Objective To investigate the role of Krüppel-like factor 5 (KLF5) in the process of deoxycholic acid (DCA)-induced intestinal metaplasia of gastric mucosa and its potential mechanism. Methods The expressions of KLF5 and Wnt3a in GES-1 cells treated with different concentrations of DCA, normal gastric mucosa and intestinal metaplasia tissues were detected by Western blot and RT-PCR. GES-1 cells were divided into blank control group, DCA group, DCA+si-Cont group and DCA+si-KLF5 group. Then the cell proliferation ability and apoptosis were detected by MTT assay and flow cytometry, respectively. The expressions of TFF2, VIL1 and CDX2 were detected by Western blot and RT-PCR. Furthermore, the protein expression levels of Wnt3a, β-catenin and CDX2 in si-KLF5 transfected GES-1 cells were detected after treatment with LiCl (Wnt activator) by Western blot. Results The expression levels of KLF5 and Wnt3a were significantly increased in the intestinal metaplasia tissues compared with the normal gastric mucosa tissues, they were also increased gradually with the increase of DCA concentration in the GES-1 cells. In addition, the increased cell proliferation ability, the decreased cell apoptosis rate and the elevated expressions of TFF2, VIL1 and CDX2 were detected in the GES-1 cells after treatment with 500 μmol/L DCA. However, the cell proliferation was decreased, the cell apoptosis was increased and the expressions of TFF2, VIL1 and CDX2 were downregulated in the GES-1 cells after transfection with si-KLF5, which attenuated the effect of DCA. Furthermore, Wnt activator LiCl attenuated the effect of si-KLF5 on the expressions of Wnt3a, β-catenin and CDX2 in the DCA-induced cells. Conclusions KLF5 may promote the DCA-induced intestinal metaplasia of gastric mucosa by activating the Wnt pathway.