The paper aims to compare zeta potentials, magnetic properties, electron spin resonance, photoluminescence (PL) spectra and antitumor effect of magneto-mechano-chemically synthesized magneto-sensitive nanocomplexes loaded with the anticancer drug doxorubicin (DOXO) during nanotherapy of Walker-256 carcinosarcoma carried out by a magnetic resonance system. Diamagnetic DOXO acquired the properties of a paramagnetic substance after synthesis. MNC comprising superparamagnetic nanoparticles (NP) and DOXO had different g-factors, zeta potentials, a lower saturation magnetic moment, area of the hysteresis loop, and a higher coercivity compared to similar MNC with ferromagnetic NP. The main PL peak of MNC spectrum was defined by DOXO at 598 nm. MNC composed of superparamagnetic NP and DOXO showed a lower standard deviation of the normal PL spectral distribution than MNC based on ferromagnetic NP in relation to conventional DOXO. MNC containing superparamagnetic NP responded to resonance conditions leading to a more pronounced antitumor effect compared to MNC with ferromagnetic NP in the course of magnetic nanotherapy for Walker-256 carcinosarcoma bearing animals (temperature inside the tumor did not exceed 40 °C). Therefore, these findings are associated with differences in chemotherapeutic effect between MNC due to a different surface charge and conformational changes in DOXO molecules during its magnetoelectric interaction with single- and multidomain NP.