多功能疗法已成为癌症治疗的创新策略。在这篇研究文章中，我们提出了一种纳米结构脂质载体 (NLC)，专为局部治疗皮肤黑色素瘤而设计，可同时递送 5-FU 和 Bcl-2 siRNA。表征的纳米粒子的直径为 259±9 nm，多分散指数为 0.2，表明尺寸分布均匀。 NLC 主要位于表皮，有效地减少了 5-FU 在皮肤层的全身释放。离体皮肤模型揭示了保护性脂质膜的形成，减少了角质层的脱屑过程，这可能与增加渗透的作用有关。体外测定表明，与非癌细胞相比，A375 黑色素瘤细胞对治疗表现出更高的敏感性，反映了其代谢率的预期差异。 A375 细胞对 NLC 的摄取在 4 小时内达到约 90%。使用 ELISA、Western blot 和 qRT-PCR 分析彻底评估了 Bcl-2 敲低的功效，揭示了含有 5-FU 和 Bcl-2 siRNA（低浓度 --100 pM）的 NLC 制剂具有显着的敲低和协同作用。 ）。值得注意的是，Bcl-2 mRNA 的沉默也会影响 Bcl-2 蛋白家族的其他成员，包括 Mcl-1、Bcl-xl、BAX 和 BAK。观察到的这些蛋白质的调节强烈表明细胞凋亡途径的激活，表明成功抑制了黑色素瘤的生长并预防其体外扩散。© 2024。控释协会。

Multifunctional therapies have emerged as innovative strategies in cancer treatment. In this research article, we proposed a nanostructured lipid carrier (NLC) designed for the topical treatment of cutaneous melanoma, which simultaneously delivers 5-FU and Bcl-2 siRNA. The characterized nanoparticles exhibited a diameter of 259 ± 9 nm and a polydispersion index of 0.2, indicating a uniform size distribution. The NLCs were primarily localized in the epidermis, effectively minimizing the systemic release of 5-FU across skin layers. The ex vivo skin model revealed the formation of a protective lipid film, decreasing the desquamation process of the stratum corneum which can be associated to an effect of increasing permeation. In vitro assays demonstrated that A375 melanoma cells exhibited a higher sensitivity to the treatment compared to non-cancerous cells, reflecting the expected difference in their metabolic rates. The uptake of NLC by A375 cells reached approximately 90% within 4 h. The efficacy of Bcl-2 knockdown was thoroughly assessed using ELISA, Western blot, and qRT-PCR analyses, revealing a significant knockdown and synergistic action of the NLC formulation containing 5-FU and Bcl-2 siRNA (at low concentration --100 pM). Notably, the silencing of Bcl-2 mRNA also impacted other members of the Bcl-2 protein family, including Mcl-1, Bcl-xl, BAX, and BAK. The observed modulation of these proteins strongly indicated the activation of the apoptosis pathway, suggesting a successful inhibition of melanoma growth and prevention of its in vitro spread.© 2024. Controlled Release Society.