应用平阳霉素与单克隆抗体Fab′片段偶联物抑制肝癌生长与肿瘤肝转移  

Inhibition of the growth of hepatoma and hepatic metastasis by pingyangmycin conjugated with Fab′fragment of monoclonal antibody

作　　者：刘小云[1] 尚伯扬[1] 刘秀均[1] 甄永苏[1] 

机构地区：[1]中国医学科学院中国协和医科大学医药生物技术研究所,北京100050

出　　处：《中华医学杂志》2001年第4期201-204,共4页National Medical Journal of China

基　　金：国家"九五"科技攻关基金!资助项目 (96 C0 2 0 1 15 )

摘　　要：目的　研制一种具有抑制肝癌生长和肿瘤肝转移作用的单克隆抗体Fab′片段与平阳霉素 (PYM)偶联物。方法　以胃蛋白酶酶解结合二巯基苏糖醇 (DTT)还原法制备单抗Fab′片段 ;以葡聚糖T 40 (dextranT 40 )为中介体制备Fab′片段和PYM偶联物Fab′ PYM ;用间接ELISA法、2 ,3 ,5 氯化三苯基四氮唑 (TTC)抑菌效价检测法 ,克隆形成法及动物移植性肿瘤模型测定Fab′ PYM偶联物在体外的生物学活性与在小鼠体内的抗肿瘤作用。结果　Fab′ PYM与BEL 740 2、肝癌 2 2 (H2 2 )及HT 2 9细胞呈免疫学阳性反应 ,但与KB细胞无反应 ;偶联物中PYM的抑菌活性为游离PYM的 15 % ;Fab′ PYM对体外培养的BEL 740 2细胞、HT 2 9细胞和KB细胞的 5 0 %抑制浓度 (IC50 )分别为 0 0 2、0 0 8及 0 5 0μmol/L ;静脉注射 10mg/kgFab′ PYM偶联物和PYM皮下接种对小鼠肝癌 2 2的抑瘤率分别为 86 %和6 2 % ;腹腔注射 ,10mg/kgFab′ PYM偶联物和PYM对小鼠脾内接种的结肠癌 2 6肝转移的抑制率分别为 91%和 6 2 %。结论　Fab′ PYM偶联物具有比游离平阳霉素更强的抑制肝癌生长与肿瘤肝转移作用。Objective To develop an immunoconjugate with inhibitory effect on the growth of hepatoma and hepatic metastasis of tumor by linking Fab′fragment of monoclona antibody (McAb) to pingyangmycin (PYM). Methods Monoclonal antibody Fab′ fragment was obtained by enzymolysis with pepsin and DTT reduction. Linking of the Fab′ fragment and PYM was mediated by dextran T40. Indirect ELISA, TTC bacteriostatic titer test, clonogenic assay and animal models transplanted with tumor were used to assess the biological activities and antitumor effects of Fab′PYM conjugate within and without the bodies of 10 mice. Results The Fab′PYM conjugate showed immunoreactivity to BEL7402, hepatoma 22 (H22) and HT29 cells, but no reaction to KB cells. The bacteriostatic activity of PYM in the conjugate was 15% as much as that of free PYM. The 50% inhibitory doses (IC50) of Fab′PYM conjugate to BEL7402, HT29 cells and KB cells were 0.02 μmol/L, 0.08 μmol/L and 0 50 μmol/L respectively. Given intravenously, 10 mg/kg × 6, Fab′PYM conjugate and PYM suppressed the growth of hepatoma 22 by 86% and 62% respectively. Given intraperitoneally, 10 mg/kg × 4, Fab′PYM and PYM decreased the hepatic metastasis of colon carcinoma 26 by 91% and 62% respectively. Conclusion Fab′PYM conjugate is more effective in combating hepatoma and hepatic metastasis of tumor than free PYM.

关 键 词：单克隆抗体 抗肿瘤抗生 平阳霉素 肝癌 抑制 肝转移瘤 

分 类 号：R735.7[医药卫生—肿瘤] R979.14[医药卫生—临床医学]