Objective:To investigate the expression level of PKM2 in cholangiocarcinoma (CCA)tissues,then study the effect of PKM2 down-regulation on migration,invasion and proliferation in cholangiocarcinoma cell lines. Methods:RNA and protein expressions of PKM2 in CCA tissues and paired adjacent tissues were detected by qRT-PCR and immunohistochemistry. PKM2 was down-regulated by a lentiviral vector expression system in cholangiocarcinoma cell lines HuCCT-1 and HCCC-9810. qRT-PCR and Western blot were performed to analyze the mRNA and protein expression of PKM2 in both cell lines. Cell migration, invasion and proliferation were assessed by wound-healing experiment, matrigel invasion and Cell Counting Kit-8(CCK-8). Results:The expression of PKM2 in CCA tissues had a higher level than that in paired adjacent tissues. The mRNA and protein expressions of PKM2 in the experimental group (PKM2-shRNA)were significantly lower than those in the two control groups,confirmed by qRT-PCR and Westen blot (P < 0.05). Compared to the empty vector group (PKM2-NC)and the normal control group (PKM2),the cell invasion,migration and proliferation were significantly decreased in the experimental group (PKM2-shRNA)(P < 0.05). Conclusion:Down-regulation of PKM2 by PKM2 shRNA can inhibit migration,invasion and proliferation of HuCCT-1 and HCCC-9810 cells.