Abstract: Objective To study the effects of honokiol on proliferation, migration and apoptosis of human tongue carcinoma CAL-27 cells. Methods Routinely cultured CAL-27 cells were treated with 20, 40, or 60 μmol/L honokiol and the changes in cell proliferation were assessed with MTT assay. The scratch wound healing assay was used to assess the migration ability of the treated cells, and the cell apoptosis was detected with Hoechst33342 fluorescence staining and annexin V-FITC/PI method. The protein expression levels of p-Pi3k, p-Fak, Fak, MMP-2, MMP-9, p-Akt, Akt, Bax, Bcl-2 and cleaved-caspase-3 in the treated cells were detected using Western blotting. Results Treatment with honokiol at 20, 40, and 60 μmol/L for 24 h significantly lowered the proliferation and migration ability of CAL-27 cells. The number of apoptotic cells increased with the increase of honokiol concentration, which resulted in a cell apoptosis rate of (15.24±2.06)% at 20 μmol/L, (35.03±2.42)% at 40 μmol/L, and (48.13±4.61)% at 60 μmol/L, as compared with (6.53±1.80)% in the control group. The expressions of p-Pi3k, p-Fak, MMP-2, MMP-9, p-Akt and BCL-2 decreased and those of Bax and cleaved-caspase-3 increased significantly in the cells after the treatment (P<0.01). Conclusion Honokiol can inhibit the proliferation and migration and induce apoptosis of CAL-27 cells in vitro possibly by regulating the expressions of p-Pi3k, p-Fak, MMP-2, MMP-9, p-Akt, Bax, Bcl-2 and cleaved-caspase-3.