^(89)Zr标记EGFR及HER2抗体用于胃黏液腺癌的microPET显像研究  

MicroPET imaging studies of^(89)Zr-labeled EGFR and HER2 antibodies in gastric mucinous adenocarcinoma

作　　者：石岱 曹庆辉 张颖颖 程远 徐展 程登峰 石洪成 Shi Dai;Cao Qinghui;Zhang Yingying;Cheng Yuan;Xu Zhan;Cheng Dengfeng;Shi Hongcheng(Shanghai Institute of Medical Imaging Department of Nuclear Medicine,Zhongshan Hospital,Fudan University Nuclear Medicine Institute of Fudan University Cancer Center,Zhongshan Hospital,Fudan University,Shanghai 200032,China;Department of Pharmacy,Zhongshan Hospital,Fudan University,Shanghai 200032,China)

机构地区：[1]上海市影像医学研究所、复旦大学附属中山医院核医学科、复旦大学核医学研究所、复旦中山医院肿瘤中心,200032 [2]复旦大学附属中山医院药剂科,上海200032

出　　处：《中华核医学与分子影像杂志》2021年第10期597-601,共5页Chinese Journal of Nuclear Medicine and Molecular Imaging

基　　金：复旦大学附属中山医院临床研究专项基金(2020ZSLC20)。

摘　　要：目的筛选适合监测低葡萄糖代谢的胃黏液腺癌小鼠模型的^(89)Zr标记表皮生长因子受体(EGFR)和人表皮生长因子受体2(HER2)单克隆抗体(简称单抗)分子探针。方法通过细胞爬片和移植瘤肿瘤切片验证MGC803胃癌细胞株的EGFR和HER2表达情况;用^(89)Zr标记去铁胺-西妥昔单抗(DFO-Cetuximab)和去铁胺-帕妥珠单抗(DFO-Pertuzumab),制得分别靶向EGFR和HER2的^(89)Zr-DFO-Cetuximab和^(89)Zr-DFO-Pertuzumab,测定其放化纯;通过细胞结合实验、阻断实验验证^(89)Zr-DFO-Cetuximab和^(89)Zr-DFO-Pertuzumab与MGC803的结合力和特异性;将12只MGC803荷瘤裸鼠模型分3组(每组4只),分别注射^(89)Zr-DFO-Cetuximab(7.4 MBq/只,74μg/只)、^(89)Zr-DFO-Pertuzumab(7.4 MBq/只,70μg/只)和18F-脱氧葡萄糖(FDG)(7.4 MBq/只),于注射后4、24和48 h进行microPET显像(18F-FDG显像为注射后1 h);另取8只荷瘤裸鼠,分为^(89)Zr-DFO-Cetuximab组和^(89)Zr-DFO-Pertuzumab组(各4只),于探针注射后48 h进行生物分布研究。采用两独立样本t检验进行组间生物分布比较。结果肿瘤切片免疫荧光染色示MGC803胃癌细胞株EGFR表达量高于HER2。^(89)Zr-DFO-Cetuximab和^(89)Zr-DFO-Pertuzumab放化纯均大于95%,比活度分别为100和95 MBq/mg;2种探针在生理盐水和胎牛血清(FBS)中稳定性好,放置72 h放化纯仍高于80%。MicroPET显像示MGC803肿瘤部位^(89)Zr-DFO-Cetuximab的摄取高于18F-FDG和^(89)Zr-DFO-Pertuzumab。生物分布实验示,48 h肿瘤^(89)Zr-DFO-Cetuximab摄取[每克组织百分注射剂量率(%ID/g)]为56.3±12.0,高于^(89)Zr-DFO-Pertuzumab摄取(22.0±3.6;t=4.31,P<0.05)。结论相较于^(89)Zr-DFO-Pertuzumab,^(89)Zr-DFO-Cetuximab具有更好的无创监测低葡萄糖代谢胃黏液腺癌的潜能。Objective To screen ^(89)Zr-labeled anti-epidermal growth factor receptor(EGFR)and human epidermal growth factor receptor 2(HER2)monoclonal antibody molecular probes suitable for monitoring the gastric mucinous adenocarcinoma bearing mouse models with low glucose metabolism.Methods The expression of EGFR and HER2 in the MGC803 gastric cancer cell line was verified by analyzing cell slides and xenograft tumor sections.^(89)Zr-Deferoxamine(DFO)-Cetuximab and ^(89)Zr-DFO-Pertuzumab were prepared and the radiochemical purity was detected.Cell binding experiments and blocking experiments were performed to verify the binding ability and specificity of the probes.Twelve gastric mucinous adenocarcinoma bearing mouse models were divided into 3 groups(n=4 in each group):^(89)Zr-DFO-Cetuximab group(7.4 MBq/mouse,74μg/mouse),^(89)Zr-DFO-Pertuzumab group(7.4 MBq/mouse,70μg/mouse)and 18F-fluorodeoxyglucose(FDG)group(7.4 MBq/mouse).MicroPET imaging was performed at 4,24 and 48 h(18F-FDG group underwent imaging at 1 h only)post-injection.The biodistribution study of ^(89)Zr-DFO-Cetuximab and ^(89)Zr-DFO-Pertuzumab was conducted in 2 groups(n=4 in each group)48 h after the injection.The independent sample t test was used for data analysis.Results The immunofluorescent staining demonstrated EGFR expression was significantly higher than HER2 expression in MGC803 gastric cancer cell line.The radiochemical purity of ^(89)Zr-DFO-Cetuximab and ^(89)Zr-DFO-Pertuzumab were both more than 95%,and the specific activities were 100 and 95 MBq/mg,respectively.The two probes had good stability in normal saline and fetal bovine serum,with the radiochemical purity higher than 80%at 72 h.MicroPET imaging showed that the uptake of ^(89)Zr-DFO-Cetuximab in the MGC803 tumor was significantly higher than that of 18F-FDG and ^(89)Zr-DFO-Pertuzumab.The biodistribution study demonstrated the ^(89)Zr-DFO-Cetuximab uptake(percentage activity of injection dose per gram of tissue,%ID/g)of tumors at 48 h was significantly higher than that of ^(89)Zr-DFO-Pe

关 键 词：腺癌 黏液 胃 受体 表皮生长因子 同位素标记 锆 正电子发射断层显像术 肿瘤细胞 培养的 小鼠 裸 

分 类 号：R735.2[医药卫生—肿瘤] R730.44[医药卫生—临床医学]