Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Pharmacological induction of acetyl-CoA carboxylase 1 autophagic degradation attenuates lipid accumulation and cholangiocarcinoma progression

doi: 10.1186/s13046-025-03564-8

Figure Lengend Snippet: WA significantly attenuates lipid accumulation in CCA. A UMAP visualization of integrated scRNA-seq data (20 tumors, 12 adjacent tissues) showing seven major cell lineages, and malignant cholangiocyte identification by CopyKat analysis (dark blue: malignant, yellow: non-malignant). B Metabolic pathway enrichment in malignant versus non-malignant cholangiocytes. C Single-cell flux estimation analysis (scFEA) was performed to infer metabolic fluxes at single-cell resolution, revealing differential metabolite profiles and metabolic activities between malignant and non-malignant cholangiocytes. D CK19 IHC staining and oil red O staining to detect lipid accumulation in human CCA tissues, scale bar = 20 μm. E CK19 IHC staining and oil red O staining to detect lipid accumulation in the KRAS/P19 CCA model, scale bar = 20 μm. F CK19 IHC staining and oil red O staining to detect lipid accumulation in the YAP/AKT CCA model, scale bar = 20 μm. G The schematic for screening of anti-tumor inhibitors targeting lipid accumulation in a natural compound library containing 994 compounds. H The inhibitory rate of each inhibitor on HuCCT1 and RBE was detected by CCK8 assay. Red coloration indicated inhibitors that have achieved more than 50% inhibition. I BODIPY493/503 fluorescence staining and flow cytometry experiments were performed to detect lipid droplet content in HuCCT1 and RBE after treatment with each of the 24 compounds. J Molecular structure of WA. K This study employed a combination of BODIPY 493/503 fluorescence staining and Oil Red O staining to detect lipid droplet content in RBE and HuCCT1 cells 24 h after WA treatment. scale bar = 100 μm. L - M Lipids in WA treatment CCA cells were measured by LC-MS. The heatmap highlighted relative changes for each type of lipid metabolite with their individual average ion counts normalized to NTC. Data are representative of three independent experiments. scale bar = 100 μm. The data are represented as the mean ± SD of three independent experiments. * P < 0.05; ** P < 0.01; *** P < 0.001; ns, no significance

Article Snippet: For BODIPY 493/503 staining: 100 μL of 10 μM BODIPY 493/503 working solution (MCE, NJ, USA) was added to each well and incubated for 30 min.

Techniques: Immunohistochemistry, Staining, Drug discovery, CCK-8 Assay, Inhibition, Fluorescence, Flow Cytometry, Liquid Chromatography with Mass Spectroscopy