抗肝癌单链抗体免疫脂质体的制备及其体外抑瘤实验  

Preparation of anti-hepatocellular carcinoma scFv immunoliposome and its killing effects on hepatocarcinoma cells in vitro

作　　者：张桂红[1] 刘彦仿[1] 付勇[1] 胡海洋[2] 陈大为[2] 杨守京[1] 赵君[1] 

机构地区：[1]第四军医大学病理学教研室,陕西西安710032 [2]沈阳药科大学药学院,辽宁沈阳110016

出　　处：《医学研究生学报》2006年第1期3-5,共3页Journal of Medical Postgraduates

基　　金：国家自然科学基金资助项目(批准号:30370810)

摘　　要：目的:比较抗肝癌单链抗体免疫脂质体(hdsFv25-lip.PE38,简称免疫脂质体PE38)、抗肝癌单链抗体免疫毒素(hdsFv-PE38,简称免疫毒素PE38)和载PE38脂质体(lip.PE38,简称脂质体PE38)对体外培养的肝癌细胞SMMC-7721的杀伤作用。方法:将磷脂(PC)、胆固醇(CHOL)和胆固醇琥珀酰酯(CHS)按一定比例溶于氯仿。除去氯仿后形成干膜,加入PE38蛋白溶液,超声、过滤及过Sephadex G-50柱子。测包峰率后加入乙基[3-(二甲胺基)丙基]、碳二亚胺盐酸盐(EDC)和氮羟基琥珀酰亚胺碘酸钠(SSNHS)在2-(n-吗啡啉)乙磺酸(MES)缓冲液中反应30m in,再加单链抗体(hdscFv),4℃过夜。用单四唑(MTT)法检测所制备的免疫脂质体、免疫毒素和脂质体对肝癌细胞的杀伤作用。结果:通过MTT法显示:免疫脂质体对肝癌细胞的杀伤效果最好,免疫毒素次之,脂质体最差,其半抑制浓度(IC50)值分别为0.59μg/m l、6.04μg/m l和92.07μg/m l。结论:载PE38免疫脂质体有望成为良好的新型抗肝癌药物。Objective： To compare the killing effects on HCC SMMC-7721 cells of immunoliposomePE38 ,immunotoxin PE38 and liposome PE38. Methods：Dissolute PC, CHOL and CHS in chloroform in proportion. Dry membrane was formed after chloroform was removed. Add the protein solution of PE38 to dissolute the dry membrane. Then pass the solution over a Sephadex G-50 column after uhrasoned and filtrated to detect the encapsulation efficiency of the liposome. The solution reacted in EDC, SSNHS and MES for 30 minutes. Then add Ab to the solution in 4℃ over night. MTI method was used to detect the killing effects on HCC cells of immunoliposome PE38 ,immunotoxin PE38 and liposome PE38 in vitro. Results ：The killing effects on HCC cells of immunoliposome PE38 is the best, and that of liposome PE38 is the worst. The respective IC50 s are ：0.59 μg/m1,6.04 μg/ml and 92.07 μg/ml. Conclusion ; Immunoliposome PE38 may be a protential durg in the treatment of hepatocarinoma.

关 键 词：免疫脂质体 脂质体 肝肿瘤/病理学 比色法 

分 类 号：R735.7[医药卫生—肿瘤]