Figure 3.

PBC but not PSC mediates specific control of CCA outgrowth. (A) Experimental schedule in C57Bl/6 mice. 1 wk after the end of cholangitis induction (see Fig. 1, A and B), syngeneic cancer cells were engrafted through s.c. injection and tumor growth (TG) was followed up (B–D and H–J). Alternatively, mice received plasmids encoding the oncogenes NICD and constitutively active AKT by hydrodynamic injection in the tail vein, and their liver was collected 5 wk later (E–G). (B–D) Tumor growth of transplantable SB1-JP4 CCA tumors in control, PBC, and PSC mice. For control group, n = 7, and for PBC and PSC groups, n = 9. One representative experiment out of two is shown. (E and F) Photographic images of control (E) and PBC (F) mouse livers after NICD/AKT plasmid injection. Green arrows indicate some tumor nodules. Scale bars correspond to 1 cm. For control group, n = 9, and for PBC group, n = 10. (G) Rate of CCA incidence and number of tumor nodules per liver in control and PBC mice that received the oncogenes NICD and AKT. P values were calculated using a Fisher’s exact test or a Mann–Whitney U test for comparing the rate of CCA incidence and the number of tumor nodules, respectively. (H–J) Growth of transplantable HCC (H), NSCLC (I), and fibrosarcoma (J) tumors in control and PBC mice. (H) For control group, n = 5, and for PBC group, n = 7. (I) For each group, n = 5. (J) For each group, n = 4. Graphs show mean (±SEM; B and H–J) or individual (C and D) tumor growth curves. Individual curves of HCC, NSCLC, and fibrosarcoma growth are displayed in Fig. S1, D–F. Regarding the comparison of the tumor growth curves, tumor growth P values were calculated by means of a linear mixed-effects model in comparison to control group. Regarding the comparison of the rate of tumor-free (TF) mice, pairwise tumor-free P values were calculated with Fisher’s exact test in comparison to control group. *, P < 0.05; **, P < 0.01; ****, P < 0.0001. AKT, Protein kinase B; t.v., tail vein.