Survivin反义寡核苷酸的^(99m)Tc标记及在肝癌细胞中的表达  

Labeling of survivin antisense oligonucleotide with ^(99m)Tc and the research on the expression of ^(99m) Tc surviving ASON in hepatocellular carcinoma

作　　者：高再荣[1] 张凯军[1] 张永学[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院核医学科,武汉430022

出　　处：《放射学实践》2004年第1期50-52,共3页Radiologic Practice

基　　金：湖北省科技厅自然科学基金资助项目 (编号 2 0 0 2AB1 2 1 )

摘　　要：目的 :研究放射性核素99mTc标记Survivin反义寡核苷酸 (ASON)的方法及在肝癌细胞中的动力学 ,探讨用于肝癌诊断的可能性。方法 :用DNA合成仪合成 18碱基单链SurvivinASON ,在 5′末端经氨基修饰后 ,用S acetyl NHS MAG3作螯合剂 ,进行99mTc标记SurvivinASON ,并对其标记率、放化纯、比活度、稳定性、细胞摄取率和细胞清除率进行了研究。结果 :99mTc MAG3 ASON的标记率为 65 .2 2 %,放化纯为 94%,比活度为 1190kBq/μg。室温条件下与生理盐水温育 4h后 ,其放化纯为 92 %。 3 7℃ 5 %CO2 条件下与新鲜人血清温育 4h ,未见明显的血清蛋白结合作用。肝癌细胞摄取率 10h达 12 .73 %,细胞滞留率 8h为 5 4.2 2 %。结论 :采用S acetyl NHS MAG3 作螯合剂标记Survivin ASON的方法可行 ,可获得较好的标记效果 ,99mTc survivin ASON在肝癌细胞中有较理想的摄取率 。Objective:To establish a new labeling method of survivin antisense oligonucleotide (ASON) with technetium 99m ( 99m Tc) and investigate the possibility of diagnosing hepatocellular carcinoma using 99m Tc surviving ASON labeling.Methods:An 18 base single stranded antisense oligonucleotide sequence was synthesized and linked with a primary amine on the 5′end.The survivin ASON was labeled with 99m Tc by conjugating the chelator Sacetyl NHS MAG 3.The properties of 99m Tc surviving ASON,such as radiolabeling efficiency,radiochemical purity,were studied.Results:The radiolabeling efficiency, radiochemical purity and specific activity of 99m Tc surviving ASON were 65.22%,94% and 1190kBq/μg, respectively.The accumulation rate in hepatocellular carcinoma reached 12.73% at 10h.After incubation with normal saline at room temperature for 4h,the radiochemical purity was still 92%.No evidence for serum protein binding was observed after incubation with fresh human serum at 37℃ in 5% CO 2 for 4h.Conclusion: The labeling method of survivin ASON with 99m Tc,conjugated by Sacetyl NHS MAG 3,has been successfully proved. 99m Tc surviving ASON can be effectively taken in hepatocellular carcinoma,which may be used hopefully for clinical diagnosis of hepatocellular carcinoma in the future.

关 键 词：SURVIVIN基因 反义寡核苷酸 ^99MTC 肝癌细胞 基因显像诊断 肿瘤标志物 

分 类 号：R735.7[医药卫生—肿瘤]