Therapeutic targeting of siRNA/anti-cancer drug delivery system for non-melanoma skin cancer. Part I: Development and gene silencing of JAK1siRNA/5-FU loaded liposome nanocomplexes

• Published in: European journal of pharmaceutics and biopharmaceutics 2024-10, Vol.203, p.114432, Article 114432
• Main Authors: Aslan, Minela, Ozturk, Sukru, Shahbazi, Reza, Bozdemir, Özlem, Dilara Zeybek, Naciye, Vargel, İbrahim, Eroğlu, İpek, Ulubayram, Kezban
• Format: Article
• Language: English
• Subjects: 5-FU; Antineoplastic Agents - administration & dosage; Antineoplastic Agents - pharmacology; Cell Line, Tumor; Cell Survival - drug effects; Drug Delivery Systems - methods; Drug Liberation; Fluorouracil - administration & dosage; Fluorouracil - pharmacology; Gene Silencing - drug effects; Humans; JAK1 gene silencing; Janus kinase 1; Janus Kinase 1 - genetics; Janus Kinase 1 - metabolism; Liposomes; Nanoparticles - chemistry; NMSC cancer therapy; Non-melanoma skin cancer; RNA Interference; RNA, Small Interfering - administration & dosage; RNA, Small Interfering - genetics; RNA, Small Interfering - pharmacology; siRNA delivery; Skin Neoplasms - drug therapy; Skin Neoplasms - genetics; Skin Neoplasms - pathology
• ISSN: 0939-6411; 1873-3441; 1873-3441
• DOI: 10.1016/j.ejpb.2024.114432

[Display omitted] •Novel liposome nanocomplexes were prepared using the thin-film hydration method.•Liposome nanocomplexes containing JAK1 siRNA and 5- Fluorouracil exhibited uniform size and stability and maintained high cell viability.•Liposome nanocomplexes effectively internalized within A-431 cells, achieving a 50 % reduction in JAK1 gene expression.•The silencing of JAK1 gene expression led to a 37 % reduction in pERK protein levels. Non-melanoma skin cancer (NMSC) is one of the most prevalent cancers, leading to significant mortality rates due to limited treatment options and a lack of effective therapeutics. Janus kinase (JAK1), a non-receptor tyrosine kinase family member, is involved in various cellular processes, including differentiation, cell proliferation and survival, playing a crucial role in cancer progression. This study aims to provide a more effective treatment for NMSC by concurrently silencing the JAK1 gene and administering 5-Fluorouracil (5-FU) using liposome nanocomplexes as delivery vehicles. Utilizing RNA interference (RNAi) technology, liposome nanocomplexes modified with polyethylene imine (PEI) were conjugated with siRNA molecule targeting JAK1 and loaded with 5-FU. The prepared formulations (NL-PEI) were characterized in terms of their physicochemical properties, morphology, encapsulation efficiency, in vitro drug release, and stability. Cell cytotoxicity, cell uptake and knockdown efficiency were evaluated in human-derived non-melanoma epidermoid carcinoma cells (A-431). High contrast transmission electron microscopy (CTEM) images and dynamic light scattering (DLS) measurements revealed that the nanocomplexes formed spherical morphology with uniform sizes ranging from 80-120 nm. The cationic NL-PEI nanocomplexes successfully internalized within the cytoplasm of A-431, delivering siRNA for specific sequence binding and JAK1 gene silencing. The encapsulation of 5-FU in the nanocomplexes was achieved at 0.2 drug/lipid ratio. Post-treatment with NL-PEI for 24, 48 and 72 h showed cell viability above 80 % at concentrations up to 8.5 × 101 µg/mL. Notably, 5-FU delivery via nanoliposome formulations significantly reduced cell viability at 5-FU concentration of 5 µM and above (p