The inhibiting effect of stachyose from Stachys floridana Schuttl.ex Benth on the proliferation of Caco-2 cell line and its potential mechanism were investigated.Caco-2 cells were incubated in serum-free media, the inhibiting effect of stachyose on Caco-2 cell proliferation were analyzed by MTT assay and the apoptosis of Caco-2 cells were determined by flow cytometry, then cell damage degree was studied by the detection of LDH release rate. The results showed that stachyose inhibited the proliferation of Caco-2 cell lines in a dose- and time- dependent manner. After pretreatment with different concentrations of stachyose (0.5, 1, 2, 4 mmol/L), cell apoptosis were obtained. The rise of LDH release rate indicated that stachyose inhibited the reduction ability of pyruvate to lactic acid, and limited the ability of gaining energy via glycolysis path way in Caco-2 cells, which suggests the inhibiting effect of stachyose on the proliferation of tumor cell may be relatedto hindering anaerobic glycolysis pathway and reducing the release of energy.