Xue Wenqiang, Du Jinglei, Wei Yingying, Zhou Shizhao, Wang Shicai, Yang Yongzhen, Chen Lin, Yu Shiping
Department of Interventional Therapy, Shanxi Provincial People's Hospital, Taiyuan, 030001, People's Republic of China.
Department of Interventional Therapy, Shanxi Province Cancer Hospital/ Shanxi Hospital Affiliated to Cancer Hospital, Chinese Academy of Medical Sciences/ Cancer Hospital Affiliated to Shanxi Medical University, Taiyuan, 030001, People's Republic of China.
Int J Nanomedicine. 2025 May 28;20:6831-6851. doi: 10.2147/IJN.S521972. eCollection 2025.
Small interfering RNA (siRNA) provides a new method for anti-tumor therapy by targeting Survivin gene in liver cancer. However, free Survivin siRNA is easily degraded and cleared by endonucleases or macrophages in the blood circulation. Therefore, it is urgent to construct a safe and efficient delivery system to achieve effective transfection of Survivin siRNA.
In this study, cationic carbon dots (CCDs) with active targeting ability (FA-PEI-CDs) were designed and prepared using carbon dots (CDs), polyethyleneimine (PEI), and folic acid (FA). Then, Survivin siRNA was loaded by electrostatic adsorption to design a gene-loaded complex with gene silencing effect (FA-PEI-CDs@Survivin siRNA).
As FA-PEI-CDs had good dispersibility, an average diameter of about 21.71 nm and a zeta potential of 5.11 mV. They had good proton buffering capacity and excellent biocompatibility and could not cause erythrocyte hemolysis and thrombosis. When FA-PEI-CDs were mixed with Survivin siRNA at a mass ratio of 2:1, they can completely load siRNA without being interfered by polyanion in vivo and avoid the degradation of siRNA by serum or intracellular nuclease, which significantly increased the circulation time of siRNA in blood. Meantime, when the mass ratio of FA-PEI-CDs to Survivin siRNA was 3:1, the maximum transfection efficiency was 22.8% and 28.5% in HL-7702 cells and HepG2 cells, respectively. In vitro cell experiments confirmed that the gene complex can specifically kill tumor cells without damaging normal liver cells. In vivo tumor inhibition experiments further confirmed that FA-PEI-CDs@Survivin siRNA can cause tumor cell necrosis and reduce the expression of Survivin protein.
In summary, FA-PEI-CDs can carry Survivin siRNA to achieve tumor gene silencing therapy, which will expand the treatment of liver cancer and provide a new idea for carbon nanomaterials in biological genetic engineering.
小干扰RNA（siRNA）通过靶向肝癌中的Survivin基因提供了一种新的抗肿瘤治疗方法。然而，游离的Survivin siRNA在血液循环中容易被核酸内切酶或巨噬细胞降解和清除。因此，迫切需要构建一种安全有效的递送系统，以实现Survivin siRNA的有效转染。
在本研究中，使用碳点（CDs）、聚乙烯亚胺（PEI）和叶酸（FA）设计并制备了具有主动靶向能力的阳离子碳点（CCDs，即FA-PEI-CDs）。然后，通过静电吸附加载Survivin siRNA，设计出具有基因沉默效果的基因负载复合物（FA-PEI-CDs@Survivin siRNA）。
FA-PEI-CDs具有良好的分散性，平均直径约为21.71 nm，zeta电位为5.11 mV。它们具有良好的质子缓冲能力和优异的生物相容性，不会引起红细胞溶血和血栓形成。当FA-PEI-CDs与Survivin siRNA以质量比2:1混合时，它们可以完全负载siRNA，不受体内聚阴离子干扰，避免siRNA被血清或细胞内核酸酶降解，这显著增加了siRNA在血液中的循环时间。同时，当FA-PEI-CDs与Survivin siRNA的质量比为3:1时，在HL-7702细胞和HepG2细胞中的最大转染效率分别为22.8%和28.5%。体外细胞实验证实，该基因复合物可以特异性杀死肿瘤细胞而不损伤正常肝细胞。体内肿瘤抑制实验进一步证实，FA-PEI-CDs@Survivin siRNA可导致肿瘤细胞坏死并降低Survivin蛋白的表达。
综上所述，FA-PEI-CDs可以携带Survivin siRNA实现肿瘤基因沉默治疗，这将拓展肝癌的治疗方法，并为碳纳米材料在生物基因工程中的应用提供新思路。
