Objective：To investigate the effect of suppressed expression of homeobox A6（HOXA6） on the proliferation，migration and invasion of HCT116 colorectal cancer cells. Methods：Recombinant plasmids were constructed and transfected via Lipofectamine 2000. Transfection and interference efficiency was detected by fluorescence microscope，RT-PCR and Western blot. Moreover，cell proliferation，migration and invasion were measured by CCK-8，colony formation and Transwell assays. The protein expression of E-cadherin，N-cadherin and Vimentin was identified by Western blot. Results：The transfection efficiency was nearly 30%. The expres－sion of HOXA6 mRNA（P=0.003） and protein（P=0.000） was significantly lower in all interference groups than in shRNA-NC group，and the most interference efficient plasmid was shRNA1-HOXA6（P=0.000）. CCK-8 showed that the A450 values in shRNA1-HOXA6 group were lower than in shRNA-NC group（P=0.005）. Colony formation revealed that the colonies in shRNA1-HOXA6 group （132±45） were considerably fewer than in shRNA-NC group（353±45）（P=0.004）. Besides，Transwell assay showed that the num－ber of migration and invasion cells was significantly fewer in shRNA1-HOXA6 group[（134±28），（60±4）] than in shRNA-NC group [（276±94），（168±50）] （P=0.025，P=0.008）. Moreover，the protein expression of E-cadherin was higher in shRNA1-HOXA6 group （0.490±0.025） than in shRNA-NC group（0.326±0.032）（P=0.003），but N-cadherin（0.112±0.016） and Vimentin（0.886±0.033） expression was lower in shRNA1-HOXA6 group than in shRNA-NC group[（0.147±0.013），（1.159±0.040）]（P=0.037，P=0.001）. Conclusion：Knockdown of HOXA6 expression can remarkably suppress the proliferation，migration and invasion of HCT116 colorectal cancer cells.