Department of Biochemistry and Molecular Biology, University of Rajshahi, Rajshahi, Bangladesh.
Cancer Molecular Pathology of School of Medicine and Dentistry, Menzies Health Institute Queensland, Griffith University, Gold Coast, QLD, Australia.
Methods Mol Biol. 2022;2534:121-133. doi: 10.1007/978-1-0716-2505-7_9.
The discovery of RNA interference (RNAi) has opened a new strategy in cancer therapy, especially by silencing target genes. Pharmacologically it can be achieved by introducing of small (19-21 base pairs) dsRNA molecules known as small interfering RNA (siRNA) targeting interested genes. siRNA mediated gene has been widely investigated for its utility in treating various diseases including cancer. However, the systemic delivery of interested siRNA via non-viral methods remains a major challenge with large numbers of polymeric and liposomal systems being tested. The most effective methods involving cationic liposomes delivery to cells. Nonetheless, systemic delivery of siRNA via cationic lipid particles is often poor due to rapid uptake by reticuloendothelial organs, resulting in decreased delivery of these particles to the site of interest. Polyethylene glycol (PEG) has been used in siRNA-liposomes formulation to minimize reticuloendothelial uptake. Also, PEGylation permits the accumulation of the liposomes-loaded siRNA at the tumor sites with defective vasculatures such as enhanced permeability and retention phenomena. Thus, a simple method to prepare stable PEGylated siRNA-loaded lipid particles could provide better systemic delivery system in treating various cancers, including papillary thyroid carcinoma. Here we illustrate a simple protocol for the formulation of siRNA-loaded lipid particles by hydration of freeze-dried matrix (HFDM) method for effective delivery of target specific siRNA to papillary thyroid carcinoma cells.
RNA 干扰 (RNAi) 的发现为癌症治疗开辟了新的策略，特别是通过沉默靶基因。在药理学上，可以通过引入小 (19-21 个碱基对) dsRNA 分子，即靶向感兴趣基因的小干扰 RNA (siRNA) 来实现。siRNA 介导的基因因其在治疗各种疾病（包括癌症）方面的应用而得到广泛研究。然而，通过非病毒方法系统地递送至感兴趣的 siRNA 仍然是一个主要挑战，大量的聚合物和脂质体系统正在被测试。最有效的方法涉及阳离子脂质体向细胞的递送。尽管如此，由于网状内皮器官的快速摄取，阳离子脂质体介导的 siRNA 的全身递送通常较差，导致这些颗粒向靶部位的递送减少。聚乙二醇 (PEG) 已被用于 siRNA-脂质体配方中，以最小化网状内皮细胞的摄取。此外，PEG 化允许载有 siRNA 的脂质体在诸如通透性增强和保留现象等血管缺陷的肿瘤部位积聚。因此，一种制备稳定的 PEG 化 siRNA 负载脂质体的简单方法可以为治疗各种癌症（包括甲状腺乳头状癌）提供更好的全身递送系统。在这里，我们展示了一种通过冷冻干燥基质水合法 (HFDM) 制备负载 siRNA 的脂质体的简单方案，以有效地将靶向特异性 siRNA 递送至甲状腺乳头状癌细胞。
