Thymol has antioxidant, apoptotic, anti-cancer and, anti-inflammatory properties. In this study iron oxide nanoparticles were synthetized by chemical co-precipitation method, then coated with albumin and finally loaded with thymol. The properties and characteristics of these composites were investigated by x-ray diffraction (XRD), TEM, FTIR, FE-SEM, DLS, zeta potential techniques and, vibrating-sample magnetometry (VSM). Finally, the release and encapsulation behavior of these compounds was calculated. The anti-cancer effect of thymol loaded albumin-coated Fe3O4 nanoparticles were investigated by the MTT method. The rate of apoptosis was assessed by Flow Cytometry test, while BAX and BCL2 were assessed by ELISA test. Expression of the MAP1LC3A gene was also examined by the real-time PCR technique. The results of MTT, ELISA, Flow Cytometry, and, real-time PCR techniques showed that thymol loaded albumin-coated Fe3O4 nanoparticles at concentration of 150 & mu;g/mL for 72-hour treatment most significantly affected cytotoxicity, apoptosis and, autophagy. Hence, this compound probably exhibited anti-cancer effect on the MDA-MB-231 cell line by activating the autophagy and apoptosis pathways.