Journal: Cancers

Article Title: OTOF Promotes Clear Cell Renal Cell Carcinoma Progression and Angiogenesis Through AKT-Dependent HIF/VEGFA Signaling

doi: 10.3390/cancers18152498

Figure Lengend Snippet: OTOF knockdown attenuates angiogenesis-related phenotypes and AKT/HIF/VEGFA signaling in ccRCC. ( A , B ) Schematic illustration and Western blot analysis showing VEGFA expression in conditioned medium derived from Vector and shOTOF 786-O cells. ( C ) ELISA quantification of VEGFA secretion levels in conditioned media from Vector and shOTOF ccRCC cells. ( D ) HUVEC viability assay showing that conditioned media from OTOF-knockdown 786-O and Caki-1 cells significantly suppressed endothelial cell viability. ( E ) Western blot analysis of AKT, p-AKT, HIF (Caki-1: HIF-1α; 786-O: HIF-2α) and VEGFA expression in 786-O and Caki-1 cells treated with Vector, Vector plus the AKT inhibitor MK-2206, shOTOF, or shOTOF plus the AKT activator SC79. ( F ) qPCR analysis of relative mRNA expression of HIF-1α in 786-O and Caki-1 cells, respectively. ( G ) qPCR analysis of relative mRNA expression of HIF-2α in 786-O and Caki-1 cells, respectively. ( H ) Western blot analysis of angiogenesis-related proteins in 786-O xenograft tumors. OTOF knockdown decreased the expression of CD31 and VEGFA, whereas SC79 treatment partially rescued their expression. ( I ) Proposed mechanism by which OTOF promotes angiogenesis in ccRCC through activation of the AKT/HIF/VEGFA signaling axis. ns: not significant ( p > 0.05), * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

Article Snippet: For AKT inhibition, cells were treated with MK-2206 (HY-10358, MCE, USA) at 1 μM for 3 h to assess AKT phosphorylation and at 1 μM for 124 h to examine downstream HIF/VEGFA signaling.

Techniques: Knockdown, Western Blot, Expressing, Derivative Assay, Plasmid Preparation, Enzyme-linked Immunosorbent Assay, Viability Assay, Activation Assay

Journal: Frontiers in Oncology

Article Title: Hypoxia induced DNMT3B and SHP2 signaling promoted HCC via suppressing P53 and MYH11 protein expression

doi: 10.3389/fonc.2026.1794481

Figure Lengend Snippet: Regulation of hepatocellular carcinoma progression by CBFβ-MYH11 under hypoxic conditions through AKT and SHP2 pathways. (A) Western blot experiments were performed to detect p-AKT, p-SHP2, p-DNMT3B, P53 and MYH11 protein banding plots and relative protein expression statistics in Normoxic group, Hypoxic group, Normoxic+MK-2206 group, Hypoxic+MK-2206 group, Normoxic+Batoprotafib group and Hypoxic+Batoprotafib group; (B) Methylation-qPCR assay was performed to detect the methylation levels of P53 and MYH11 in Normoxic group, Hypoxic group, Normoxic+MK-2206 group, Hypoxic+MK-2206 group, Normoxic+Batoprotafib group and Hypoxic+Batoprotafib group; (C) RT-qPCR experiments were performed to detect the relative mRNA expression of P53β and Δ133P53 in Normoxic group, Hypoxic group, Normoxic+MK-2206 group, Hypoxic+MK-2206 group, Normoxic+Batoprotafib group and Hypoxic+Batoprotafib group Statistical plots of the amount of P53β and Δ133P53. Data were expressed as mean ± standard deviation. N = 3; *P<0.05; **P<0.01; ns P>0.05: There was no significant difference.

Article Snippet: Akt inhibitor MK-2206 (5 nM), SHP2 inhibitor Batoprotafib (0.005 μM), P53 inhibitor Pifithrin-α (10 μM), and P53 activator Tenovin-1 (10 μM) were purchased from MCE.

Techniques: Western Blot, Expressing, Methylation, Quantitative RT-PCR, Standard Deviation

Journal: Frontiers in Oncology

Article Title: Hypoxia induced DNMT3B and SHP2 signaling promoted HCC via suppressing P53 and MYH11 protein expression

doi: 10.3389/fonc.2026.1794481

Figure Lengend Snippet: Migration, invasion, and angiogenesis of hepatocellular carcinoma under hypoxia induced by CBFβ-MYH11 through AKT and SHP2 pathways. (A) Western blot experiments were performed to detect the strip maps of MMP2 and HIF1α in the Normoxic group, Hypoxic group, Normoxic+MK-2206 group, Hypoxic+MK-2206 group, Normoxic+Batoprotafib group, and Hypoxic+Batoprotafib group and the relative protein expression statistics; GAPDH as control protein; N = 3; (B) Graphs of experimental results of Transwell assay to detect the cell migration ability of Normoxic group, Hypoxic group, Normoxic+MK-2206 group, Hypoxic+MK-2206 group, Normoxic+Batoprotafib group, and Hypoxic+Batoprotafib group as well as statistical graphs of the number of migrated cells; N = 3; (C) Plots of experimental results of cell scratch assay to detect cell migration ability of Normoxic group, Hypoxic group, Normoxic+MK-2206 group, Hypoxic+MK-2206 group, Normoxic+Batoprotafib group, and Hypoxic+Batoprotafib group as well as statistical plots of cell spacing; N = 3; (D) Graphs of the experimental results of Angiogenesis experiment to detect the angiogenic ability of Normoxic group, Hypoxic group, Normoxic+MK-2206 group, Hypoxic+MK-2206 group, Normoxic+Batoprotafib group, and Hypoxic+Batoprotafib group, as well as statistical graphs of the number of blood vessels; N = 3; (E) The results of HE staining experiments were plotted as well as the statistics of the number of blood vessels. N = 8; Data were expressed as mean ± standard deviation. **P<0.01; ns P>0.05: There was no significant difference.

Article Snippet: Akt inhibitor MK-2206 (5 nM), SHP2 inhibitor Batoprotafib (0.005 μM), P53 inhibitor Pifithrin-α (10 μM), and P53 activator Tenovin-1 (10 μM) were purchased from MCE.

Techniques: Migration, Western Blot, Stripping Membranes, Expressing, Control, Transwell Assay, Wound Healing Assay, Staining, Standard Deviation