99^Tc^m-抗ProGRP（31-98）单克隆抗体D-D3在荷小细胞肺癌裸鼠的体内生物分布及放射免疫显像研究  

Biodistribution and radioimmunoimaging of 99^Tc^m-labeled anti-ProGRP monoclonal antibody D-D3 in nude mice bearing SCLC xenografts

作　　者：谢亦驰[1] 周小林[4] 郝丽君[2] 洪智慧[1] 沈明敬[3] 徐忠恒[3] 石怡珍[1] 刘增礼[1] 

机构地区：[1]苏州大学附属第二医院核医学科,215004 [2]苏州大学附属第二医院超声科,215004 [3]苏州大学附属第二医院胸心外科,215004 [4]中国辐射防护研究院

出　　处：《中华核医学与分子影像杂志》2015年第5期359-363,共5页Chinese Journal of Nuclear Medicine and Molecular Imaging

基　　金：基金项目：卫生部核医学重点实验室及江苏省分子核医学重点实验室开放课题（KF201307）;苏州市“科教兴卫”青年科技项目（KJXW2012015）

摘　　要：目的研究99^Tc^m-标记的抗胃泌素释放肽前体[ProGRP（13-98）]单克隆抗体D-D，在荷人小细胞肺癌（SCLC）裸鼠体内的生物分布及放射免疫显像情况。方法（1）采用2-巯基乙醇还原法制备99^Tc^m-D-D3，凝胶柱分离法纯化标记产物，纸层析法检测标记率、放化纯、比活度及稳定性，细胞结合分析法测定标记抗体的抗原结合能力。（2）建立SCLCNCI-H446及肺腺癌A549荷瘤裸鼠模型。（3）自NCI-H446荷瘤裸鼠尾静脉注射99^Tc^m-D-D，后，于不同时间点处死裸鼠并取血液及各主要脏器组织标本，计算各时间点的放射性摄取值（％ID／g）和肿瘤／肌肉、肿瘤／血液的放射性计数比值。（4）对荷瘤裸鼠模型进行放射免疫显像，计算T／NT比值。采用SPSS17．0软件处理数据，两样本均数比较采用两样本t检验。结果（1）99^Tc^m-D-D3标记率为（73．87±2．89）％，放化纯为（94．13±4．49）％，比活度为（1．64-＋0．06）MBq!txg。与生理盐水及健康人血清充分混合，37℃放置4h后的放化纯分别为（82．17~1．oo）％和（74．13±1．67）％，12h后的放化纯分别为（55．734-1．07）％和（54．07±1．88）％。99^Tc^m-D-D3与NCI—H446和A549细胞结合率分别为（81．20±2．37）％和（24．30±1．46）％。（2）NCI—H446荷瘤裸鼠体内生物分布研究显示：注射99^Tc^m-D-D3后8h，瘤体的放射性摄取值[（2．13-＋0．06）％ID／g]已明显高于除肝、肾、胃及血液外的其他脏器及组织（t=2．52～24．53，均P〈0．05），12h时显著高于其他脏器及组织（t：1．98～20．88，均P〈0．05），24h时达最高值[（4．16＋0．45）％ID／g]；肿瘤／肌肉、肿瘤／血液的放射性比值随时间延长而逐渐升高。（3）放射免疫显像结果显示：注射后4h，NCI—H446荷瘤裸鼠的瘤体已较清晰，随时间延长移植瘤部位的放射性逐渐增多。A549荷瘤裸鼠注射99^Tc^m-D-D3后2Objective To explore the biodistribution and radioimmunoimaging of 99 ^Tc^m -labeled an- ti-ProGRP monoclonal antibody D-D3 in nude mice bearing human small cell lung cancer （SCLC） xenografts. Methods （1） D-D3 antibody was labeled with 99Tcm via the 2-mercaptoethanol reduction method. 99^Tc^m-D-D3 was purified by the gel column separation method. The labeling efficiency, radiochemical purity, specific ra- dioactivity and stability of 99^Tc^m-D-D3 were measured by thin-layer chromatography. Antigen-binding ability was determined by cell binding assay. （2）After tail vein injection of 99^Tc^m-D-D3, nude mice bearing SCLCNCI-H446 xenografts were sacrificed at designated time points. The blood and major organs were removed and weighed, and counted in a gamma scintillation counter. Radioactive uptake in each organ （% ID/g）, tumor/muscle and tumor/blood ratios at each time point were calculated. （3） After injection of 99^Tc^m-D-D3 , continuous imaging of the nude mice bearing NCI-H446 and lung adenoearcinoma A549 xenografts were car- ried out respectively, and the T/NT ratios were calculated. Two-sample t test was used to analyze the data. Results （ 1 ） The labeling rate of 99^Tc^m-D-D3 was （73.87＋2.89）%. The radiochemical purity was （94.13±4. 49） %. The specific radioactivity was （ 1.64 ± 0.06 ） MBq/μg. Being fully mixed with either saline or healthy human serum and incubated at 37 ℃ for 4 h, the radiochemical purities of 99Tern-D-D3 were （82.17±1.00） % and （ 74.13± 1.67 ） % respectively, and （ 55.73±1.07 ） % and （ 54.07±1.88 ） % respectively after being incubated for 12 h. The immunobinding rates of 99^Tc^m-D-D3 to NCI-H446 and A549 cells were （81.20±2.37）% and （24.30±1.46）%, respectively. （2）The %ID/g of 99^Tc^m-D-D3 in NCI-H446 tumor was 2.13-＋ 0.06, which was significantly higher than those of other organs except liver, kidneys, stomach and blood at 8 h after injection （t = 2.52-24.53, all P〈0.05）. The uptake in the tumors was high

关 键 词：癌 小细胞 肺 抗体 单克隆 胃泌素释放肽 放射性核素显像 小鼠 裸 

分 类 号：R734.2[医药卫生—肿瘤] R730.44[医药卫生—临床医学]