Wang Chung-Hao, Huang Yao-Jhang, Chang Chia-Wei, Hsu Wen-Ming, Peng Ching-An
Department of Chemical Engineering, National Taiwan University, Taipei, Taiwan.
Nanotechnology. 2009 Aug 5;20(31):315101. doi: 10.1088/0957-4484/20/31/315101. Epub 2009 Jul 13.
Despite aggressive multimodality therapy, most neuroblastoma-bearing patients relapse and survival rate remains poor. Exploration of alternative therapeutic modalities is needed. Carbon nanotubes (CNTs), revealing optical absorbance in the near-infrared region, warrant their merits in photothermal therapy. In order to specifically target disialoganglioside (GD2) overexpressed on the surface of neuroblastoma stNB-V1 cells, GD2 monoclonal antibody (anti-GD2) was conjugated to acidified CNTs. To examine the fate of anti-GD2 bound CNTs after incubation with stNB-V1 cells, rhodamine B was labeled on carboxylated CNTs functionalized with and without anti-GD2. Our results illustrated that anti-GD2-linked CNTs were extensively internalized by neuroblastoma cells via GD2-mediated endocytosis. In addition, we showed that anti-GD2 bound CNTs were not ingested by PC12 cells without GD2 expression. After anti-GD2 conjugated CNTs were incubated with neuroblastoma cells for 6 h and endocytosed by the cells, CNT-laden neuroblastoma cells were further irradiated with an 808 nm near-infrared (NIR) laser with intensity ramping from 0.6 to 6 W cm(-2) for 10 min which was then maintained at 6 W cm(-2) for an additional 5 min. Post-NIR laser exposure, and after being examined by calcein-AM dye, stNB-V1 cells were all found to undergo necrosis, while non-GD2 expressing PC12 cells all remained viable. Based on the in vitro study, CNTs bound with anti-GD2 have the potential to be utilized as a therapeutic thermal coupling agent that generates heat sufficient to selectively kill neuroblastoma cells under NIR laser light exposure.
尽管采用了积极的多模态治疗,但大多数患有神经母细胞瘤的患者仍会复发,生存率仍然很低。因此需要探索其他治疗方式。碳纳米管(CNTs)在近红外区域具有光吸收特性,这使其在光热治疗中具有优势。为了特异性靶向神经母细胞瘤stNB-V1细胞表面过度表达的二唾液酸神经节苷脂(GD2),将GD2单克隆抗体(抗GD2)与酸化的碳纳米管偶联。为了检测抗GD2结合的碳纳米管与stNB-V1细胞孵育后的命运,将罗丹明B标记在未偶联和偶联抗GD2的羧基化碳纳米管上。我们的结果表明,抗GD2连接的碳纳米管通过GD2介导的内吞作用被神经母细胞瘤细胞广泛内化。此外,我们还表明,没有GD2表达的PC12细胞不会摄取抗GD2结合的碳纳米管。抗GD2偶联的碳纳米管与神经母细胞瘤细胞孵育6小时并被细胞内吞后,用强度从0.6增加到6 W cm(-2)的808 nm近红外(NIR)激光照射负载碳纳米管的神经母细胞瘤细胞10分钟,然后保持在6 W cm(-2)再照射5分钟。近红外激光照射后,用钙黄绿素-AM染料检测发现,stNB-V1细胞全部发生坏死,而不表达GD2的PC12细胞全部存活。基于体外研究,与抗GD2结合的碳纳米管有潜力用作治疗性热偶联剂,在近红外激光照射下产生足以选择性杀死神经母细胞瘤细胞的热量。