Mohd-Zahid Manali Haniti, Zulkifli Siti Nadiah, Che Abdullah Che Azurahanim, Lim JitKang, Fakurazi Sharida, Wong Kah Keng, Zakaria Andee Dzulkarnaen, Ismail Norzila, Uskoković Vuk, Mohamud Rohimah, Z A Iskandar
Department of Chemical Pathology, School of Medical Sciences, Universiti Sains Malaysia 16150 Kubang Kerian Kelantan Malaysia
Material Synthesis and Characterization Laboratory, Institute of Advanced Technology, Universiti Putra Malaysia Serdang 43400 Selangor Malaysia.
RSC Adv. 2021 Apr 30;11(26):16131-16141. doi: 10.1039/d1ra01093j. eCollection 2021 Apr 26.
The enhanced permeability and retention effect allows for passive targeting of solid tumours by nanoparticles carrying anticancer drugs. However, active targeting by incorporation of various ligands onto nanoparticles can provide for a more selective and enhanced chemotherapeutic effect and complement the deficiencies of the passive targeting approach. Here we report on the design of the carboxyl-terminated PEGylated gold nanoparticles (AuNPs), their functionalization with anti-CD133 monoclonal antibody (mAb) a crosslinking reaction, and subsequent 5-fluorouracil (5-FU) drug loading. The synthesized products in the form of stable colloids were characterised using a range of physicochemical techniques, including X-ray diffraction (XRD), UV-Vis spectroscopy, transmission electron microscopy (TEM), and dynamic light scattering (DLS). Conjugation of anti-CD133 mAb onto PEGylated AuNPs was confirmed with the use of UV-Vis, BCA protein assay and fluorescence microscopy. HCT116 colorectal cancer cells abundantly expressed CD133: 92.4 ± 1.3%, as measured by flow cytometry. Whereas PEGylated AuNPs not conjugated with anti-CD133 mAb accumulated mainly at the cellular membrane, nanoparticles conjugated with anti-CD133 mAb were contained within the nuclear region of the cells. Anti-CD133 mAb conjugation facilitated the specific intracellular uptake due to specific antigen-antibody binding interaction. cytotoxicity studies on HCT116 cells showed that PEGylated AuNPs and PEGylated AuNPs-CD133 did not elicit any toxicity at any of the tested concentrations. Meanwhile, 5-FU-PEGylated AuNPs-CD133 significantly reduced the cell viability relative to the treatment with 5-FU-PEGylated AuNPs without anti-CD133 mAb conjugates ( < 0.0001). This study shows that the conjugation of nanocarriers with the anti-CD133 antibody improves the specific targeting of 5-FU against colorectal cancer cells. These results demonstrate that simultaneous functionalisation of PEGylated AuNPs with antibodies and chemotherapeutic drugs is a viable strategy to combat cancer through targeted drug delivery.
增强的渗透与滞留效应使得携带抗癌药物的纳米颗粒能够被动靶向实体瘤。然而，通过在纳米颗粒上结合各种配体进行主动靶向，可以提供更具选择性且增强的化疗效果，并弥补被动靶向方法的不足。在此，我们报告了羧基末端聚乙二醇化金纳米颗粒（AuNPs）的设计、其与抗CD133单克隆抗体（mAb）的功能化、交联反应以及随后的5-氟尿嘧啶（5-FU）药物负载。使用一系列物理化学技术对以稳定胶体形式合成的产物进行了表征，包括X射线衍射（XRD）、紫外可见光谱、透射电子显微镜（TEM）和动态光散射（DLS）。通过紫外可见光谱、BCA蛋白测定和荧光显微镜证实了抗CD133 mAb与聚乙二醇化AuNPs的结合。通过流式细胞术测定，HCT116结肠癌细胞大量表达CD133：92.4±1.3%。未与抗CD133 mAb结合的聚乙二醇化AuNPs主要聚集在细胞膜处，而与抗CD133 mAb结合的纳米颗粒则包含在细胞核区域内。抗CD133 mAb的结合由于特异性抗原-抗体结合相互作用而促进了特异性细胞内摄取。对HCT116细胞的细胞毒性研究表明，聚乙二醇化AuNPs和聚乙二醇化AuNPs-CD133在任何测试浓度下均未引发任何毒性。同时，相对于未结合抗CD133 mAb缀合物的5-氟尿嘧啶-聚乙二醇化AuNPs处理，5-氟尿嘧啶-聚乙二醇化AuNPs-CD133显著降低了细胞活力（<0.0001）。本研究表明，纳米载体与抗CD133抗体的结合改善了5-氟尿嘧啶对结肠癌细胞的特异性靶向。这些结果表明，将聚乙二醇化AuNPs与抗体和化疗药物同时进行功能化是一种通过靶向药物递送对抗癌症的可行策略。
