Objective To establish lentiviral expression vectors for Smurf1 silencing and assess the effects of Smurf1 silencing on cell migration. Methods HeLa and A549 cells were infected with lentiviral expression vectors for Smurf1 silencing respectively. After 7 days, the stable cell lines with Smurf1 silencing were obtained after puromycin-resistance screening, enrichment and expansion. The intracellular gene and protein levels of Smurf1 were detected by qPCR and western blot. Transwell assay was used to assess the effect of Smurf1 silencing on cell migration. Results The stable cell lines with Smurf1 silencing are constructed successfully. Silencing of Smurf1 down-regulated cell migration rate detected by Transwell assay. Conclusion Smurf1 promotes cell migration.