Purpose

This study aimed to endow the cell-penetrating peptide (CPP) S4₁₃-PV with adequate features towards a safe and effective application in cancer gene therapy.

Methods

Peptide/siRNA complexes were prepared with two new derivatives of the CPP S4₁₃-PV, which combine a lauroyl group attached to the N- or C-terminus with a histidine-enrichment in the N-terminus of the S4₁₃-PV peptide, being named C12-H₅-S4₁₃-PV and H₅-S4₁₃-PV-C12, respectively. Physicochemical characterization of siRNA complexes was performed and their cytotoxicity and efficiency to mediate siRNA delivery and gene silencing in cancer cells were assessed in the absence and presence of serum.

Results

Peptide/siRNA complexes prepared with the C12-H₅-S4₁₃-PV derivative showed a nanoscale (ca. 100 nm) particle size, as revealed by TEM, and efficiently mediated gene silencing (37%) in human U87 glioblastoma cells in the presence of 30% serum. In addition, the new C12-H₅-S4₁₃-PV-based siRNA delivery system efficiently downregulated stearoyl-CoA desaturase-1, a key-enzyme of lipid metabolism overexpressed in cancer, which resulted in a significant decrease in the viability of U87 cells. Importantly, these complexes were able to spare healthy human astrocytes.

Conclusions

These encouraging results pave the way for a potential application of the C12-H₅-S4₁₃-PV peptide as a promising tool in cancer gene therapy.

中文翻译：

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富含月桂酰组氨酸的S413-PV肽是癌细胞中高效的基因沉默介体。

目的

这项研究旨在赋予细胞穿透肽（CPP）S4 ₁₃ -PV足够的功能，以便在癌症基因治疗中安全有效地应用。

方法

使用CPP S4 ₁₃ -PV的两个新衍生物制备肽/ siRNA复合物，该衍生物将连接到N-或C末端的月桂酰基与S4 ₁₃ -PV肽N末端的组氨酸富集结合在一起，被C12-H命名₅ -S4 ₁₃ -PV和H ₅ -S4 ₁₃ -PV-C12，分别。进行了siRNA复合物的理化表征，并在无血清和有血清的情况下评估了它们在癌细胞中介导siRNA传递和基因沉默的细胞毒性和效率。

结果

肽/ siRNA复合物制备与C12-H ₅ -S4 ₁₃ -PV衍生物显示出纳米级（约100纳米）的颗粒大小，如通过TEM在人U87成胶质细胞瘤细胞中揭示，且有效地介导的基因沉默（37％）存在30％的血清。此外，新的C12-H ₅ -S4 ₁₃ -PV基于-siRNA递送系统有效地下调硬脂酰CoA去饱和酶-1，脂类代谢的关键酶在癌症中过表达，这导致在U87的活力下降显著细胞。重要的是，这些复合物能够保留健康的人类星形胶质细胞。

结论

这些令人鼓舞的结果铺平了C12-H的潜在应用的方式₅ -S4 ₁₃ -PV肽作为有前途的癌症基因治疗的工具。