Journal: Oncology Research

Article Title: Glutamic Acid–Chelated Cobalt Stabilizes G-Quadruplexes and Selectively Suppresses Hepatocellular Carcinoma Growth

doi: 10.32604/or.2026.074144

Figure Lengend Snippet: (AI-PRS optimization of combination therapies: GACC, metformin, and regorafenib. ( A–C ) Viability responses for the tested 3 × 3 × 3 dose grid of GACC (9.54, 28.61, 85.85 µM), metformin (0.85, 2.55, and 7.65 mM), and regorafenib (0.5, 1.5, 4.5 µM) in hepatoma cells (PLC5) and non-transformed hepatocytes (THLE-2), as indicated. ( D ) AI-phenotypic response surface (AI-PRS) fit across the dose grid used to identify dose regions with improved efficacy–toxicity balance. ( E ) Observed vs. AI-PRS–predicted viability for the off-grid optimal dose combination in PLC/PRF/5 and THLE-2. Data are presented as mean ± SEM; n values and statistical methods are provided in . Statistical annotations: ns, not significant; * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001.)

Article Snippet: Metformin hydrochloride (MedChemExpress, HY-17471A/CS-1851; CAS 1115-70-4; MedChemExpress, Monmouth Junction, NJ, USA), regorafenib (TargetMol, T1792; CAS 755037-03-7, TargetMol, Boston, MA, USA), and pyridostatin (RR82) trifluoroacetate salt (PDS, Selleck Chemicals, S7444; Houston, TX, USA) were dissolved in dimethyl sulfoxide (DMSO, Sigma-Aldrich, D4540; St. Louis, MO, USA) to prepare stock solutions and diluted into assay media immediately before use.

Techniques: Transformation Assay

Journal: Oncology Research

Article Title: Glutamic Acid–Chelated Cobalt Stabilizes G-Quadruplexes and Selectively Suppresses Hepatocellular Carcinoma Growth

doi: 10.32604/or.2026.074144

Figure Lengend Snippet: Combination therapies of GACC, metformin, and regorafenib for Hepatocellular Carcinoma (HCC) treatment in tert transgenic zebrafish larvae and HBx, src, p53-, RPIA (HSPR) adult zebrafish model. ( A ) Evaluation of drug-induced side effects by measuring body length in tert transgenic zebrafish larvae before and after treatment, assessing potential toxicity and systemic impact. ( B – D ) qPCR analysis of cell proliferation markers ( ccne1 , cdk1 , cdk2 ) in tert transgenic zebrafish larvae, demonstrating significant downregulation following combination therapy. ( E , F ) Histological examination of liver cancer hallmarks in HSPR transgenic zebrafish using hematoxylin and eosin (H&E) staining, revealing a significant reduction in mitotic figures, karyomegalic cells, and hepatocellular dysplasia following combination therapy. n = 10 embryos/group; one-way ANOVA with Dunnett’s post-hoc vs. vehicle; p thresholds as shown. * p < 0.05, ** p < 0.01, *** p < 0.005, **** p < 0.001, ns, not significant.

Article Snippet: Metformin hydrochloride (MedChemExpress, HY-17471A/CS-1851; CAS 1115-70-4; MedChemExpress, Monmouth Junction, NJ, USA), regorafenib (TargetMol, T1792; CAS 755037-03-7, TargetMol, Boston, MA, USA), and pyridostatin (RR82) trifluoroacetate salt (PDS, Selleck Chemicals, S7444; Houston, TX, USA) were dissolved in dimethyl sulfoxide (DMSO, Sigma-Aldrich, D4540; St. Louis, MO, USA) to prepare stock solutions and diluted into assay media immediately before use.

Techniques: Transgenic Assay, Staining

Journal: Medeniyet Medical Journal

Article Title: Overcoming Regorafenib Resistance in Colorectal Cancer: Synergistic Efficacy of Cilengitide in HT29 and WiDr Cells

doi: 10.4274/MMJ.galenos.2026.65848

Figure Lengend Snippet: Dose-response curves of P and R colorectal cancer cells treated with regorafenib. Cell viability was measured after exposure to increasing concentrations of regorafenib (0.001-100 µM) in HT29 (A) and WiDr (B) cells. Data are presented as mean ± SD. IC 50 values were calculated for each cell line and are indicated by dashed vertical lines. C) Bar chart of IC 50 values. One-way ANOVA followed by Tukey’s post-hoc test was used for the analyses. Parental (P) and resistant (R) SD: Standard deviation

Article Snippet: Regorafenib (Selleckchem, Cat. No. S1178) and cilengitide (Selleckchem, Cat. No. S6387) were prepared as 100 mM stock solutions in DMSO.5×10 3 cells per well were seeded in 96-well plates and treated with increasing concentrations of regorafenib or cilengitide (0.001-100 μM) for 72 h. MTT reagent (0.5 mg/mL, Sigma-Aldrich) was added, and the samples were incubated for 4 h. Formazan crystals were solubilized with DMSO, and absorbance was measured at 570 nm using a microplate reader (BioTek, USA).

Techniques: Standard Deviation

Journal: Medeniyet Medical Journal

Article Title: Overcoming Regorafenib Resistance in Colorectal Cancer: Synergistic Efficacy of Cilengitide in HT29 and WiDr Cells

doi: 10.4274/MMJ.galenos.2026.65848

Figure Lengend Snippet: Clonogenic survival assay of HT29 and WiDr colon cancer cells. A) Images of clonogenic assays showing the effects of DMSO (vehicle control), regorafenib (IC 50 ), cilengitide (IC 50 ), and the combination of both drugs on parental (P) and regorafenib-resistant (R) HT29 and WiDr cell lines. Colony formation, visualized by crystal violet staining, indicates long-term cell survival and proliferation. B) Corresponding bar graphs quantifying the surviving fraction of HT29-P vs. HT29-R (top) and WiDr-P vs. WiDr-R (bottom) cells after 14 days of treatment. C) p-values data are presented as mean surviving fraction ± standard deviation from three independent experiments. One-way ANOVA followed by Tukey’s post-hoc test was used for the analyses. *p<0.05 DMSO: Dimethyl sulfoxide

Article Snippet: Regorafenib (Selleckchem, Cat. No. S1178) and cilengitide (Selleckchem, Cat. No. S6387) were prepared as 100 mM stock solutions in DMSO.5×10 3 cells per well were seeded in 96-well plates and treated with increasing concentrations of regorafenib or cilengitide (0.001-100 μM) for 72 h. MTT reagent (0.5 mg/mL, Sigma-Aldrich) was added, and the samples were incubated for 4 h. Formazan crystals were solubilized with DMSO, and absorbance was measured at 570 nm using a microplate reader (BioTek, USA).

Techniques: Clonogenic Cell Survival Assay, Control, Staining, Standard Deviation

Journal: Medeniyet Medical Journal

Article Title: Overcoming Regorafenib Resistance in Colorectal Cancer: Synergistic Efficacy of Cilengitide in HT29 and WiDr Cells

doi: 10.4274/MMJ.galenos.2026.65848

Figure Lengend Snippet: Synergistic analysis of the regorafenib and cilengitide combination. The figure presents the Chou-Talalay analysis of the drug combination’s effects on colon cancer cell lines. The CI (Combination Index)-Fa (fraction affected) plots show the CI values across various levels of drug effect. A CI<1 indicates synergy, CI=1 is an additive effect, and CI>1 is antagonism. The isobolograms below each plot visually represent this synergy, with points falling below the line of additivity. The top row shows the synergy analysis for the HT29-P (parental) and HT29-R (regorafenib-resistant) cell lines, while the bottom row shows the analysis for the WiDr-P and WiDr-R cell lines. The data consistently demonstrates a synergistic interaction for the combination therapy across all cell lines.

Article Snippet: Regorafenib (Selleckchem, Cat. No. S1178) and cilengitide (Selleckchem, Cat. No. S6387) were prepared as 100 mM stock solutions in DMSO.5×10 3 cells per well were seeded in 96-well plates and treated with increasing concentrations of regorafenib or cilengitide (0.001-100 μM) for 72 h. MTT reagent (0.5 mg/mL, Sigma-Aldrich) was added, and the samples were incubated for 4 h. Formazan crystals were solubilized with DMSO, and absorbance was measured at 570 nm using a microplate reader (BioTek, USA).

Techniques:

Journal: Oncology Reports

Article Title: Senescence-associated IL-33 secretion undermines sorafenib efficacy in hepatocellular carcinoma via immune evasion

doi: 10.3892/or.2025.8987

Figure Lengend Snippet: Sorafenib and regorafenib induce IL-33 upregulation and secretion in hepatocellular carcinoma cells. (A) Huh-7 cells were treated with sorafenib or regorafenib at the indicated concentrations. Cell death was assessed using PI staining through flow cytometry. Representative flow cytometry histograms of PI staining in Huh7 cells under the indicated treatment conditions are shown (left). Quantification of PI-positive cells (%) is presented (right). (B and C) Huh-7 cells were treated with DMEM, sorafenib (10 µM), or regorafenib (20 µM) for the indicated time. (B) The protein expression of IL-33 was analyzed by western blotting. (C) The mRNA level of IL33 was determined by RT-qPCR. (D and E) Huh-7 cells were treated with rIL-33 at the indicated concentrations for 48 h. (D, upper) The mRNA level of IL33 was determined by RT-qPCR. (D, lower) The cell surface expression of ST2L was measured by flow cytometry. (E) The protein expression of IL-33 was analyzed by western blotting. (F) Huh-7 cells were treated with sorafenib (left) and regorafenib (right) with or without α-IL-33 antibodies (10 µg/ml). The mRNA level of IL33 was determined by RT-qPCR. *P<0.05, **P<0.01 and ***P<0.001. PI, propidium iodide; RT-qPCR, reverse transcription-quantitative PCR; Sup, supernatant; WCL, whole cell lysate.

Article Snippet: Huh-7 cells (5×10 3 cells/well) were seeded in 96-well plates and treated with sorafenib (cat. no. SC-220125; Santa Cruz Biotechnology) or regorafenib (cat. no. SC-477163; Santa Cruz Biotechnology) at the indicated concentrations for 24 to 96 h. Cell death was assessed using the FITC Annexin V Apoptosis Detection Kit with propidium iodide (cat. no. 640914; BioLegend, Inc.) according to the manufacturer's instructions.

Techniques: Staining, Flow Cytometry, Expressing, Western Blot, Quantitative RT-PCR, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Oncology Reports

Article Title: Senescence-associated IL-33 secretion undermines sorafenib efficacy in hepatocellular carcinoma via immune evasion

doi: 10.3892/or.2025.8987

Figure Lengend Snippet: Sorafenib and regorafenib trigger cellular senescence in hepatocellular carcinoma cells. (A) Huh-7 cells were treated with sorafenib (1 and 10 µM, upper) and regorafenib (10 and 20 µM, lower). Cell numbers were calculated using a hemocytometer at the indicated time points. (B) Huh-7 cells were treated with sorafenib (10 µM) and regorafenib (20 µM) for 96 h. The SA-β-gal activity was detected using a senescence detection kit. (C) Huh-7 cells were treated with sorafenib (10 µM, left) and regorafenib (20 µM, right) for the indicated time. The protein expression of p16 and p21 was analyzed by western blotting.

Article Snippet: Huh-7 cells (5×10 3 cells/well) were seeded in 96-well plates and treated with sorafenib (cat. no. SC-220125; Santa Cruz Biotechnology) or regorafenib (cat. no. SC-477163; Santa Cruz Biotechnology) at the indicated concentrations for 24 to 96 h. Cell death was assessed using the FITC Annexin V Apoptosis Detection Kit with propidium iodide (cat. no. 640914; BioLegend, Inc.) according to the manufacturer's instructions.

Techniques: Activity Assay, Expressing, Western Blot

Journal: Oncology Reports

Article Title: Senescence-associated IL-33 secretion undermines sorafenib efficacy in hepatocellular carcinoma via immune evasion

doi: 10.3892/or.2025.8987

Figure Lengend Snippet: Sorafenib and regorafenib enhance PD-L1 expression via IL-33 signaling in hepatocellular carcinoma cells. (A) Huh-7 cells were treated with sorafenib (10 µM) and regorafenib (20 µM) for 96 h. The cell surface expression of PD-L1 was measured by flow cytometry. (B) Huh-7 cells were treated with sorafenib (10 µM, upper) and regorafenib (20 µM, lower) with or without treatment of α-IL-33 antibodies (10 µg/ml) for 96 h. The cell surface expression of PD-L1 was measured by flow cytometry. (C and D) Huh-7 cells were treated with rIL-33 at the indicated concentrations for 96 h. (C) The mRNA level of CD274 was determined by reverse transcription-quantitative PCR. (D) Huh-7 cells were treated with rIL-33 (100 ng/ml) with or without treatment of JSH-23 (50 µM). The cell surface expression of PD-L1 was measured by flow cytometry. *P<0.05, **P<0.01 and ***P<0.001. PD-L1, programmed death ligand 1.

Article Snippet: Huh-7 cells (5×10 3 cells/well) were seeded in 96-well plates and treated with sorafenib (cat. no. SC-220125; Santa Cruz Biotechnology) or regorafenib (cat. no. SC-477163; Santa Cruz Biotechnology) at the indicated concentrations for 24 to 96 h. Cell death was assessed using the FITC Annexin V Apoptosis Detection Kit with propidium iodide (cat. no. 640914; BioLegend, Inc.) according to the manufacturer's instructions.

Techniques: Expressing, Flow Cytometry, Reverse Transcription, Real-time Polymerase Chain Reaction

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