Lin Jiaying, Hu Weiwei, Gao Fengli, Qin Jinbao, Peng Chen, Lu Xinwu
Department of Assisted Reproduction, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University, School of Medicine, 639 Zhizaoju Road, Shanghai 200011, China.
Department of Critical Care Medicine, the Affiliated Hospital of Xuzhou Medical University, Jiangsu, 2210021, PR China.
J Cancer. 2018 Jan 1;9(3):564-577. doi: 10.7150/jca.19786. eCollection 2018.
It has been a great challenge to develop multifunctional fluorescent nanoprobes for tumor-targeted imaging. In this study, we developed folic acid (FA) gold nanoparticles (AuNPs) through diatrozic acid (DTA) linking for and targeted imaging of HeLa cervical cancer cells by computed tomography (CT). G5 dendrimers were used as templates to synthesize AuNPs within the interiors of dendrimers. The synthesized AuNPs were then sequentially modified by fluorescein isothiocyanate, FA, and DTA and the remaining terminal amines on the dendrimers were acetylated. We further performed hematoxylin and eosin staining, cell viability assay, flow cytometric analysis of cell cycle and apoptosis, and hemolytic assay to examine the cytotoxicity and hemocompatibility of the particles. The specific uptake of the nanoparticles by HeLa cells was determined through inductively coupled plasma atomic emission spectroscopy determination of silver and transmission electron microscopy. Lastly, HeLa cells and a xenografted HeLa tumor model were employed to evaluate the and targeted CT imaging performances of the nanoparticles, respectively. We showed that Au DENPs-FA-DTA does not cause cytotoxic effects on both HeLa cancer cells and healthy normal cells in mice, demonstrating the superior biocompatibility and stability of the particles in the given concentration range. Micro-CT images documented that HeLa cells incubated with Au DENPs-FA-DTA could be identified by X-ray examinations and that HeLa cells xenografts in BALB/c nude mice could be imaged after the mice were administered with the particles intravenously or intratumorally. The FA-modified AuNPs enabled targeted CT imaging of HeLa cells overexpressing FA receptors and . Taken together, our results showed that the AuNPs we developed exhibit great potentials as imaging probes for targeted CT imaging of human cervical cancer.
开发用于肿瘤靶向成像的多功能荧光纳米探针一直是一项巨大的挑战。在本研究中,我们通过重氮酸(DTA)连接开发了叶酸(FA)金纳米颗粒(AuNPs),并通过计算机断层扫描(CT)对HeLa宫颈癌细胞进行靶向成像。以G5树枝状大分子为模板在树枝状大分子内部合成AuNPs。然后将合成的AuNPs依次用异硫氰酸荧光素、FA和DTA进行修饰,并将树枝状大分子上剩余的末端胺乙酰化。我们进一步进行了苏木精和伊红染色、细胞活力测定、细胞周期和凋亡的流式细胞术分析以及溶血试验,以检测颗粒的细胞毒性和血液相容性。通过电感耦合等离子体原子发射光谱法测定银和透射电子显微镜确定HeLa细胞对纳米颗粒的特异性摄取。最后,分别使用HeLa细胞和异种移植的HeLa肿瘤模型评估纳米颗粒的靶向CT成像性能。我们表明,Au DENPs-FA-DTA对小鼠的HeLa癌细胞和健康正常细胞均无细胞毒性作用,证明了颗粒在给定浓度范围内具有优异的生物相容性和稳定性。显微CT图像记录显示,用Au DENPs-FA-DTA孵育的HeLa细胞可通过X射线检查识别,并且在给BALB/c裸鼠静脉内或瘤内注射颗粒后,异种移植的HeLa细胞可以成像。FA修饰的AuNPs能够对过表达FA受体 和 的HeLa细胞进行靶向CT成像。综上所述,我们的结果表明,我们开发的AuNPs作为人宫颈癌靶向CT成像的成像探针具有巨大潜力。