Chen Liang-dong, Liu Jia, Yu Xue-feng, Pang Dai-wen, Wang Qu-quan, Yuan Hong-yin, Tang Zhao-you, Li Yan
Zhongnan Hospital of Wuhan University and Cancer Center of Wuhan University, Wuhan 430071, China.
Zhonghua Bing Li Xue Za Zhi. 2007 Jun;36(6):394-9.
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.
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. In-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.
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.
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.
在人肝细胞癌裸鼠模型中，探索使用量子点（QD）标记探针进行肝癌检测的体内靶向成像技术。
将巯基乙酸（MAA）修饰的量子点与鼠抗人甲胎蛋白（AFP）单克隆抗体连接，形成水溶性QD-AFP-Ab探针，通过光谱分析和透射电子显微镜进行验证。首先采用一步免疫荧光法，在体外检测人肝癌细胞系HCCLM6中的AFP抗原。然后分别通过将HCCLM6细胞皮下接种和经尾静脉接种到裸鼠体内，建立体内肿瘤异种移植和肺转移模型。将QD-AFP-Ab探针经尾静脉注射到荷瘤小鼠体内进行活体动物荧光成像。在钛宝石激光照射下分析肿瘤和正常组织的光谱。通过常规生化分析测定血清谷丙转氨酶、谷草转氨酶、血尿素氮和肌酐水平。利用共聚焦显微镜研究实验裸鼠的肝脏、脾脏、肺脏、肾脏、心脏和大脑对探针的非特异性摄取情况。
QD-AFP-Ab探针具有宽激发光谱和高荧光强度。它们能够特异性且高效地识别肝癌细胞中的AFP抗原。使用这些探针进行肿瘤靶向成像取得成功，且对实验动物无任何急性毒性。光谱分析表明，每个视野中肿瘤中心的探针含量低于周边。QD-AFP-Ab探针的非特异性摄取主要发生在肝脏、脾脏和肺脏。
QD-AFP-Ab探针在肝癌体内靶向成像方面具有良好的光学特性和生物相容性。这种方法有望成为肝癌分子靶向研究的理想手段。
