SOX2 overexpression correlates with aggressive behavior and poor overall survival in cholangiocarcinoma (CCA). However, the cellular functions and precise role of SOX2 in CCA have not been elucidated. Here, we inserted SOX2 coding sequence to establish a CAA cell line which stably overexpressed SOX2. In vitro experiments showed that overexpression of SOX2 in cells was associated with increased cell proliferation, suppressed cell apoptosis, as well as enhanced cell migration and invasion. Our findings may lead to a better understanding of the biological effect of SOX2 and provide mechanistic insights for developing potential therapeutic strategies for CCA treatment.