摘要
目的:研究放射性核素碘(131I)标记表皮生长因子受体单克隆抗体(McAb)1H12的实验条件,观察标记产物131I-1H12与人肺癌细胞A549的免疫结合及生物学效应。方法:131I以Iodogen法标记1H12,经体外细胞结合试验分析检测标记抗体的免疫活性;通过流式细胞仪检测不同剂量131I-1H12(148、74、37kBq)、游离131I(148kBq)及1H12(80nmol·ml-1)对A549细胞的作用。结果:131I-1H12标记率为50.14%,比活度为4.63MBq·μg-1,放射性浓度为77.31MBq·ml-1,放射性化学纯度为96.92%。131I-1H12与A549细胞结合率为61.12%;131I-1H12诱导A549细胞凋亡和周期阻滞呈剂量-效应关系,以148kBq131I-1H12作用效应最大,凋亡率达到(44.35±3.31)%,G2/M期细胞阻滞率达(51.17±2.98)%。结论:131I-1H12能够与A549细胞发生免疫结合,并调控细胞周期和诱导细胞凋亡。
Objective：To investigate the experimental conditions of radionuclide iodine（131I） labeling epidermal growth factor receptor（EGFR） monoclonal antibody（McAb） 1H12,observe the cell-binding function and biological effects of 131I-1H12 to human lung cancer cells A549.Methods：1H12 was labeled with 131I by Iodogen method.The immunocompetence of 131I-1H12 was analyzed by cell-binding test.The cell apoptosis and cell cycles of A549 were analyzed by flow cytometry assay with various doses of 131I-1H12（148,74,37 kBq）,free131 I（148 kBq）,1H12（80 nmol·ml-1） and equivalent medium.Results：The labeling rate of 131I-1H12 was 50.14%,its specific activity,radioactive concentration and radiochemical purity were 4.63 MBq·μg-1,77.31 MBq·ml-1 and 96.92% respectively.The specific binding rate of 131I-1H12 to A549 cells was 61.12% ;The apoptosis rate and cycle blockage rate of A549 cells induced by 131I-1H12 were in a dose-effect relationship;The supreme apoptosis rate[（44.35 ±3.31） %]and G2 /M cell cycle blockage rate[（51.17 ±2.98） %]were found in the A549 cells treated with 148 kBq 131I-1H12.Conclusion：131I-1H12 can immunobind with A549 cells and regulate cell cycle and induce apoptosis of A549 cells to inhibit its proliferation;It might have some potential application prospect in biological targeted diagnosis and therapeutics.
出处
《东南大学学报（医学版）》
CAS
2010年第4期430-434,共5页
Journal of Southeast University(Medical Science Edition)
基金
江苏省重点技术创新项目(7625003030)
东南大学国家自然科学基金预研项目(9290001412)
