Journal: Acta Pharmaceutica Sinica. B

Article Title: Targeting cancer-associated fibroblast-activated HGF/c-MET pathway inhibits extrahepatic cholangiocarcinoma progression and restores gemcitabine therapeutic sensitivity

doi: 10.1016/j.apsb.2026.02.023

Figure Lengend Snippet: Effect of CAFs on eCCA progression. (A) Immunofluorescence staining of CAFs/NFs showed positive expression of α -SMA, Vimentin and FAP. Scale bar: 200 μm. (B) Effect of CAF-CM/NF-CM on proliferation of TFK-1 and CBC3T-1 cells. Negative control (NC) ( n = 3). (C, D) Effect of CAF-CM/NF-CM on migration and invasion of TFK-1 and CBC3T-1 cells ( n = 3). Scale bar: 200 μm. (E, F) Representative fluorescent images of PDOs monocultures or co-cultures with CAFs/NFs and quantitative analysis of total organoid area ( n = 3). PDOs (green), CAFs/NFs (red). Scale bar: 200 μm. (G) Representative tumor images of cell-derived xenografts: cancer cells, cancer cells + NFs and cancer cells + CAFs. (H, I) Mean tumor volume and tumor weight ( n = 8). (J) Representative images of H&E and IHC staining of α -SMA, CK7 and Ki67 ( n = 8). Data are shown as mean ± SD. ns, no significance; ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001; ∗∗∗∗ P < 0.0001.

Article Snippet: The human eCCA TFK-1 cell line was procured from Creative Bioarray (NY, USA).

Techniques: Immunofluorescence, Staining, Expressing, Negative Control, Migration, Derivative Assay, Immunohistochemistry

Journal: Acta Pharmaceutica Sinica. B

Article Title: Targeting cancer-associated fibroblast-activated HGF/c-MET pathway inhibits extrahepatic cholangiocarcinoma progression and restores gemcitabine therapeutic sensitivity

doi: 10.1016/j.apsb.2026.02.023

Figure Lengend Snippet: Inhibition of HGF/c-MET signaling pathway suppresses CAFs-induced eCCA progression. (A) Representative IHC images of c-MET expression in eCCA ( n = 27) and para-tumor tissues ( n = 9). (B) IHC staining intensity of c-MET in eCCA tissues ( n = 27) and para-tumor tissues ( n = 9) of TMAs. (C) Representative images of multiplex immunofluorescence staining ( α -SMA, green; p-c-MET, yellow; CK19, red) in eCCA and para-tumor tissue. (D) Percentage of p-c-MET + CK19 + cells to CK19 + cells in eCCA tissues and para-tumor tissues. (E) Mean fluorescence intensity of α -SMA + in eCCA and para-tumor tissues. (F) Western blot analysis of p-c-MET, total c-MET, p-PI3K, total PI3K, p -AKT, and total AKT in TFK-1 and CBC3T-1 cells treated with recombinant HGF at different time points (15, 30, 60, 120 min). (G, H) Western blotting of the c-MET/PI3K/AKT signaling pathway in TFK-1 and CBC3T-1 cells under the following conditions: normal control (NC), CAF-CM, CAF-CM supplemented with an HGF-neutralizing antibody (CAF-CM + HGF Ab), or recombinant HGF ( n = 3). (I, J) Western blot analysis of c-MET/PI3K/AKT pathway activity in eCCA cells treated with CAF-CM, with or without the c-MET inhibitors JNJ-38877605 or crizotinib ( n = 3). Data are shown as mean ± SD. ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001; ∗∗∗∗ P < 0.0001.

Article Snippet: The human eCCA TFK-1 cell line was procured from Creative Bioarray (NY, USA).

Techniques: Inhibition, Expressing, Immunohistochemistry, Multiplex Assay, Immunofluorescence, Staining, Fluorescence, Western Blot, Recombinant, Control, Activity Assay

Journal: Acta Pharmaceutica Sinica. B

Article Title: Targeting cancer-associated fibroblast-activated HGF/c-MET pathway inhibits extrahepatic cholangiocarcinoma progression and restores gemcitabine therapeutic sensitivity

doi: 10.1016/j.apsb.2026.02.023

Figure Lengend Snippet: Targeting the HGF/c-MET signaling pathway enhances gemcitabine treatment sensitivity in vitro . (A) Cell viability of TFK-1 and CBC3T-1 cells treated with gemcitabine or gemcitabine combined with HGF ( n = 3). (B) Representative images of the sensitivity of gemcitabine to PDOs monoculture systems or direct co-culture systems with CAFs ( n = 3). PDOs (green), CAFs (red). The percentage of total area of the PDOs was quantified and the viability was determined. Scale bar: 500 μm. (C) Schematic diagram for the construction of TFK-1 gemcitabine-resistant cells (TFK-1R). (D) Western blot analysis of c-MET protein levels in parental TFK-1 cells and gemcitabine-resistant TFK-1 cells (TFK-1R). (E) Cell viability of 100 μmol/L gemcitabine-treated TFK-1 and TFK-1R cells ( n = 3). (F) Cell viability of TFK-1R cells treated with 100 μmol/L gemcitabine or in combination with c-MET inhibitor under conditions of added HGF ( n = 3). (G) Dose–response curves of gemcitabine alone or in combination with c-MET inhibitors for treatment of TFK-1 and CBC3T-1 cells under conditions of added HGF ( n = 3). (H) Western blot analysis of c-MET protein levels in PDOs established from 10 cases of eCCA. (I, J) Representative images and dose–response curves of gemcitabine alone or in combination with c-MET inhibitor in a direct co-culture system of PDOs and CAFs. Patient 8 (P8) ( n = 3). Scale bar: 500 μm. Data are shown as mean ± SD. ns, no significance; ∗∗ P < 0.01; ∗∗∗ P < 0.001.

Article Snippet: The human eCCA TFK-1 cell line was procured from Creative Bioarray (NY, USA).

Techniques: In Vitro, Co-Culture Assay, Western Blot