To evaluate the expression of ANAX1 in the different stages of CCA tissues in O. viverrini + NDMA-induced CCA in the hamster model, immunohistochemistry was used to evaluate the inflamed tissues at 21 d, when the tumors began at 3 mo, and tumor progression at 6 mo post-treatment. The percentage of CCA 60% (3/5) at 3 mo, and 100% (5/5) at 6 mo post-treatment were described previously[23]. Here, it was shown that the expression of ANXA1 was observed mainly in the cytoplasm of the epithelial bile ducts, some inflammatory cells (large cell, macrophage-like cell), and tumor cells which increased with time as bile duct proliferation and CCA development progressed (Figure (Figure4).4). The level of its expression at 21 d was +, 3 mo was ++, and 6 mo was +++ for all five animals per group. In addition, low or a little expression was found in the normal livers with no changes in each time-sacrificed group.