Figure 3.

F. nucleatum Induces Cancer Metastasis via the Autophagy Pathway. (A) Western blot analysis was performed in HCT116 cells cocultured with F. nucleatum (F01), the autophagy inhibitor CQ (20 µM) or PBS (control). (B-D) Transwell assays (B) were conducted with HCT116 cells cocultured with F. nucleatum (F01), CQ or PBS (control). The indicated migrated (C) and invaded (D) cells were quantified in five randomly selected fields, and the data are presented as the means ± SDs (**P < 0.01, and ***P < 0.001; unpaired Student's t-test). (E-F) The mRNA expression of E-cadherin (E) and Vimentin (F) was analyzed in HCT116 cells cocultured with F. nucleatum (F01), CQ or PBS (control) (*P < 0.05 and **P < 0.01; unpaired Student's t-test; the bars indicate the SD of three experiments) (G) Schematic of the experimental setup. (H-K) Representative colorectal tumors (red arrows) and H&E staining and FISH images of tumors in APC^Min/+ mice treated with F. nucleatum (F01), CQ or PBS control (I). Statistical analysis of mouse weights (H; two-way ANOVA combined with Bonferroni's post hoc test), tumor numbers (J; nonparametric Mann-Whitney test) and tumor sizes (K) in the different groups (n = 5-6 mice/group; *P < 0.05, **P < 0.01, and ***P < 0.001; the error bars indicate the SDs). (L) E-cadherin expression in tumors from APC^Min/+ mice treated with PBS (control), CQ, F. nucleatum (F01) or F. nucleatum(F01) + CQ was measured by immunohistochemical analysis. (M) Western blot analysis was performed with tissues from APC^Min/+ mice in the different groups.