Preparation of LEM-GM-CSF and its inhibition effect on HL60/ADM cells in vitro / 第三军医大学学报

ABSTRACT

Objective To prepare liposome-entrapped mitoxantrone (LEM)-GM-CSF and observe the cytotoxicity of HL-60/ADM cells treated with LEM-GM-CSF, LEM and dihydroxyanthraquinone (DHAQ) in vitro. Methods LEM was prepared by reverse phase evaporation (REV). High speed centrifugation was applied to separate LEM and dissociate DHAQ. Colorimetry was employed to determine encapsulation efficiency. The liposome structure and particle size were determined by transmission electron microscopy. GM-CSF was coupled to LEM by glutaraldehyde method. UV-spectrophotometric analysis was applied to measure the coupled efficiency. Flow cytometry was applied to determine the immunoconjugate retained efficiency. The cytotoxicity of HL60/ADM cells and interdiction efficiency of GM-CSF were investigated by MTT test. Results The encapsulation efficiency of LEM was 80%. Most liposomes were monolayer, and the particle size was 170-220 nm. Its coupled efficiency with GM-CSF was 42.3%, and the immunoconjugate retained efficiency was 74.6%. All LEM-GM-CSF, LEM and DHAQ had cytotoxicity on HL60/ADM, their cytotoxic power in decrement sequence LEM-GM-CSF, LEM, DHAQ. After treated with LEM-GM-CSF, LEM and DHAQ for 24 h, the IC50 of HL-60/ADM was 8.73, 12.42, 27.31 ?g/ml respectively and for 48 h the IC50 were 0.62, 8.25, 12.44 ?g/ml. The inhibition rate increased in a dose-dependent manner. Conclusion The encapsulation efficiency, the coupled efficiency and the immunoconjugate retained efficiency of LEM-GM-CSF prepared by our method were satisfying. LEM-GM-CSF representing anti-leukaemia efficiency in vitro had cytotoxicity on HL60/ADM cells.