Department of Biotechnology and Cell Biology, Medical College, University of Information Technology and Management in Rzeszow, St. Sucharskiego 2, Rzeszow 35-225, Poland.
Department of Biotechnology and Cell Biology, Medical College, University of Information Technology and Management in Rzeszow, St. Sucharskiego 2, Rzeszow 35-225, Poland.
Biomed Pharmacother. 2022 Jun;150:113085. doi: 10.1016/j.biopha.2022.113085. Epub 2022 May 10.
The surface of silver nanoparticles (AgNPs) is characterized by high reactivity resulting in prooxidative and cytotoxic properties. These effects are observed both in normal and in cancer cells, which overexpress the Epidermal Growth Factor Receptor (EGFR). In our previous paper, we have demonstrated that, with the use of liposomes labeled with the Epidermal Growth Factor (EGF), it is possible to direct the toxic effect of AgNPs in EGFR-overexpressing cells. Unfortunately, the mechanism of uptake and toxicity induction by such liposomes is still unknown. Therefore, the aim of this study was to determine the impact of EGF-LipoAgNPs on certain genes related to endocytosis and toxicity induction by such liposomes in human lung (A549) and tongue (SCC-15) cancer cells. The siRNA knock-out gene method was used in this study to determine the engagement of EGFR in this process. The confocal microscopy study revealed that the number of liposomes in the cytoplasm of the A549 and SCC-15 cells was lowered by 51.99 × 10 RFU and 138.50 × 10 RFU, respectively, proving the crucial role of EGFR in the liposome uptake. Moreover, the expression of the SHH and ATM genes was significantly increased, whereas the expression of the NRF2 gene was decreased after the treatment with EGF-LipoAgNPs and native AgNPs. Furthermore, the expression of the CLTC, AP2M1, CAV1, and SH3GLB1 genes indicated that the tested liposomes are uptaken via the clathrin-dependent pathway with engagement of the AP-2 complex and endophilin in this process. Summarizing, the created targeted delivery system of AgNPs causes an increase in the prooxidative and toxic effect of such NPs and has an impact on endocytosis regulatory genes, especially those related to the clathrin-mediated endocytosis.
银纳米粒子（AgNPs）的表面具有高反应性，导致其具有促氧化和细胞毒性。这些效应在正常细胞和癌细胞中都有观察到，而癌细胞过度表达表皮生长因子受体（EGFR）。在我们之前的论文中，我们已经证明，使用标记有表皮生长因子（EGF）的脂质体，可以将 AgNPs 的毒性作用导向 EGFR 过表达的细胞。不幸的是，这种脂质体的摄取和毒性诱导机制仍不清楚。因此，本研究的目的是确定 EGF-LipoAgNPs 对人肺（A549）和舌（SCC-15）癌细胞中与内吞作用和毒性诱导相关的某些基因的影响。在这项研究中，我们使用 siRNA 敲除基因方法来确定 EGFR 在这个过程中的参与。共聚焦显微镜研究表明，A549 和 SCC-15 细胞细胞质中的脂质体数量分别减少了 51.99×10 RFU 和 138.50×10 RFU，证明了 EGFR 在脂质体摄取中的关键作用。此外，在用 EGF-LipoAgNPs 和原生 AgNPs 处理后，SHH 和 ATM 基因的表达显著增加，而 NRF2 基因的表达减少。此外，CLTC、AP2M1、CAV1 和 SH3GLB1 基因的表达表明，测试的脂质体通过网格蛋白依赖性途径被摄取，在此过程中涉及 AP-2 复合物和内吞作用。总之，所创建的 AgNPs 靶向递送系统导致这些 NPs 的促氧化和毒性作用增加，并对参与内吞作用的内吞调节基因产生影响，特别是与网格蛋白介导的内吞作用相关的基因。
