Objective To investigate the effect of rhodioside on the proliferation, invasion and migration of human osteosarcoma 143B cells and its possible mechanism. Methods Osteosarcoma 143B cells were cultured in dulbeeeo modified eagle medium (DMEM) medium (including 10% serum and 1% double resistance). Osteosarcoma 143B cells were divided into control group, 30 and 60 μmol/L rhodioside treatment group. Cell proliferation was tested by methyl thiazol tetrazolium (MTT) assay at 24, 36, 72 h, Transwell method was used to detect three groups of cell invasion; migration ability were detected by cell scratch assay. After 24 h treatment, calculation of cell migration rate. After 24 h treatment, reverse transcriptase-polymerase chain reaction (RT-PCR) was used to detect the relative expression level of matrix metalloproteinase (MMP)-2 and MMP-9 mRNA. The protein expression level of MMP-2 and MMP-9 in each group was detected by Western blotting. The autophagy after 24 h was detected by transmission electron microscopy. Results After the treatment of 24 h, the absorbance value of 60 mol/L group (0.038±0.002) and 30 mol/L group (0.054±0.002) 143B cells was lower than that of the control group. Compared with the control group, rhodioside with concentration of 30 and 60 mol/L at 48 h and 72 h inhibited the proliferation of 143B cells, respectively, with a dose-dependent and time dependent manner. The number of transmembrane of 143B cells in 60 μmol/L (48.7±8.2) and 30 μmol/L (76.8±12.8) rhodioside group was less than that of the control group (94.5±11.9), t1=8.056, P=0.000; t=2.472, P=0.038, respectively. The migration rates of 143B cells in 60 μmol/L [(52.9±6.8)%] and 30 μmol/L [(82.6±3.7)%] rhodioside group were lower than those of the control group [(98.7±0.3)%, t=12.576, P=0.000, t=9.794, P=0.000], respectively. The relative expression of protein and mRNA of MMP-2 and MMP-9 in 60 mol/L and 30 mol/L rhodioside treatment group was lower than that of the control group. The number of autophage in the 60 mol/L rhodioside group was (2.6±0.5) more than that of the control group (1.3±0.4, t=3.517, P=0.025). Conclusion Rhodioside inhibits the proliferation, migration and invasion of osteosarcoma cells, and its mechanism may be related to the induction of autophagy and the inhibition of the expression of MMP family protein. Key words: Osteosarcoma; Rhodioside; Migration; Invasion; Autophagy