysis of transwell migration and invasion assay upon treatment of Hep3B ( A ), HepG2 ( B ) and Huh7 ( C ) cells with BEZ235 and regorafenib for 24 h or 48 h. Data are represented as means ± S.D. * p < 0.05, and ** p < 0.01 versus untreated control, BEZ235 or regorafenib alone group.

Article Snippet: BEZ235 was purchased from LC laboratories (Woburn, MA, USA) and regorafenib was obtained from Toronto Research Chemicals (North York, ON, Canada).

Techniques: Migration, Invasion Assay, Control

Journal: Molecules

Article Title: The Synergistic Anti-Cancer Effects of NVP-BEZ235 and Regorafenib in Hepatocellular Carcinoma

doi: 10.3390/molecules25102454

Figure Lengend Snippet: The combined treatment inhibits the activation of Akt/mTOR signaling pathway in the HCC cells. The panels indicate Western blot analysis for phosphorylation and total protein levels of the members of the Akt/mTOR pathway in the HCC cells treated with BEZ235 and regorafenib for 48 h.

Article Snippet: BEZ235 was purchased from LC laboratories (Woburn, MA, USA) and regorafenib was obtained from Toronto Research Chemicals (North York, ON, Canada).

Techniques: Activation Assay, Western Blot, Phospho-proteomics

Journal: Molecules

Article Title: The Synergistic Anti-Cancer Effects of NVP-BEZ235 and Regorafenib in Hepatocellular Carcinoma

doi: 10.3390/molecules25102454

Figure Lengend Snippet: Model depicting the anti-tumor effect of the combined treatment with BEZ235 and regorafenib in HCC cells. The combined treatment suppressed HCC cell migration and invasion through inhibition of the Akt/mTOR signaling pathway.

Article Snippet: BEZ235 was purchased from LC laboratories (Woburn, MA, USA) and regorafenib was obtained from Toronto Research Chemicals (North York, ON, Canada).

Techniques: Migration, Inhibition

Journal: BMC Complementary Medicine and Therapies

Article Title: Combination of Quercetin or/and siRNA-loaded DDAB-mPEG-PCL hybrid nanoparticles reverse resistance to Regorafenib in colon cancer cells

doi: 10.1186/s12906-022-03787-8

Figure Lengend Snippet: Treatment groups and doses

Article Snippet: Regorafenib (Santa Cruz, CAS: 755037-03-7) and Quercetin (Sigma Aldrich, St. Louis, USA, CAS, 117-39-5) were purchased locally.

Techniques: Control

Journal: BMC Complementary Medicine and Therapies

Article Title: Combination of Quercetin or/and siRNA-loaded DDAB-mPEG-PCL hybrid nanoparticles reverse resistance to Regorafenib in colon cancer cells

doi: 10.1186/s12906-022-03787-8

Figure Lengend Snippet: Cell morphology of Regorafenib-resistant LS-180 cell line at the end of each cyclic is shown. The cells were resistant during four cycles using a fixed dose of 40 µM Regorafenib. As we approached the last cycles, the cells became less sensitive to the drug. To the extent that the behavior of cells over the fourth cycle was no different from that of cells that grew in drug-free media. T: treatment; R: recovery

Article Snippet: Regorafenib (Santa Cruz, CAS: 755037-03-7) and Quercetin (Sigma Aldrich, St. Louis, USA, CAS, 117-39-5) were purchased locally.

Techniques:

Journal: BMC Complementary Medicine and Therapies

Article Title: Combination of Quercetin or/and siRNA-loaded DDAB-mPEG-PCL hybrid nanoparticles reverse resistance to Regorafenib in colon cancer cells

doi: 10.1186/s12906-022-03787-8

Figure Lengend Snippet: A Dose- response survival of LS-180 cells after exposure to increasing concentrations of Regorafenib for 24 h; B Dose- response survival of resistance LS-180 cells after exposure to increasing concentrations of Regorafenib for 24 h; C Dose- response survival of LS-180 cells after exposure to increasing concentrations of Quercetin for 24 h; D Dose- response survival of resistance LS-180 cells after exposure to increasing concentrations of Regorafenib for 24 h

Article Snippet: Regorafenib (Santa Cruz, CAS: 755037-03-7) and Quercetin (Sigma Aldrich, St. Louis, USA, CAS, 117-39-5) were purchased locally.

Techniques:

Journal: BMC Complementary Medicine and Therapies

Article Title: Combination of Quercetin or/and siRNA-loaded DDAB-mPEG-PCL hybrid nanoparticles reverse resistance to Regorafenib in colon cancer cells

doi: 10.1186/s12906-022-03787-8

Figure Lengend Snippet: A The IC 50 values of the Regorafenib group, including free drug (Regorafenib), RDMP, and RDMP/siRNA complex in both resistance and non-resistance cell lines. B The IC 50 values of the Quercetin group, including free drug (Quercetin), QDMP, and QDMP/siRNA complex in both resistance and non-resistance cell lines. RDMP: Reg/DDAB mPEG-PCL nanoparticles; QDMP: Q/DDAB mPEG-PCL nanoparticles; siRNA; small interfering RNA

Article Snippet: Regorafenib (Santa Cruz, CAS: 755037-03-7) and Quercetin (Sigma Aldrich, St. Louis, USA, CAS, 117-39-5) were purchased locally.

Techniques: Small Interfering RNA

Journal: BMC Complementary Medicine and Therapies

Article Title: Combination of Quercetin or/and siRNA-loaded DDAB-mPEG-PCL hybrid nanoparticles reverse resistance to Regorafenib in colon cancer cells

doi: 10.1186/s12906-022-03787-8

Figure Lengend Snippet: The X-axis shows the various concentrations of the diluted drugs (in mg/dL) and the Y-axis shows the degree of absorption reported as Optical density (OD) A Calibration curve in Quercetin ( n = 3) with 95% CI. B Calibration curve Regorafenib ( n = 3) with 95% CI

Article Snippet: Regorafenib (Santa Cruz, CAS: 755037-03-7) and Quercetin (Sigma Aldrich, St. Louis, USA, CAS, 117-39-5) were purchased locally.

Techniques:

Journal: BMC Complementary Medicine and Therapies

Article Title: Combination of Quercetin or/and siRNA-loaded DDAB-mPEG-PCL hybrid nanoparticles reverse resistance to Regorafenib in colon cancer cells

doi: 10.1186/s12906-022-03787-8

Figure Lengend Snippet: A Agarose gel electrophoresis of all real-time PCR products derived from β1-integrin & B-Actin. Left to right in the gel: 50 bp ladder, 28.7µM Quercetin, 65.7 µM Quercetin, 40µM Regorafenib, 40 µM RDMP, 40 µM RDMP and 156nM siRNA, and control cells; B β1 integrin expressions in the non-resistant cells compared with the resistant LS-180 cells

Article Snippet: Regorafenib (Santa Cruz, CAS: 755037-03-7) and Quercetin (Sigma Aldrich, St. Louis, USA, CAS, 117-39-5) were purchased locally.

Techniques: Agarose Gel Electrophoresis, Real-time Polymerase Chain Reaction, Derivative Assay, Control

Journal: BMC Complementary Medicine and Therapies

Article Title: Combination of Quercetin or/and siRNA-loaded DDAB-mPEG-PCL hybrid nanoparticles reverse resistance to Regorafenib in colon cancer cells

doi: 10.1186/s12906-022-03787-8

Figure Lengend Snippet: A Combination therapy using Regorafenib 40µM and increasing level of Quercetin could significantly down-regulate β1 integrin expression in resistant colon cancer cells ( p < 0.01); B There was a significant difference not only between the Regorafenib subgroups but also between the Regorafenib group and untreated state ( p < 0.05); C There was a significant difference between the subgroups of Quercetin and the QDMP/siRNA ( P < 0.01). In the Quercetin group, a decrease in β1 integrin expression in resistant cells was significantly higher in IC 30 than IC 10, IC 30 than IC 20 ( P < 0.01), however, a significant decrease was not observed between IC 20 and IC 10 ( P = 0.09). While, in the QDMP/siRNA group, inter-group comparison of treatment at all doses (IC 30 , IC 20 , and IC 10 ) represented a significant decrease in β1 integrin expression; D No significant relationship was observed in siRNA subgroups. Except, NP/siRNA with untreated state ( p = 0.02).NP: nanoparticle without drug. R: Regorafenib; Q: Quercetin; RDMP: Reg/DDAB mPEG-PCL nanoparticles; QDMP: Q/DDAB mPEG-PCL nanoparticles; siRNA; small interfering RNA

Article Snippet: Regorafenib (Santa Cruz, CAS: 755037-03-7) and Quercetin (Sigma Aldrich, St. Louis, USA, CAS, 117-39-5) were purchased locally.

Techniques: Expressing, Comparison, Small Interfering RNA

Journal: Journal of Biomedical Science

Article Title: Novel FLT3/AURK multikinase inhibitor is efficacious against sorafenib-refractory and sorafenib-resistant hepatocellular carcinoma

doi: 10.1186/s12929-022-00788-0

Figure Lengend Snippet: Effect of DBPR114 and regorafenib on sorafenib-refractory and sorafenib-acquired resistant human HCC xenograft tumors. A and B Tumor growth curves and body weight changes from baseline (%) for the control and treated HA22T/VGH xenograft tumors. C Tumor growth curves for the control and treated sorafenib-acquired resistant Huh7 xenograft tumors. D Survival in the control and treated sorafenib-acquired resistant Huh7 xenograft tumor groups. DBPR114 (40 mg/kg) was administered once a week intravenously for 6 weeks for the HA22T/VGH xenograft tumors and 3 weeks for the sorafenib-acquired resistant Huh7 xenograft tumors. Sorafenib and regorafenib were administered at 30 mg/kg once a day, 5 days per week by oral gavage for 40 days for the HA22T/VGH xenograft tumors and 25 days for the Huh7 xenograft tumors. Mean ± SEM, n = 8 mice per group for both xenograft tumors. * p < 0.05 vs. vehicle control measured using one-way ANOVA and Bonferroni posttest comparison. & p < 0.01 vs. control, $ p < 0.05 vs. regorafenib, measured using Mantel–Cox test

Article Snippet: Sorafenib and regorafenib were purchased from BOC Sciences (Shirley, NY, USA), and VX680 and nocodazole from Abcam (Cambridge, MA, USA).

Techniques: Control, Comparison

Journal: International Journal of Molecular Sciences

Article Title: Chlorogenic Acid Improves the Regorafenib Effects in Human Hepatocellular Carcinoma Cells

doi: 10.3390/ijms19051518

Figure Lengend Snippet: CGA potentiates the inhibitory effect of Regorafenib on cell proliferation. PLC/PRF5 and HepG2 cells were cultured with increasing concentrations of Regorafenib and CGA alone or in combination. MTT assay was assessed after 48 h. For each experimental condition, the results of three independent experiments were quantified and the values expressed as mean ± SD, and plotted in the relative graph. * p < 0.05; ** p < 0.001; *** p < 0.0001.

Article Snippet: Regorafenib was gifted from Bayer Corp (West Haven, CT, USA); Chlorogenic acid (CGA) was purchased from Sigma-Aldrich (Milan, Italy); PLC/PRF/5 and HepG2 human HCC cell lines were purchased from the National Institute of Biomedical Innovation JCRB Cell Bank (Osaka, Japan).

Techniques: Cell Culture, MTT Assay

Journal: International Journal of Molecular Sciences

Article Title: Chlorogenic Acid Improves the Regorafenib Effects in Human Hepatocellular Carcinoma Cells

doi: 10.3390/ijms19051518

Figure Lengend Snippet: Combination index (CI) values calculated for each combined drug treatments in PLC/PRF/5 and HepG2 cells. Each value was derived from the method described by Chou and Talalay and implemented in CompuSyn software. R = Regorafenib; CGA = Chlorogenic Acid.

Article Snippet: Regorafenib was gifted from Bayer Corp (West Haven, CT, USA); Chlorogenic acid (CGA) was purchased from Sigma-Aldrich (Milan, Italy); PLC/PRF/5 and HepG2 human HCC cell lines were purchased from the National Institute of Biomedical Innovation JCRB Cell Bank (Osaka, Japan).

Techniques: Derivative Assay, Software

Journal: International Journal of Molecular Sciences

Article Title: Chlorogenic Acid Improves the Regorafenib Effects in Human Hepatocellular Carcinoma Cells

doi: 10.3390/ijms19051518

Figure Lengend Snippet: CGA potentiates the Regorafenib-mediated growth inhibition by modifying Ki67 expression. Ki67 staining in PLC/PRF5 and HepG2 cells cultured with 1 µM (PLC/PRF/5) or 0.1 μM (HepG2) Regorafenib and 100 µM CGA alone or in combination. For each experimental condition, the results of three independent experiments were quantified and the intensity fluorescent values, expressed as mean ± SD, and plotted in the relative graph. * p < 0.05; ** p < 0.001; *** p < 0.0001. Scale bar: 100 μm.

Article Snippet: Regorafenib was gifted from Bayer Corp (West Haven, CT, USA); Chlorogenic acid (CGA) was purchased from Sigma-Aldrich (Milan, Italy); PLC/PRF/5 and HepG2 human HCC cell lines were purchased from the National Institute of Biomedical Innovation JCRB Cell Bank (Osaka, Japan).

Techniques: Inhibition, Expressing, Staining, Cell Culture

Journal: International Journal of Molecular Sciences

Article Title: Chlorogenic Acid Improves the Regorafenib Effects in Human Hepatocellular Carcinoma Cells

doi: 10.3390/ijms19051518

Figure Lengend Snippet: CGA potentiates the Regorafenib-mediated growth inhibition by modifying cell cycle progression. PLC/PRF5 and HepG2 cells cultured with 1 µM (PLC/PRF/5) or 0.1 µM (HepG2) Regorafenib and 100 µM CGA alone or in combination, were synchronized in the S phase of the cell cycle using thymidine (0.2 M) (T0). After 3 h from blockrelease (T1), the cells were processed with the Cell Cycle Kit and analyzed with Muse Cell Analyzer to evaluate the percentage of cells in G0/G1, S and G2/M phases. An example of cell cycle progression in different treatment conditions are shown in the panels. The results of three independent experiments expressed as mean ± SD, are plotted in the relative graphs. * p < 0.05; ** p < 0.001*** p < 0.0001.

Article Snippet: Regorafenib was gifted from Bayer Corp (West Haven, CT, USA); Chlorogenic acid (CGA) was purchased from Sigma-Aldrich (Milan, Italy); PLC/PRF/5 and HepG2 human HCC cell lines were purchased from the National Institute of Biomedical Innovation JCRB Cell Bank (Osaka, Japan).

Techniques: Inhibition, Cell Culture

Journal: International Journal of Molecular Sciences

Article Title: Chlorogenic Acid Improves the Regorafenib Effects in Human Hepatocellular Carcinoma Cells

doi: 10.3390/ijms19051518

Figure Lengend Snippet: CGA potentiates the pro-apoptotic effects of Regorafenib. PLC/PRF5 and HepG2cells were cultured with 1 µM (PLC/PRF/5) or 0.1 µM (HepG2) Regorafenib and 100 µM CGA alone or in combination, were analyzed for the percentage of live, early/ late apoptotic and dead cells. Muse Annexin V ( a ), Muse Caspase-3/7 ( b ) and Bcl-2 activation ( c ). Cell Assays were performed after 48 h. The results of three independent experiments are expressed as means ± SD. * p < 0.05; ** p < 0.001; *** p < 0.0001. ( d ) Western blot showing the expression levels of some proteins involved in apoptosis process after 48 h of single or combined treatments.

Article Snippet: Regorafenib was gifted from Bayer Corp (West Haven, CT, USA); Chlorogenic acid (CGA) was purchased from Sigma-Aldrich (Milan, Italy); PLC/PRF/5 and HepG2 human HCC cell lines were purchased from the National Institute of Biomedical Innovation JCRB Cell Bank (Osaka, Japan).

Techniques: Cell Culture, Activation Assay, Western Blot, Expressing

Journal: International Journal of Molecular Sciences

Article Title: Chlorogenic Acid Improves the Regorafenib Effects in Human Hepatocellular Carcinoma Cells

doi: 10.3390/ijms19051518

Figure Lengend Snippet: CGA potentiates the Regorafenib modulation of MAPK and PI3K/Akt pathways. PLC/PRF5 and HepG2 cells were cultured with 1 µM (PLC/PRF/5) or 0.1 µM (HepG2) Regorafenib and 100 µM CGA alone or in combination. ( a ) Western blot showing the expression levels of some proteins involved in MAPK and PI3K/Akt pathways after 48 h of single or combined treatments. ( b ) The Muse MAPK Activation Kit was used to evaluate ERK phosphorylation relative to total ERK expression after 3 h, and the Muse PI3K Activation dual detection Kit was used to detect Akt phosphorylation (Ser473) relative to total Akt expression after 48 h. The results of three independent experiments, expressed as mean ± SD, are plotted in the relative graph. * p < 0.05; ** p < 0.001; *** p < 0.0001. ( c ) Western blot showing the expression levels of P-ERK after 15 min and P-Akt after 48 h of single or combined treatments.

Article Snippet: Regorafenib was gifted from Bayer Corp (West Haven, CT, USA); Chlorogenic acid (CGA) was purchased from Sigma-Aldrich (Milan, Italy); PLC/PRF/5 and HepG2 human HCC cell lines were purchased from the National Institute of Biomedical Innovation JCRB Cell Bank (Osaka, Japan).

Techniques: Cell Culture, Western Blot, Expressing, Activation Assay, Phospho-proteomics

Journal: International Journal of Molecular Sciences

Article Title: Chlorogenic Acid Improves the Regorafenib Effects in Human Hepatocellular Carcinoma Cells

doi: 10.3390/ijms19051518

Figure Lengend Snippet: CGA potentiates the Regorafenib-mediated inhibition of migration. PLC/PRF5 and HepG2 cells were cultured with 1 µM (PLC/PRF/5) or 0.1 µM (HepG2) Regorafenib and 100 µM CGA alone or in combination. The migration assay was performed in cells seeded on collagen I coated wells and treated as described. Representative photographs (T0 and T2) were shown ( upper panel 16× magnification). The relative values were expressed as percentage of migration, where 100% represents the detection zone completely closed. The results of three independent experiments, expressed as mean ± SD, are plotted in the relative graph ( lower panel ). * p < 0.05; ** p < 0.001; *** p < 0.0001.

Article Snippet: Regorafenib was gifted from Bayer Corp (West Haven, CT, USA); Chlorogenic acid (CGA) was purchased from Sigma-Aldrich (Milan, Italy); PLC/PRF/5 and HepG2 human HCC cell lines were purchased from the National Institute of Biomedical Innovation JCRB Cell Bank (Osaka, Japan).

Techniques: Inhibition, Migration, Cell Culture