眼镜蛇膜毒素与单克隆抗体偶联物对鼻咽癌细胞的特异性杀伤作用  

Specific Killing of Human CNE2-Cell by a Monoclonal Antibody Coupled to Membrane Toxin from Naja Naja Atra

作　　者：李钦章[1] 黄沿中 杨冠群[1] 

机构地区：[1]暨南大学

出　　处：《暨南大学学报（自然科学与医学版）》1998年第3期94-99,共6页Journal of Jinan University(Natural Science & Medicine Edition)

基　　金：广东省中医药局科研基金

摘　　要：应用柱层析和凝胶过滤的方法，从中华眼镜蛇的粗毒中分离纯化出一个不含磷脂酶Ａ２（ＰＬＡ２）的膜毒素（Ｍｅｍｂｒａｎｅｔｏｘｉｎ，ＭＴ）组分．ＭＴ经电泳鉴定为单一成分．将ｑｄＭＴ与鼠抗人鼻咽癌细胞（ＣＮＥ２）的单克隆抗体（ＢＡＣ５）偶联制备成免疫毒素（ＢＡＣ５－ＭＴ），经间接免疫荧光检测，证实免疫毒素仍保留了很好的生物活性．体外抗癌实验证明，在无Ｃａ２＋培养基中，游离的ＭＴ对ＣＮＥ２和Ｓ１８０细胞均有快速杀伤作用，并且对２种细胞的杀伤作用没有明显的区别，但ＢＡＣ５－ＭＴ同样在无Ｃａ２＋条件下，仅对靶细胞ＣＮＥ２有强烈的杀伤作用（杀伤率达７８．６％），对Ｓ１８０细胞只有很弱的杀伤作用（杀伤率为２１．３％）．结果表明ＢＡＣ５－ＭＴ对ＣＮＥ２细胞有很好的选择性杀伤作用，提示蛇膜毒素在肿瘤的导向治疗中将是一条很有希望的途径．Membrane toxin (MT) without contamination of phospholipase A2 was isolated from Naja naja atra venom by chromatography on CM-Sephadex C-25 and DEAE-Sephadex A-50.The immunotoxin (IT)BAC 5-MT was prepared by conjugating MT with monoclonal antibody(McAb) BAC 5 directed to human CNE 2 cells.In vitro,free MT showed high cytotoxicity for CNE 2 and S180cells without Ca2+ but there was no great difference between them.However,under the same condition,there was agreat difference in the killing rate of IT for the CNE2 and S180 cells (786% of the CNE2 cells were killed at a concentration of 1.0×10-6mol·L-1,but only 21.3% of S180 cells were killed).The results suggest that BAC5-MT has potential application in tumor the erapy.Calcium can inhibit the cytolytic activity of MT.

关 键 词：膜毒素 单克隆抗体 鼻咽癌 癌细胞杀伤 眼镜蛇毒 

分 类 号：R739.630.5[医药卫生—肿瘤] R979.1[医药卫生—临床医学]