Abstract: Gambogic acid(GA)-loaded reconstituted high-density lipoprotein nanoparticles(GA-rHDL-NPs)were prepared by sodium cholate method. The physicochemical properties such as morphology, particle size, Zeta potential, entrapment efficiency and drug-loading capability were evaluated. Dialysis method was employed to investigate in vitro release of GA-rHDL-NPs. In addition, the cellular uptake experiments were performed to evaluate the tumor cellular targeting ability of GA-rHDL-NPs, and safety evaluation following the intravenous injection of GA-rHDL-NPs was carried out using hemolysis testing and irritation testing upon rabbit ear vein. It was found that the well-formulated GA-rHDL-NPs displayed regular spherical shape with particle size of(113. 53±2. 50)nm, Zeta potential of -(29. 48±0. 05)mV, entrapment efficiency of(95. 30±0. 37)% and drug-loading of(8. 51±0. 95)%. The GA-rHDL-NPs solution was found to be stable after one-month storage at 4 °C. In vitro 24-h and 72-h accumulative releases of GA from GA-rHDL-NPs were approximately 24. 3% and 73. 6%, respectively. It revealed that there was much more uptake of GA-rHDL-NPs by HepG2 cells than by L02 cells. GA-rHDL-NPs also exhibited less local venous irritation and hemolysis than did the unencapsulated drug group. Therefore, the GA-rHDL-NPs possessed accepted entrapment efficiency, stability, sustained-release, dispersion and safety properties, and tumor cell uptake, which might be suitable for iv injection and cancer therapy.