Fig. 6: FTD/TPI inhibits organoid viability and proliferation in colorectal cancer.

A Relative viability of colorectal cancer organoids derived from 12 patients was measured after treatment with FTD/TPI (0, 12.5, 25, 50 μM) for 7 and 14 days. B, C Accumulation levels of reactive oxygen species (ROS) and lipid peroxidation (MDA) were observed by fluorescence microscopy after treatment with FTD/TPI (0, 12.5, 25, 50 μM) for 7 days in colorectal cancer organoids. The first row represents green fluorescence images for ROS, red fluorescence represents MDA, the second row shows blue fluorescence images for DAPI, the third row presents merged fluorescence images, and the fourth row displays bright-field microscopy images of the organoids. Scale bar = 100 μm, magnification at 20X. D Microscopic images of colorectal cancer organoids from three patients were captured after treatment with 12.5 μM FTD/TPI and 1 μM Erastin for 7 days to observe and quantify organoid size. Scale bar = 200 μm, magnification at 4X and 20X. E Microscopic image of colorectal cancer organoid from one patient treated with 12.5 μM FTD/TPI and 20 μM Ferrostatin-1 for 7 days to observe and quantify organoid size. Scale bar = 200 μm. F Western blot analysis of p53 and SLC7A11 protein expression in colorectal cancer organoids from three patients after treatment with FTD/TPI (0, 12.5, 25, 50 μM) for 7 days. G Relative viability of colorectal cancer organoids was measured after treatment with FTD/TPI (0, 12.5, 25, 50 μM) in combination with 2 μM Erastin for 7 days in two patients. H Relative viability of colorectal cancer organoids was measured after treatment with FTD/TPI (0, 12.5, 25, 50 μM) in combination with 4 mM sulfasalazine for 7 days in nine patients. I, J Microscopic images of colorectal cancer organoids from patients were captured after treatment with 12.5 μM FTD/TPI and 4 mM sulfasalazine for 7 days to observe and quantify organoid size. Scale bar = 200 μm, magnification at 4X, 10X, 20X, and 40X. K Western blot analysis of SLC7A11 protein expression in colorectal cancer organoids from four patients after treatment with FTD/TPI (12.5 μM) in combination with sulfasalazine (4 mM) for 48 h. Data were presented as representative images or mean ± SD from at least three independent replicates. Statistical analysis was performed using a two-tailed paired t-test. *p < 0.05, **p ≤ 0.01, ***p < 0.001, ****p ≤ 0.0001, indicating statistical significance.