Journal: Translational Gastroenterology and Hepatology

Article Title: Fedratinib treatment correlates with suppressed STAT3/AKT/calpain-1 signaling and inhibits hepatocellular carcinoma progression

doi: 10.21037/tgh-2026-0115

Figure Lengend Snippet: Fedratinib inhibits HCC cell viability. (A) Viability of Hepa1-6 cells treated with increasing concentrations of fedratinib for 24 h (B) Dose-response curve and IC50 determination in Hepa1-6 cells. (C) Viability of Huh7 cells treated with increasing concentrations of fedratinib for 24 h. (D) Dose-response curve and IC 50 determination in Huh7 cells. All data are expressed as mean ± SD (n=3 per group), individual data points from independent biological replicates (n=3) are overlaid on the bars. HCC, hepatocellular carcinoma; IC 50 , half-maximal inhibitory concentration; SD, standard deviation.

Article Snippet: Fedratinib hydrochloride hydrate powder was purchased from TargetMol Chemicals Inc. (Boston, MA, USA).

Techniques: Concentration Assay, Standard Deviation

Journal: Translational Gastroenterology and Hepatology

Article Title: Fedratinib treatment correlates with suppressed STAT3/AKT/calpain-1 signaling and inhibits hepatocellular carcinoma progression

doi: 10.21037/tgh-2026-0115

Figure Lengend Snippet: Fedratinib inhibits HCC cell migration. (A) Representative wound healing images of Hepa1-6 cells treated with fedratinib (0, 5, and 10 μM) at 0, 24, and 48 h. (B) Quantification of migration rates in Hepa1-6 cells. (C) Representative wound healing images of Huh7 cells treated with fedratinib (0, 1, and 2 μM) at 0, 24, and 48 h. (D) Quantification of migration rates in Huh7 cells. All data are expressed as mean ± SD (n=3 per group), individual data points from independent biological replicates (n=3) are overlaid on the bars. HCC, hepatocellular carcinoma; SD, standard deviation.

Article Snippet: Fedratinib hydrochloride hydrate powder was purchased from TargetMol Chemicals Inc. (Boston, MA, USA).

Techniques: Migration, Standard Deviation

Journal: Translational Gastroenterology and Hepatology

Article Title: Fedratinib treatment correlates with suppressed STAT3/AKT/calpain-1 signaling and inhibits hepatocellular carcinoma progression

doi: 10.21037/tgh-2026-0115

Figure Lengend Snippet: Fedratinib inhibits migration and invasion of HCC cells. (A) Representative crystal violet-stained images of Transwell migration assay in Hepa1-6 cells treated with fedratinib (0, 5 and 10 μM). (B) Quantitative analysis of migrated Hepa1-6 cells. (C) Representative crystal violet-stained images of Transwell invasion assay in Hepa1-6 cells treated with fedratinib (0, 5 and 10 μM). (D) Quantitative analysis of invaded Hepa1-6 cells. (E) Representative crystal violet-stained images of Transwell migration assay in Huh7 cells treated with fedratinib (0, 1 and 2 μM). (F) Quantitative analysis of migrated Huh7 cells. (G) Representative crystal violet-stained images of Transwell invasion assay in Huh7 cells treated with fedratinib (0, 1 and 2 μM). (H) Quantitative analysis of invaded Huh7 cells. All data are expressed as mean ± SD (n=3 per group), individual data points from independent biological replicates (n=3) are overlaid on the bars. ****, P<0.0001; ***, P<0.001; **, P<0.01; *, P<0.05. HCC, hepatocellular carcinoma; SD, standard deviation.

Article Snippet: Fedratinib hydrochloride hydrate powder was purchased from TargetMol Chemicals Inc. (Boston, MA, USA).

Techniques: Migration, Staining, Transwell Migration Assay, Analysis, Transwell Invasion Assay, Standard Deviation

Journal: Translational Gastroenterology and Hepatology

Article Title: Fedratinib treatment correlates with suppressed STAT3/AKT/calpain-1 signaling and inhibits hepatocellular carcinoma progression

doi: 10.21037/tgh-2026-0115

Figure Lengend Snippet: Fedratinib induces apoptosis in HCC cells. (A) Representative flow cytometry plots of Annexin V/PI staining in Hepa1-6 cells treated with fedratinib (0, 5 and 10 μM). (B) Quantitative analysis of apoptotic Hepa1-6 cells. (C) Representative flow cytometry plots in Huh7 cells treated with fedratinib (0, 1 and 2 μM). (D) Quantification of apoptotic Huh7 cells. All data are presented as mean ± SD (n=3 per group), individual data points from independent biological replicates (n=3) are overlaid on the bars. ****, P<0.0001. HCC, hepatocellular carcinoma; SD, standard deviation.

Article Snippet: Fedratinib hydrochloride hydrate powder was purchased from TargetMol Chemicals Inc. (Boston, MA, USA).

Techniques: Flow Cytometry, Staining, Analysis, Standard Deviation

Journal: Translational Gastroenterology and Hepatology

Article Title: Fedratinib treatment correlates with suppressed STAT3/AKT/calpain-1 signaling and inhibits hepatocellular carcinoma progression

doi: 10.21037/tgh-2026-0115

Figure Lengend Snippet: Fedratinib induces cell cycle arrest in HCC cells. (A) Representative flow cytometry histograms showing cell cycle distribution in Hepa1-6 cells treated with fedratinib (0, 5, and 10 μM). (B) Quantitative analysis of cell cycle phase distribution in Hepa1-6 cells. (C) Representative flow cytometry histograms in Huh7 cells treated with fedratinib (0, 1, and 2 μM). (D) Quantitative analysis of cell cycle phase distribution in Huh7 cells. All data are expressed as mean ± SD (n=3 per group). HCC, hepatocellular carcinoma; SD, standard deviation.

Article Snippet: Fedratinib hydrochloride hydrate powder was purchased from TargetMol Chemicals Inc. (Boston, MA, USA).

Techniques: Flow Cytometry, Analysis, Standard Deviation

Journal: Translational Gastroenterology and Hepatology

Article Title: Fedratinib treatment correlates with suppressed STAT3/AKT/calpain-1 signaling and inhibits hepatocellular carcinoma progression

doi: 10.21037/tgh-2026-0115

Figure Lengend Snippet: Fedratinib generates bulk association signals with intact Hepa1-6 cells in whole-cell SPR analysis. (A) Representative bright-field and SPR images showing cellular ROIs used for data extraction. (B) Distribution of kinetic parameters derived from global fitting. (C) Summary of apparent kinetic constants (KD, ka, kd) obtained from the 1:1 incomplete dissociation model. (D) Sensorgrams showing concentration-dependent association responses; the x-axis represents time in seconds (s). (E) Equilibrium binding analysis demonstrating the correlation between fedratinib concentration and association response intensity. CI, confidence interval; ka, association rate constant; kd, dissociation rate constant; KD, equilibrium dissociation constant; RF, range factor; ROIs, regions of interest; SPR, surface plasmon resonance.

Article Snippet: Fedratinib hydrochloride hydrate powder was purchased from TargetMol Chemicals Inc. (Boston, MA, USA).

Techniques: Analysis, Extraction, Derivative Assay, Global Fitting, Concentration Assay, Binding Assay, Association Rate Constant, Dissociation Rate Constant, Affinity Constant, SPR Assay

Journal: Translational Gastroenterology and Hepatology

Article Title: Fedratinib treatment correlates with suppressed STAT3/AKT/calpain-1 signaling and inhibits hepatocellular carcinoma progression

doi: 10.21037/tgh-2026-0115

Figure Lengend Snippet: Fedratinib generates bulk association signals with intact Huh7 cells in whole-cell SPR analysis. (A) Representative bright-field and SPR images showing cellular ROIs used for data extraction. (B) Distribution of kinetic parameters derived from global fitting. (C) Summary of apparent kinetic constants (KD, ka, kd) obtained from the 1:1 incomplete dissociation model. (D) Sensorgrams showing concentration-dependent association responses; the x-axis represents time in seconds (s). (E) Equilibrium binding analysis demonstrating the correlation between fedratinib concentration and association response intensity. CI, confidence interval; ka, association rate constant; kd, dissociation rate constant; KD, equilibrium dissociation constant; RF, range factor; ROIs, regions of interest; SPR, surface plasmon resonance.

Article Snippet: Fedratinib hydrochloride hydrate powder was purchased from TargetMol Chemicals Inc. (Boston, MA, USA).

Techniques: Analysis, Extraction, Derivative Assay, Global Fitting, Concentration Assay, Binding Assay, Association Rate Constant, Dissociation Rate Constant, Affinity Constant, SPR Assay

Journal: Translational Gastroenterology and Hepatology

Article Title: Fedratinib treatment correlates with suppressed STAT3/AKT/calpain-1 signaling and inhibits hepatocellular carcinoma progression

doi: 10.21037/tgh-2026-0115

Figure Lengend Snippet: Fedratinib inhibits Hepa1-6-Luc allograft tumor growth in vivo. (A-B) Representative photographs of excised tumors from control and fedratinib-treated mice at the end of the experiment. (C) Quantitative analysis of tumor volume at endpoint. (D) Quantitative analysis of tumor weight at endpoint. (E) Tumor growth curves over 14 days of treatment (n=6 per group). (F) Representative in vivo bioluminescence images of tumors from control and fedratinib-treated mice at day 0, 7 and 14. (G) Quantification of bioluminescence intensity over time. (H) Body weight changes of mice during the treatment period. All data are presented as mean ± SD (n=6 per group), individual data points from independent biological replicates (n=6) are overlaid on the bars. ns, no significance; ****, P<0.0001; **, P<0.01; *, P<0.05. SD, standard deviation.

Article Snippet: Fedratinib hydrochloride hydrate powder was purchased from TargetMol Chemicals Inc. (Boston, MA, USA).

Techniques: In Vivo, Control, Analysis, Standard Deviation

Journal: Translational Gastroenterology and Hepatology

Article Title: Fedratinib treatment correlates with suppressed STAT3/AKT/calpain-1 signaling and inhibits hepatocellular carcinoma progression

doi: 10.21037/tgh-2026-0115

Figure Lengend Snippet: Fedratinib inhibits STAT3/AKT phosphorylation and downregulates Bcl-2 expression in Hepa1-6 cells and allograft tumors. (A) Representative western blot images showing the expression of STAT3, p-STAT3, AKT, p-AKT, and Bcl-2 in Hepa1-6 cells treated with Fedratinib (0, 5 and 10 μM) for 24 h. β-actin was used as a loading control. Molecular weights: STAT3 88 kDa, p-STAT3 86 kDa, AKT 56 kDa, p-AKT 56 kDa, β-actin 42 kDa, Bcl-2 26 kDa. (B) Densitometric quantification of protein expression levels from three independent experiments (n=3). Data are normalized to β-actin and expressed as relative to the control group. (C) Representative western blot images of STAT3, p-STAT3, AKT, p-AKT, and Bcl-2 in tumor tissue from control and Fedratinib-treated mice bearing Hepa1-6-Luc allograft. β-actin was used as a loading control. Molecular weights: STAT3 88 kDa, p-STAT3 86 kDa, AKT 56 kDa, p-AKT 56 kDa, β-actin 42 kDa, Bcl-2 26 kDa. (D) Quantitative analysis of protein expression levels from three independent experiments (n=3). All data are presented as mean ± SD (n=3 per group), individual data points from independent biological replicates (n=3) are overlaid on the bars. ns, no significance; ****, P<0.0001; **, P<0.01. SD, standard deviation.

Article Snippet: Fedratinib hydrochloride hydrate powder was purchased from TargetMol Chemicals Inc. (Boston, MA, USA).

Techniques: Phospho-proteomics, Expressing, Western Blot, Control, Tissue, Analysis, Standard Deviation

Journal: Translational Gastroenterology and Hepatology

Article Title: Fedratinib treatment correlates with suppressed STAT3/AKT/calpain-1 signaling and inhibits hepatocellular carcinoma progression

doi: 10.21037/tgh-2026-0115

Figure Lengend Snippet: Fedratinib inhibits calpain-1 expression and activity in Hepa1-6 cells and allograft tumors. (A) Representative western blot images showing calpain-1 protein expression in Hepa1-6 cells treated with Fedratinib (0, 5, 10 μM) for 24 h. GAPDH was used as a loading control. Molecular weights: calpain-1 80 kDa, GAPDH 36 kDa. (B) Densitometric quantification of calpain-1 protein levels three independent experiments. Data are normalized to GAPDH and expressed as relative to the control group. (C) Calpain-1 enzymatic activity in Hepa1-6 cells treated with Fedratinib (0, 5, 10 μM) for 24 h, measured by fluorometric assay. (D) Representative western blot images of calpain-1 expression in tumor tissues from control and Fedratinib-treated mice bearing Hepa1-6-Luc allograft. (E) Quantitative analysis of calpain-1 protein levels three independent experiments. (F) Calpain-1 enzymatic activity in tumor tissues from control and fedratinib treated mice. All data are presented as mean ± SD. For in vitro experiments, n=3 per group; for in vivo experiments, n=6 per group. Individual data points from independent biological replicates are overlaid on the bars. ****, P<0.0001; ***, P<0.001. SD, standard deviation.

Article Snippet: Fedratinib hydrochloride hydrate powder was purchased from TargetMol Chemicals Inc. (Boston, MA, USA).

Techniques: Expressing, Activity Assay, Western Blot, Control, Analysis, In Vitro, In Vivo, Standard Deviation

Journal: Translational Gastroenterology and Hepatology

Article Title: Fedratinib treatment correlates with suppressed STAT3/AKT/calpain-1 signaling and inhibits hepatocellular carcinoma progression

doi: 10.21037/tgh-2026-0115

Figure Lengend Snippet: Serum biochemical evaluation in Hepa1-6-Luc tumor-bearing mice. Serum biochemical parameters were assessed in mice bearing Hepa1-6-Luc cell-derived tumors treated with vehicle (control) or fedratinib. Liver function-related markers (ALB, ALP, ALT and AST), renal function-related markers (CRE, UREA, and UA), and additional indices (HBDH, CHE, CK, GLB, and LDH) are shown. All data are presented as mean ± SD. ns, not significant, individual data points from independent biological replicates (n=6) are overlaid on the bars. ALB, albumin; ALP, alkaline phosphatase; ALT, alanine aminotransferase; AST, aspartate aminotransferase; CHE, cholinesterase; CK, creatine kinase; CRE, creatinine; GLB, globulin; HBDH, hydroxybutyrate dehydrogenase; LDH, lactate dehydrogenase; SD, standard deviation; UA, uric acid; UREA, urea.

Article Snippet: Fedratinib hydrochloride hydrate powder was purchased from TargetMol Chemicals Inc. (Boston, MA, USA).

Techniques: Derivative Assay, Control, Standard Deviation