Zhang Ming-Zhen, Li Cheng, Fang Bi-Yun, Yao Ming-Hao, Ren Qiong-Qiong, Zhang Lin, Zhao Yuan-Di
Britton Chance Center for Biomedical Photonics at Wuhan National Laboratory for Optoelectronics-Hubei Bioinformatics & Molecular Imaging Key Laboratory, Department of Biomedical Engineering, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 430074, People's Republic of China.
Nanotechnology. 2014 Jun 27;25(25):255102. doi: 10.1088/0957-4484/25/25/255102. Epub 2014 Jun 4.
Incorporating ligands with nanoparticle-based carriers for specific delivery of therapeutic nucleic acids (such as antisense oligonucleotides and siRNA) to tumor sites is a promising approach in anti-cancer strategies. However, nanoparticle-based carriers remain insufficient in terms of the selectivity and transfection efficiency. In this paper, we designed a dual receptor-targeted QDs gene carrier QD-(AS-ODN+GE11+c(RGDfK)) which could increase the cellular uptake efficiency and further enhance the transfection efficiency. Here, the targeting ligands used were peptides GE11 and c(RGDfK) which could recognize epidermal growth factor receptors (EGFR) and integrin ανβ3 receptors, respectively. Quantitative flow cytometry and ICP/MS showed that the synergistic effect between EGFR and integrin ανβ3 increased the cellular uptake of QDs carriers. The effects of inhibition agents showed the endocytosis pathway of QD-(AS-ODN+GE11+c(RGDfK)) probe was mainly clathrin-mediated. Western blot confirmed that QD-(AS-ODN+GE11+c(RGDfK)) could further enhance gene silencing efficiency compared to QD-(AS-ODN+GE11) and QD-(AS-ODN+c(RGDfK)), suggesting this dual receptor-targeted gene carrier achieved desired transfection efficiency. In this gene delivery system, QDs could not only be used as a gene vehicle but also as fluorescence probe, allowing for localization and tracking during the delivery process. This transport model is very well referenced for non-viral gene carriers to enhance the targeting ability and transfection efficiency.
将配体与基于纳米颗粒的载体相结合，用于将治疗性核酸（如反义寡核苷酸和小干扰RNA）特异性递送至肿瘤部位，是抗癌策略中一种很有前景的方法。然而，基于纳米颗粒的载体在选择性和转染效率方面仍存在不足。在本文中，我们设计了一种双受体靶向量子点基因载体QD-(AS-ODN+GE11+c(RGDfK))，它可以提高细胞摄取效率，并进一步提高转染效率。这里使用的靶向配体是肽GE11和c(RGDfK)，它们分别可以识别表皮生长因子受体（EGFR）和整合素ανβ3受体。定量流式细胞术和电感耦合等离子体质谱显示，EGFR和整合素ανβ3之间的协同作用增加了量子点载体的细胞摄取。抑制剂的作用表明，QD-(AS-ODN+GE11+c(RGDfK))探针的内吞途径主要是网格蛋白介导的。蛋白质印迹证实，与QD-(AS-ODN+GE11)和QD-(AS-ODN+c(RGDfK))相比，QD-(AS-ODN+GE11+c(RGDfK))可以进一步提高基因沉默效率，表明这种双受体靶向基因载体实现了所需的转染效率。在这个基因递送系统中，量子点不仅可以用作基因载体，还可以用作荧光探针，以便在递送过程中进行定位和追踪。这种转运模型对于非病毒基因载体提高靶向能力和转染效率具有很好的参考价值。
