近红外荧光成像监测ENO1 mAb在宫颈癌荷瘤小鼠体内生物分布的研究  

Study on the Biological Distribution of ENO1 mAb in Cervical CancerBearing Mice Monitored by Near Infrared Fluorescence Imaging

作　　者：赵调红 章婷婷[1,2] 陈圆圆 路如霞 赵源旭 裴亚萍 刘会玲 祝秉东[3] ZHAO Tiaohong;ZHANG Tingting;CHEN Yuanyuan;LU Ruxia;ZHAO Yuanxu;PEI Yaping;LIU Huiling;ZHU Bingdong(The First Clinical Medical College,Gansu University of Traditional Chinese Medicine,Lanzhou 730000,Gansu,P.R.China;Department of Gynecology,Gansu Provincial Hospital,Lanzhou 730000,Gansu,P.R.China;School of Basic Medical Sciences,Lanzhou University,Lanzhou 730000,Gansu,P.R.China)

机构地区：[1]甘肃中医药大学第一临床医学院,甘肃兰州730000 [2]甘肃省人民医院妇科,甘肃兰州730000 [3]兰州大学基础医学院,甘肃兰州730000

出　　处：《影像科学与光化学》2023年第6期350-357,共8页Imaging Science and Photochemistry

基　　金：国家自然科学基金地区基金(82260557),北京科创医学发展基金会肿瘤研究基金面上项目(KC2021-JX-0186-141)。

摘　　要：本研究探讨近红外荧光成像(NIRF)观察烯醇化酶1(ENO1)单克隆抗体(mAb)在宫颈癌荷瘤小鼠体内的分布、代谢及肿瘤靶向效率。利用癌症基因组图谱分析ENO1的表达与肿瘤分期和预后以及肿瘤免疫浸润的关系;Cy7-NHS和FITC与ENO1 mAb偶联成Cy7-ENO1 mAb和FITC-ENO1 mAb,通过激光共聚焦显微镜验证ENO1 mAb在宫颈癌细胞TC-1的定位,并建立宫颈癌皮下肿瘤模型,尾静脉注射Cy7-ENO1 mAb后在1、6、24、48、72和120 h进行近红外荧光成像,成像24 h后迅速解剖主要脏器进行体外成像。生物信息学分析结果表明ENO1在正常宫颈组织与宫颈癌组织中的表达有显著差异,免疫组织化学(IHC)结果提示ENO1蛋白在宫颈癌组织中高表达;此外,其表达与肿瘤分期和淋巴结分期显著相关,并且ENO1的高表达与多种糖酵解酶相关;激光共聚焦显微镜扫描显示FITC-ENO1 mAb能特异性结合在TC-1细胞膜表面,细胞未见内化;近红外荧光成像证明Cy7-ENO1 mAb主要富集在肝脏、肾脏和肿瘤组织,并且具有较高的肿瘤靶向效率。NIRF成像可以实时、动态监测ENO1 mAb在宫颈癌荷瘤小鼠体内的分布,其代谢器官主要是肝脏和肾脏,并且证实ENO1 mAb具有靶向宫颈癌作用。In this study,the distribution,metabolism and tumor targeting efficiency of Enolase 1(ENO1)monoclonal antibody(mAb)in cervical cancer bearing mice were investigated by near infrared fluorescence imaging.The relationship between ENO1 expression and tumor stage, prognosis and tumor immune invasion was analyzed by cancer genome atlas. Cy7-NHS and FITC were coupled with ENO1 mAb to form Cy7-ENO1 mAb and FITC-ENO1 mAb, and the localization of ENO1 mAb in cervical cancer cell TC-1 was verified by laser confocal microscopy, and the subcutaneous tumor model of cervical cancer was established. Near infrared fluorescence (NIRF) imaging was performed at 1, 6, 24, 48, 72, and 120 h after caudal intravenous injection of Cy7-ENO1 mAb. After 24 h of imaging, major organs were rapidly dissected for in vitro imaging. Bioinformatics results showed significant differences in the expression of ENO1 between normal cervical tissue and cervical cancer tissue while immunohistochemistry (IHC) indicated higher ENO1 protein expression in cervical cancer tissues. In addition, the expression of ENO1 was significantly correlated with tumor stage and lymph node stage, and the high expression of ENO1 was correlated with multiple glycolytic enzymes. Confocal laser microscopy showed that FITC-ENO1 mAb could specifically bind to the surface of TC-1 cell membrane, and no cell internalization was observed. Near infrared fluorescence imaging showed that Cy7-ENO1 mAb was mainly enriched in liver, kidney and tumor tissues, and had high tumor targeting efficiency. NIRF imaging allows real-time and dynamic monitoring of ENO1 mAb distribution in cervical cancer bearing mice, whose metabolic organs are mainly liver and kidney, and confirms ENO1 mAb's targeting of cervical cancer.

关 键 词：近红外荧光成像 ENO1 mAb 生物分布 Cy7-NHS 

分 类 号：R737.33[医药卫生—肿瘤]