Fig. 6: Aberrant bile acid metabolism promotes migration of invasive LUAD.

a Representative image of immunohistochemical staining of TGR5 and vimentin on 126 IAC patient tumor from S-I (n = 33), S-II (n = 63), and S-III (n = 30) subtypes. Scale bar, 100 μM. b Immunohistochemical (IHC) score of TGR5 and vimentin from patients in S-I (n = 33), S-II (n = 63), and S-III (n = 30) subtypes. A two-sided One-way ANOVA test was used. The centerline represents the median, and the box bounds represent the inter-quartile range. The whiskers span 1.5-fold the inter-quartile range. c Kaplan-Meier curves comparing the disease-free survival and overall survival in IAC patients with a high group (histoscore > 80 in TGR5 expression) versus low group (histoscore ≤ 80 in TGR5 expression). A two-sided log-rank test was used. d Kaplan-Meier curves comparing the disease-free survival or overall survival in IAC patients with a high group (histoscore > 20 in vimentin expression) versus low group (histoscore ≤ 20 in vimentin expression). A two-sided log-rank test was used. e The correlation plot of TGR5 with vimentin expression with significant Pearson’s correlation in TCGA LUAD dataset (n = 525). R, Pearson’s correlation coefficient. f, g Transwell migration assays were performed on H1299 cells treated with CA (100 μM), TCDCA (100 μM), and GCDCA (100 μM) or treated with CA (100 μM) and transfected with or without siTGR5. Representative images (left, scale bar, 100 μm) for three biological repeats and statistical analyses (right, n = 5) of the migrated cells are shown. siCtrl, siControl. Data represent the mean ± s.e.m. and One-way ANOVA followed by Tukey’s multiple comparison test was used. h Diagram showing the experimental design for in vivo metastasis assay (see Methods). i Lung metastasis assay of A549-luciferase cells treated with indicated bile acids (Vehicle, n = 7; CA, n = 8; GCDCA, n = 7; TCDCA, n = 5, example mice shown to left). j Quantification of the metastasis nodules on the pulmonary surface of each groups (Vehicle, n = 7; CA, n = 8; GCDCA, n = 7; TCDCA, n = 5). In i and j, data represent the mean ± s.e.m. and two-tailed Student’s t-test was used.