Department of Molecular Cell Biology, Institute for Cancer Research, Oslo University Hospital, The Norwegian Radium Hospital, Oslo, Norway.
Department of Biosciences, University of Oslo, Oslo, Norway.
Nanotoxicology. 2021 Sep;15(7):865-884. doi: 10.1080/17435390.2021.1924888. Epub 2021 May 28.
Nanoparticles composed of poly(alkyl cyanoacrylate) (PACA) have shown great promise due to their biodegradability and high drug loading capacity. Development of optimal PACA nanocarriers requires detailed analysis of the overall cellular impact exerted by PACA variants. We here perform a comprehensive comparison of cabazitaxel (CBZ)-loaded nanocarriers composed of three different PACA monomers, i.e. poly(n-butyl cyanoacrylate) (PBCA), poly(2-ethylbutyl cyanoacrylate) (PEBCA) and poly(octyl cyanoacrylate) (POCA). The cytotoxicity of drug-loaded and empty PACA nanoparticles were compared to that of free CBZ across a panel of nine cancer cell lines by assessing cellular metabolism, proliferation and protein synthesis. The analyses revealed that the cytotoxicity of all CBZ-loaded PACAs was similar to that of free CBZ for all cell lines tested, whereas the empty PACAs exerted much lower toxicity. To increase our understanding of the toxic effects of these treatments comprehensive MS-based proteomics were performed with HCT116, MDA-MB-231 and PC3 cells incubated with PACA-CBZ variants or free CBZ. Interestingly, PACA-CBZ specifically led to decreased levels of proteins involved in focal adhesion and stress fibers in all cell lines. Since we recently demonstrated that encapsulation of CBZ within PEBCA nanoparticles significantly improved the therapeutic effect of CBZ on a patient derived xenograft model in mice, we investigated the effects of this PACA variant more closely by immunoblotting. Interestingly, we detected several changes in the protein expression and degree of phosphorylation of SRC-pathway proteins that can be relevant for the therapeutic effects of these substances.
基于聚（烷基氰基丙烯酸酯）（PACA）的纳米粒子由于其生物降解性和高载药能力而显示出巨大的应用前景。开发最佳的 PACA 纳米载体需要详细分析 PACA 变体对整体细胞的影响。我们在这里对三种不同 PACA 单体（即聚正丁基氰基丙烯酸酯（PBCA）、聚 2-乙基丁基氰基丙烯酸酯（PEBCA）和聚辛基氰基丙烯酸酯（POCA））组成的载紫杉醇（CBZ）纳米载体进行了全面比较。通过评估细胞代谢、增殖和蛋白质合成，比较了载药和空 PACA 纳米颗粒与游离 CBZ 在 9 种癌细胞系中的细胞毒性。分析表明，所有载 CBZ 的 PACA 的细胞毒性与游离 CBZ 相似，而空 PACA 的毒性要低得多。为了更全面地了解这些处理的毒性作用，我们用 HCT116、MDA-MB-231 和 PC3 细胞进行了基于 MS 的蛋白质组学分析，这些细胞分别与 PACA-CBZ 变体或游离 CBZ 孵育。有趣的是，PACA-CBZ 特异性地导致所有细胞系中与焦点黏附和应激纤维相关的蛋白质水平降低。由于我们最近证明，将 CBZ 封装在 PEBCA 纳米粒子内可显著提高 CBZ 在小鼠来源的异种移植模型中的治疗效果，因此我们通过免疫印迹更仔细地研究了这种 PACA 变体的影响。有趣的是，我们检测到 SRC 通路蛋白的表达和磷酸化程度发生了几种变化，这可能与这些物质的治疗效果有关。
