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Radiofrequency (RF) hyperthermia has been widely used for tumor ablation since magnetic-fluid-hyperthermia (MFH) can be utilized for increasing temperature in tumor-region as a complementary-method for hyperthermia. In this study, the effectiveness of using the magnetite-nanoparticles (Fe₃O₄) in RF hyperthermia for breast cancer (BC) treatment by determining 3D-temperature-distribution using bioheat-transfer-mapping was evaluated. A breast-phantom with a tumor region was placed in an RF-device with 13.56 MHz frequency in different states (with and without-nanomagnetite). Parallelly, the calculations of the RF-wave and bioheat-equation were accomplished by numerical-simulation and finite-element-method (FEM) in COMSOL-software. The temperature differences were experimentally measured at different points of the phantom with a precision of 0.1 °C, with temperature of 3.6 °C and 6.1 °C in without and with nanomagnetic conditions in tumor area, respectively, and also for normal area with temperature of 1.8 °C and 1.9 °C in non-presence and presence states of 0.05 gr magnetite for both conditions, respectively. Moreover, the difference between the simulation and the experimental results was 0.54-1.1 %. The conformity between temperature measurement in experimental and simulation studies in tumor and normal areas showed the effectiveness of the application of MNPs for RF hyperthermia in tissue equivalent breast phantom. Finally, the positive effect of 0.05 gr of MNPs on BC treatment was confirmed.