Tetrahydroxylated bile acids prevents malignant progression of Barret esophagus in vitro by inhibiting the interleukin-1β-nuclear factor kappa-B pathway

Figure 2 Effect of deoxycholic acid, tetrahydroxylated bile acids and their combination on adult tissue-resident stem cells isolated from Barrett esophagus clinical biopsies cytotoxicity. Representative green fluorescent protein-labeled (GFP) fluorescent images of Barrett esophagus clinical biopsies (BE-ASCs) in transwell inserts after 15 days in culture with various deoxycholic acid (DCA) and hydrophilic tetrahydroxylated bile acids (THBA) concentrations. 1 × 10⁶ BE-ASCs were plated in 12 Matrigel coated transwell inserts on the feeder layer of irradiated 3T3 mouse embryonic fibroblasts (1 × 10⁵ fibroblasts per transwell). A: BE-ASCs cultured with 100 μM DCA showed slight cytotoxicity and significant decreasing of viability (22-fold) after treatment with 150 μM DCA or higher; B: BE-ASCs treated with 3000 μM THBA or higher demonstrated significant decreases in viability; C: BE-ASCs co-treated with 2000 μM THBA/150 μM DCA showed 17-fold higher viability in comparison to cells treated with DCA alone; D-F: Viability levels of BE-ASCs treated with DCA (D), THBA (E), their combination (F), were correlated with GFP fluorescence intensity measured using ImageJ software. Averages from triplicates are presented as mean ± SD (n = 3). ^cP < 0.001. ^dP < 0.0001. DCA: Deoxycholic acid; THBA: Tetrahydroxylated bile acids.