Objective:To investigate the effect of prostaglandin E2 (PGE2) on expression and enzyme activity of matrix metalloproteinase 2(MMP2) by EP1 receptor in cholangiocarcinoma cells HuCCT1 cells. Methods:HuCCT1 cells were treated with PGE2, EP1 receptor agonist, EP1 receptor antagonist, protein kinase C(PKC) inhibitor and Ca2+ chelating agent, RT-PCR and gelatin zymography were employed to detect the mRNA level and enzyme activity of MMP2 in HuCCT1 cells. Results:The mRNA levels of MMP2 increased 89.14%(P < 0.01), 163.89%(P < 0.01); and the enzyme activity of MMP2 increased 69.11%(P < 0.05), 117.65(P < 0.01) compared with the control group after treated with 5 μmol/L PGE2, 5 μmol/L EP1 receptor agonist 17-PT-PGE2 respectively. The mRNA level and the enzyme activity of MMP2 in HuCCT1 cells treated with 10 μmol/L EP1 receptor antagonist sc-51322 decreased 47.74%(P < 0.05),84.58%(P < 0.01) compared with the cells treated with PGE2. After treated with 5 μmol/L PGE2 or 5μmol/L 17-PT-PGE2, the enzyme activity of MMP2 in EP1R-pcDNA3 transfected HEK293 cells increased 36.07%(P < 0.05), 61.59%(P < 0.05) compared with the control group. When treated with 5 μmol/L PKC inhibitor BIS-1, 10 μmol/L Ca2+ chelating agent BAPTA-AM, the mRNA levels of MMP2 decreased 44.17%(P < 0.05),34.42%(P < 0.05); and the enzyme activities of MMP2 decreased 70.95%(P < 0.05),71.82%(P < 0.05) compared with the cells treated with 17-PT-PGE2. Conclusion:PGE2 might up-regulate the mRNA level and the enzyme activity of MMP2 through EP1 receptor in cholangiocarcinoma HuCCT1 cells, which could be partly related to the Ca2+/PKC signaling pathway.