Department of Biopharmaceutics, West China School of Pharmacy, Sichuan University, Chengdu, 610041, PR China; Key Laboratory of Drug-Targeting and Drug Delivery System of the Education Ministry, Sichuan Engineering Laboratory for Plant-Sourced Drug and Sichuan Research Center for Drug Precision Industrial Technology, West China School of Pharmacy Sichuan University, Chengdu, 610041, PR China.
Department of Biopharmaceutics, West China School of Pharmacy, Sichuan University, Chengdu, 610041, PR China; Key Laboratory of Drug-Targeting and Drug Delivery System of the Education Ministry, Sichuan Engineering Laboratory for Plant-Sourced Drug and Sichuan Research Center for Drug Precision Industrial Technology, West China School of Pharmacy Sichuan University, Chengdu, 610041, PR China; Stake Key Laboratory of Natural and Biomimetic Drugs, Peking University, PR China.
Biomed Pharmacother. 2020 Aug;128:110236. doi: 10.1016/j.biopha.2020.110236. Epub 2020 May 21.
Integrin αβ was reported as positive regulators of tumorigenesis and highly expressed in cancer stem cells and kinds of cancers, thus, it is an appealing target for cancer treatment. Nanomedicine with targeting delivery ability has developed rapidly and shown its great therapeutic potential in cancer therapy. Proteins are ideal material for nanomedicine regarding to their excellent biocompatibility, and protein-only self-assembled nanoparticles technology provides a robust method to produce protein nanoparticles. Pro-apoptotic proteins or peptides, such as BAK, have attracted increasing attention in the inhibition of tumor growth. However, the self-assembled nanoparticles of BAK targeting to integrin αβ over-expressed tumor cells need to be investigated. In this study, we designed recombinant proteins with BH3 BAK as active domain and RGD peptides as targeting ligands to self-assemble into protein nanoparticles (named as PN2-1 et al.), then experimentally evaluated the nanoparticle size, fluorescence feature, stability, targeting ability and cytotoxicity to tumor cells in vitro. The results showed that the protein nanoparticles containing RGD peptides had a uniform particle size with an diameter of approximately 23 nm. PN2-1 had notable inhibition to cell proliferation of C6 cells, C26 cells and MCF-7 cells, with a lower IC than the nanoparticles which only had BAK motif without RGD peptide. PN2-1 had higher cellular uptake into C6 cells than MCF-7 cells. Our results demonstrate that the RGD peptide could enhance the cytotoxicity of BAK nanoparticles to tumor cells and increase their tumor targeting ability. This study provides an insight into the design and development of integrin αβ targeting protein nanoparticle for cancer treatment.
整合素 αβ 被报道为肿瘤发生的正调控因子，在癌症干细胞和多种癌症中高度表达，因此，它是癌症治疗的一个有吸引力的靶点。具有靶向递药能力的纳米医学发展迅速，在癌症治疗中显示出巨大的治疗潜力。蛋白质由于其优异的生物相容性，是纳米医学的理想材料，而仅由蛋白质自组装的纳米粒技术为制备蛋白质纳米粒提供了一种强大的方法。促凋亡蛋白或肽，如 BAK，在抑制肿瘤生长方面引起了越来越多的关注。然而，针对整合素 αβ 过表达肿瘤细胞的 BAK 靶向自组装纳米粒仍有待研究。在本研究中，我们设计了具有 BH3 BAK 作为活性结构域和 RGD 肽作为靶向配体的重组蛋白，自组装成蛋白质纳米粒（命名为 PN2-1 等），然后在体外实验中评估了纳米粒的粒径、荧光特性、稳定性、靶向能力和对肿瘤细胞的细胞毒性。结果表明，含有 RGD 肽的蛋白质纳米粒具有均匀的粒径，直径约为 23nm。PN2-1 对 C6 细胞、C26 细胞和 MCF-7 细胞的增殖有明显的抑制作用，其 IC 50 值低于仅含有 BAK 基序而不含 RGD 肽的纳米粒。PN2-1 进入 C6 细胞的细胞摄取量高于 MCF-7 细胞。我们的结果表明，RGD 肽可以增强 BAK 纳米粒对肿瘤细胞的细胞毒性，并提高其肿瘤靶向能力。本研究为设计和开发用于癌症治疗的整合素 αβ 靶向蛋白纳米粒提供了思路。
