This study was to prepare the anionic liposomes which were to release anticancer drug (doxorubicin) at the hyperthermia temperature $({sim}42^{circ}C)$ and to stabilize in bovine serum solution at $37^{circ}C$. The vesicle size and zeta potential of liposomes in Tris-HCl buffered solution (pH 7.4) were measured by an electrophoretic light scattering spectrophotometer. To estimate the stability of liposomes, liposome size was measured in bovine serum solution at $37^{circ}C$ for 72 h. The release of doxorubicin from liposome was determined by measuring the fluorescence intensity using fluorescence spectrophotometry with temperature and time. The size of liposomes was from 120 to 160 nm and zeta potential was from $-33.3{pm}2.4$ to $-75.6{pm}6.9;mV$. Anionic liposome was stabilized in bovine serum solution at $37^{circ}C$ within 72 h. Additionally, the release transition temperature of doxorubicin from liposomes was increased by increasing mole % of anionic phospholipid.