摘要
目的研究量子点标记靶向探针对人肝癌裸鼠模型的体内成像技术。方法将巯基乙酸修饰的水溶性量子点结合鼠抗人甲胎蛋白(AFP)单克隆抗体制备成水溶性量子点-AFP-Ab复合物探针。荧光、紫外光光谱分析及透射电镜研究其特性。通过直接免疫荧光法,用该复合物探针特异性识别肝癌细胞株HCCLM6 AFP抗原。将体外培养的肝癌细胞株HCCLM6通过皮下接种和尾静脉注射裸鼠分别建立人肝癌裸鼠模型和肺转移模型。尾静脉注射量子点-AFP-Ab探针,用蓝光二极管照射获得活体荧光成像;用掺Ti蓝宝石激光器照射,对肿瘤部位和正常部位进行光谱分析。取血清检测丙氨酸转氨酶、天冬氨酸转氨酶、尿素氮和肌苷水平。取裸鼠肝、脾、肾、肺、心和脑6种主要实质性器官行振荡切片,共聚焦显微镜观察,研究量子点-AFP-Ab探针在裸鼠体内的非特异性摄取.结果量子点-AFP-Ab复合物探针具有激发光谱宽、荧光强度高的特点,能特异性与肝癌细胞AFP抗原高亲和力结合,在体内能特异性靶向肿瘤组织进行活体成像,无明显急性毒性。光谱分析显示量子点- AFP-Ab复合物探针主要分布于肿瘤的外周部位,少数该探针被肝、脾和肺非特异性摄取。结论量子点-AFP-Ab复合物探针具有优良的光学特性和生物相容性,能够进行肝癌体内靶向成像,将有助于肝癌的分子靶向研究。
Objective To explore in-vivo targeted imaging techniques for liver cancer detection using quantum dots （QDs） labeled probes in a nude mouse model of human hepatocellular carcinoma. Methods Mercaptoacetic acid （MAA） modified QDs were linked to mouse-anti-human alpha-fetoprotein （AFP） monoclonal antibody to form water soluble QD-AFP-Ab probes, which were validated by spectra analyses and transmission electron microscope. The probes were firstly used to detect AFP antigen in human hepatocellular carcinoma cell line HCCLM6 in-vitro by one-step immunofluorescence method, ln-vivo tumor xenografts and lung metastases models were then established by inoculation of HCCLM6 cells subcutaneously and into the tail vein of nude mice, respectively. QD-AFP-Ab probes were injected into the tail vein of the tumor bearing mice for live animal fluorescence imaging. Spectra of tumor and normal tissue were analyzed under illumination of Ti： sapphire laser. Serum levels of alanine amino transferase, aspartate amino transferase, blood urea nitrogen and creatinine were determined by conventional biochemical analysis. The liver, spleen, lungs, kidneys, heart and brain of the experimental nude mice were investigated for nonspecific uptake of the probes by confocal microscope. Results The QD-AFP-Ab probes had broad excitation spectra and high fluorescence intensity. They could specifically and efficiently recognize AFP antigen in hepatocellular carcinoma cells. Tumor targeting imaging using these probes were successful without any acute toxicity to the experimental animals. Spectra analysis showed that the probes per field were lower in the centre than the periphery of the tumor. Non-specific uptake of QD-AFP-Ab probes occurred mainly in the liver, spleen and lungs. Conclusions QD-AFP-Ab probes have good optical properties and biocompatibility for in-vivo targeted imaging of hepatocellular carcinoma. Such approach promises to be highly desirable for molecular targeted research of liver cancer.
出处
《中华病理学杂志》
CAS
CSCD
北大核心
2007年第6期394-399,共6页
Chinese Journal of Pathology
基金
国家自然科学基金(20675058)
国家自然科学基金创新群体(20621502)
教育部新世纪优秀人才支持计划资金(NCET-04-0669)
全国优秀博士学位论文作者专项资金(200464)
关键词
癌
肝细胞
量子点
荧光抗体技术
Carcinoma,hepatocellular
Quantum dots
Fluorescent antibody technique
