Objective:To establish a MDA-MB-231 human breast cancer cell line stably express GFP-C-Daam1, and to investigate the influence of Daam1 on the mobility of breast cancer cells. Methods:The lentivrial vectors containing GFP or GFP-C-Daam1 were constructed and infected human breast cancer cells MDA-MB-231. MDA-MB-231 cells were screened by puromycin and stable expression of GFP or GFP-C-Daam1 was confirmed by Western blotting. Boyden chamber assay was used to test the intrinsic motility of the cells. Results:The cell lines stably expressing GFP or GFP-C-Daam1 were established successfully by the lentivrial infection. The GFP-C-Daam1 expressing cells showed greater intrinsic motility, suggesting the products of the interesting gene were active and functional. Conclusion:MDA-MB-231 breast cancer cell lines stably expressing GFP or GFP-C-Daam1 were successfully established using lentiviral methods. These cell lines can be used as model cells for further study of active Daam1 targeting.