Journal: Scientific Reports

Article Title: Cancer-Associated fibroblasts regulate the development of cholangiocarcinoma through IL-6/STAT3/AKR1C3 signaling axis

doi: 10.1038/s41598-026-37583-y

Figure Lengend Snippet: CAFs regulated the occurrence and development of CCA by activating the AKR1C3/STAT3 signaling axis. (A) The mRNA expression levels of AKR1C3 in QBC939 and TFK1 cells after co-incubation with CAFs were detected by Q-PCR. ** P < 0.01. (B) The protein expression levels of AKR1C3, P-STAT3, and T-STAT3 in QBC939 cells after co-incubation with CAFs at different times were measured by Western blot. (C) After CAF treatment with or without Ab-IL-6 and (or) AKR1C3 knockdown for 8 h, the expression of AKR1C3, P-STAT3, and T-STAT3 were detected by Western blot in QBC939 cells. (D) After treatment of CAFs with or without Ab-IL-6 and (or) AKR1C3 knockdown for 8 h, the expression of PCNA, P-GP, GLUT-1, and PFK-1 were detected by Western blot in QBC939 cells. The proliferation (E) and glycolysis levels (F) in QBC939 and TFK1 cells after treatment of CAFs with or without Ab-IL-6 and (or) AKR1C3 knockdown were assessed by CCK-8, glucose uptake, and lactate release, respectively. * P < 0.05, ** P < 0.01, *** P < 0.001. The cell colony (G) of QBC939 cells exposed to 40 µM 5-FU after treatment of CAFs with or without Ab-IL-6 and (or) AKR1C3 knockdown were detected by crystal violet staining. Ab-IL-6, 2ug/ml.

Article Snippet: Antibodies against P-STAT3 (9145 S), T-STAT3 (30835 S), PCNA (2586 S), P-GP (13879 S), GLUT-1 (73015 S) were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Expressing, Incubation, Western Blot, Knockdown, CCK-8 Assay, Staining

Journal: Scientific Reports

Article Title: Cancer-Associated fibroblasts regulate the development of cholangiocarcinoma through IL-6/STAT3/AKR1C3 signaling axis

doi: 10.1038/s41598-026-37583-y

Figure Lengend Snippet: CAFs regulated the occurrence and development of CCA via the IL-6/STAT3/AKR1C3 signaling axis in vivo. (A) shAKR1C3-QBC939 cells and control cells were subcutaneously injected in nude mice to establish xenograft tumors. The representative tumors and their volume are depicted graphically. (B) The growth and sensitivity to 5-FU were detected in the mixed xenografts of CAFs and shAKR1C3-QBC939 cells. (C) HE and IHC staining were performed to examine CAF-QBC939 and QBC939 xenografts. (D) The protein expressions of AKR1C3, P-STAT3, PCNA, P-GP, GLUT-1, and PFK-1 in the CAFs-shAKR1C3-QBC939 xenografts with or without the treatment of 5-FU were detected by Western blot. (E) Schematic summary illustrating how CAF-derived IL-6 activates the STAT3/AKR1C3 axis in cholangiocarcinoma cells to drive tumor proliferation, chemoresistance, glycolysis, and metastatic potential.

Article Snippet: Antibodies against P-STAT3 (9145 S), T-STAT3 (30835 S), PCNA (2586 S), P-GP (13879 S), GLUT-1 (73015 S) were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: In Vivo, Control, Injection, Immunohistochemistry, Western Blot, Derivative Assay