Journal: World Journal of Surgical Oncology

Article Title: FOXA2 promotes the occurrence and development of osteosarcoma cells by regulating ferroptosis through the Rheb/mTOR axis

doi: 10.1186/s12957-026-04308-4

Figure Lengend Snippet: a Nine common target genes were identified by overlapping the mTOR signaling pathway gene set with FOXA2 target genes predicted by CHIP-ATLAS, ENCODE, GTRD, and hTFtarget databases. b qRT-PCR analysis of EIF3A, EIF4A1, EIF4EBP1, mTOR, PI3KCA, Rheb, TSC1, RPS6KB1, and TSC2 in MNNG-HOS and 143B cells. c JASPAR database predicted binding sites between transcription factor FOXA2 and Rheb promoter. d ChIP assay was conducted to verify the FOXA2-binding site in the Rheb promoter region within 143B and MNNG-HOS cell lines. Controls comprised IgG and input fractions. e Luciferase reporter plasmids with either wild-type or mutant Rheb2 promoters were used for assessment of luciferase activities in 143B and MNNG-HOS. The mutation site is highlighted in red. f 143B and MNNG-HOS cells with FOXA2 knockdown, FOXA2 knockdown combined with Rheb overexpression, and negative controls were used to analyze Rheb, total and phosphorylated mTOR, xCT, and GPX4 at the protein level. Data are presented as mean ± SD. Significance levels were set as follows: ns p > 0.05, * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: The utilized antibodies included FOXA2 (1:500,Proteintech, Wuhan, China), GPX4 (1:1,000; Proteintech, Wuhan, China), xCT (1:1,000; Proteintech, Wuhan, China) (1:1000; Abclonal, Wuhan, China), mTOR (1:2,000; Proteintech, Wuhan, China), p-mTOR (1:2,000; Proteintech, Wuhan, China), Rheb (1:1,000; Abclonal, Wuhan, China), β-actin (1:10,000; Abclonal, Wuhan, China), HRP goat anti-mouse IgG (H + L) (1:5,000; Proteintech, Wuhan, China) and HRP-conjugated goat anti-rabbit IgG (H + L) (1:5,000; Proteintech, Wuhan, China).

Techniques: Quantitative RT-PCR, Binding Assay, Luciferase, Mutagenesis, Knockdown, Over Expression

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