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纸质出版:2007
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陈姝,娄世锋,张颖,张萍.GM-CSF耦联米托蒽醌脂质体的制备及体外抑瘤作用的实验研究[J].陆军军医大学学报(原第三军医大学学报),2007,29(10):925-928.
CHEN Shu,LOU Shi-feng,ZHANG Ying,ZHANG Ping.Preparation of LEM-GM-CSF and its inhibition effect on HL60/ADM cells in vitro[J].J Amry Med Univ (J Third Mil Med Univ),2007,29(10):925-928.
目的 制备粒-巨噬细胞集落刺激因子 (granulocyte-macrophage colony stimulating factor,GM-CSF)耦联米托蒽醌脂质体(liposome-entrapped mitoxantrone,LEM),比较其与LEM和米托蒽醌(dihydroxyanthraquinone,DHAQ)对HL60/ADM细胞的体外杀伤作用。 方法 用逆向蒸发法制备LEM,高速离心法纯化LEM,比色法测定包封率,透射电镜下观察LEM的结构和粒径,以戊二醛交联法制备出LEM-GM-CSF,紫外分光光度法测定耦联率,流式细胞术测定LEM-GM-CSF中GM-CSF的活性保留率。通过MTT法检测所制备LEM-GM-CSF 、LEM 及DHAQ对HL60/ADM细胞的杀伤作用。 结果 LEM粒径分布均匀,为170~220 nm,包封率为80%,GM-CSF与LEM有较高的耦联率及活性保留率,分别为42.3%及74.6%;LEM-GM-CSF 、LEM 及DHAQ对HL60/ADM细胞具有杀伤作用,且LEM-GM-CSF的杀伤作用显著强于LEM和DHAQ。LEM-GM-CSF、LEM及DHAQ对HL60/ADM细胞作用24 h的IC
50
分别是8.73、12.42、27.31 μg/ml;作用48 h的IC
50
分别是:0.62、8.25、12.44 μg/ml。 结论 本研究制备的LEM-GM-CSF对 HL60/ ADM细胞具有明显的杀伤作用
有望成为良好的抗白血病药物新剂型。
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 lip
osome 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 IC
50
of HL-60/ADM was 8.73
12.42
27.31 μg/ml respectively and for 48 h the IC
50
were 0.62
8.25
12.44 μg/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.
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