Nuffield Department of Women's and Reproductive Health, Oxford University John Radcliffe Hospital, Headington, Oxford OX3 9DU, UK.
Department of Engineering Science, Oxford University, Parks Road, Oxford OX1 3PJ, UK.
Biomolecules. 2022 Aug 17;12(8):1129. doi: 10.3390/biom12081129.
Metastatic alveolar rhabdomyosarcoma (aRMS) is an aggressive paediatric cancer with a poor prognosis. Downregulation of critical tumour genes using targeted siRNA remains an obstacle, but association with nanoparticles could help to deliver, protect, target, and enhance penetration. siRNA towards two genes was investigated: (i) Human αB-crystallin (CRYAB) and Heat Shock Protein Family B (Small) Member 2 (HSPB2), and (ii) Keratin 17 (KRT17). A mesoporous silica based nanosystem was linked to siRNA via disulfide bonds and loaded with IR820 dye. Transfection efficiency and signalling was evaluated, and the metabolic effects and cell proliferation were monitored in 2D culture and 3D spheroid models. The bound siRNA was protected from degradation with RNase I for at least 24 h. The delivered siRNA showed significant suppression of viability; 53.21 ± 23.40% for CRYAB and HSPB2 siRNA, and 88.06 ± 17.28% for KRT17 siRNA. After 72 h this increased to >50% cell apoptosis and necrosis. Intracellular total glutathione (GSH) levels were also compared with fibroblasts, and the RMS cell lines showed a several-fold increase. IR820 cellular uptake rate and penetration depth was significantly improved by nanoparticle delivery. Targetted siRNA delivery may pave the way for less invasive and more effective treatments of aRMS.
转移性肺泡横纹肌肉瘤（aRMS）是一种具有不良预后的侵袭性儿科癌症。使用靶向 siRNA 下调关键肿瘤基因仍然是一个障碍，但与纳米颗粒结合可以帮助递药、保护、靶向和增强穿透性。研究了两种基因的 siRNA：（i）人αB-晶体蛋白（CRYAB）和热休克蛋白家族 B（小）成员 2（HSPB2），和（ii）角蛋白 17（KRT17）。介孔二氧化硅纳米系统通过二硫键与 siRNA 连接，并负载 IR820 染料。评估了转染效率和信号转导，并在 2D 培养和 3D 球体模型中监测了代谢效应和细胞增殖。结合的 siRNA 用核糖核酸酶 I 保护至少 24 小时免受降解。递送到细胞内的 siRNA 显著抑制了细胞活力；CRYAB 和 HSPB2 siRNA 的抑制率为 53.21±23.40%，KRT17 siRNA 的抑制率为 88.06±17.28%。72 小时后，细胞凋亡和坏死率增加到超过 50%。还比较了细胞内总谷胱甘肽（GSH）水平与成纤维细胞的水平，RMS 细胞系的 GSH 水平增加了数倍。通过纳米颗粒递送，IR820 的细胞摄取率和穿透深度显著提高。靶向 siRNA 递药可能为侵袭性更小、更有效的 aRMS 治疗铺平道路。
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