This article introduces the characteristics and establishment of MDCK-MDR1 cell line, and its applications as an in vitro model for the assessment of the membrane permeability properties of drugs. MDCK-MDR1 cell model is widely used in drug screenings, and the study of mechanisms of drug interaction and absorption or transport. The review summarizes the progress and applications of MDCK-MDR1 cell model. Because there is high polarized P-glycoprotein (P-gp) expression in MDCK-MDR1 cells, it could be used as a drug transport model to screen P-gp substrates or inhibitors, as well as models of intestinal mucosa, blood brain barrier and kidney. Although there are some disadvantages in MDCK-MDR1 cell model, it is an effective tool for implementing permeability of drug in an optimal fashion.