The present study aimed to investigate the effect and mechanism of γ-aminobutyric acid (GABA) on the bio-nomics of cholangiocarcinoma QBC939 cells. A transwell cell culture chamber assay in vitro was used to detect the ability of invasion and metastasis of cholangiocarcinoma QBC939 cells. The expression of matrix metalloproteinase (MMP) mRNA and the enzymatic activity in QBC939 cells were detected by reverse transcription-polymerase chain reaction and gelatin zymography, respectively. Different concentrations of GABA inhibited the ability of the tumor cells in trans-Matrigel migration (100 μmol/l GABA, 60±9.54 vs. 42.67±4.04, P<0.05), and inhibited the activity and expression of MMP-2 and MMP-9. GABA inhibited the invasion and metastasis of cholangiocarcinoma QBC939 cells, and the mechanism may involve the down-regulation of the activity of MMPs.