Department of Biomedical Sciences, Dental Sciences and Morpho-Functional Imaging, Polyclinic Hospital University, 98125 Messina, Italy.
School of Science and Technology, Centre for Health, Ageing and Understanding of Disease, Nottingham Trent University, Nottingham NG11 8NS, UK.
Int J Mol Sci. 2020 Jul 10;21(14):4891. doi: 10.3390/ijms21144891.
The graphene road in nanomedicine still seems very long and winding because the current knowledge about graphene/cell interactions and the safety issues are not yet sufficiently clarified. Specifically, the impact of graphene exposure on gene expression is a largely unexplored concern. Herein, we investigated the intracellular fate of graphene (G) decorated with cyclodextrins (CD) and loaded with doxorubicin (DOX) and the modulation of genes involved in cancer-associated canonical pathways. Intracellular fate of GCD@DOX, tracked by FLIM, Raman mapping and fluorescence microscopy, evidenced the efficient cellular uptake of GCD@DOX and the presence of DOX in the nucleus, without graphene carrier. The NanoString nCounter™ platform provided evidence for 34 (out of 700) differentially expressed cancer-related genes in HEp-2 cells treated with GCD@DOX (25 µg/mL) compared with untreated cells. Cells treated with GCD alone (25 µg/mL) showed modification for 16 genes. Overall, 14 common genes were differentially expressed in both GCD and GCD@DOX treated cells and 4 of these genes with an opposite trend. The modification of cancer related genes also at sub-cytotoxic G concentration should be taken in consideration for the rational design of safe and effective G-based drug/gene delivery systems. The reliable advantages provided by NanoString technology, such as sensibility and the direct RNA measurements, could be the cornerstone in this field.
在纳米医学中，石墨烯之路似乎仍然漫长而曲折，因为目前关于石墨烯/细胞相互作用和安全问题的知识还不够清楚。具体来说，石墨烯暴露对基因表达的影响是一个尚未充分探索的问题。在此，我们研究了被环糊精（CD）修饰并负载多柔比星（DOX）的石墨烯（G）的细胞内命运，以及参与癌症相关经典途径的基因的调节。通过 FLIM、拉曼映射和荧光显微镜跟踪 GCD@DOX 的细胞内命运，证明了 GCD@DOX 的有效细胞摄取以及 DOX 存在于细胞核中，而没有石墨烯载体。NanoString nCounter™ 平台提供了证据，证明与未经处理的细胞相比，用 GCD@DOX（25 µg/mL）处理的 HEp-2 细胞中有 34 个（700 个中）与癌症相关的差异表达基因（25 µg/mL）。用 GCD 单独处理（25 µg/mL）的细胞显示有 16 个基因发生了变化。总的来说，在 GCD 和 GCD@DOX 处理的细胞中，有 14 个共同的差异表达基因，其中 4 个基因呈相反趋势。在亚细胞毒性 G 浓度下，对与癌症相关的基因的修饰也应该被考虑，以设计安全有效的基于 G 的药物/基因传递系统。NanoString 技术提供的可靠优势，如敏感性和直接的 RNA 测量，可能是该领域的基石。
