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

Journal: International Journal of Biological Sciences

Article Title: GLUT3 drives paclitaxel resistance in peritoneal metastatic gastric cancer by promoting H3K18 lactylation-mediated MAPKAP1 transcription to suppress ferroptosis

doi: 10.7150/ijbs.130059

Figure Lengend Snippet: GLUT3 inhibition sensitizes GC cells to paclitaxel in the context of peritoneal metastasis by promoting ferroptosis. A) GC cells were treated with saline, paclitaxel (AGS/HGC27: 0.1 μM; MKN45: 10 μM), a GLUT inhibitor-1 (1 μM), a combination of GLUT inhibitor-1 and paclitaxel and a combination of GLUT inhibitor-1, paclitaxel and Fer-1 (2 μM) for 24 h. Subsequently, cell viability was measured with a CCK8 assay. B) Representative in vivo bioluminescence images of nude mice before and after treatment. Mice were intraperitoneally injected with MKN45-GLUT3 cells (1×10⁷ cells in 200 μL of PBS). The pretreatment images were captured at 21 days post-injection. Posttreatment images were taken on day 35 (24 h after the last treatment) with the indicated agents: saline, paclitaxel, GLUT inhibitor-1, and a combination of GLUT inhibitor-1 and paclitaxel. C) Gross morphology of peritoneal metastatic nodules at the experimental endpoint (day 35): representative specimens from the saline, paclitaxel (15 mg/kg), GLUT inhibitor-1 (10 mg/kg), and combination therapy groups. The red arrows indicate gastric cancer peritoneal metastases. D) Weight quantification of peritoneal metastatic tumors. E) The levels of MDA in tumor tissues from the indicated groups were determined. F) Representative H&E staining and IHC staining images of GPX4, SLC7A11, and MAPKAP1 in tumor tissues from the indicated groups are shown. Scale bar: 100 μm and 50 μm. G) The ratio of GSH/GSSG in tumor tissues from the indicated groups was determined. H) Quantification of the protein expression of GPX4, SLC7A11, and MAPKAP1 in tumor tissues from the indicated groups (n=3). All the data are presented as the mean ± SD. *P<0.05 , ***P<0.001 , ****P<0.0001 , ns: not significant. Immunohistochemical staining was quantified using the H-score, calculated as staining intensity (0-3) × percentage of positive cells (1-4). The staining intensity was graded as follows: 0 (negative), 1 (weak), 2 (intermediate), or 3 (strong). The proportions of positively stained cells were categorized as follows: 1 (0-25%), 2 (26-50%), 3 (51-75%), or 4 (76-100%). Abbreviations: H&E: Hematoxylin and Eosin staining; IHC: Immunohistochemical; CCK8: Cell Counting Kit-8; Fer-1: Ferrostatin-1; MDA: Malondialdehyde; GSH: Glutathione (reduced); GSSG: Glutathione Disulfide (oxidized).

Article Snippet: To elucidate the role of GLUT3 in peritoneal metastasis and validate whether the combination of GLUT inhibitor-1 (HY-139605; MedChemExpress) and paclitaxel increases antitumor activity by promoting ferroptosis, two independent animal experiments were conducted.

Techniques: Inhibition, Saline, CCK-8 Assay, In Vivo, Injection, Staining, Immunohistochemistry, Expressing, Immunohistochemical staining, Cell Counting