文献：Efficient Delivery of an Antisense Oligodeoxyribonucleotide Formulated in Folate Receptor-targeted Liposomes

作者：SHIH-JIUAN CHIU, GUIDO MARCUCCI and ROBERT J. LEE

文献链接：https://ar.iiarjournals.org/content/26/2A/1049.short

摘要：

Background: Folate receptors (FRs) are cellular surface markers for numerous solid tumors and myeloid leukemias. The aim of this study was to develop an antisense oligodeoxyribonucleotide (ODN) carrier targeting FR-overexpressing cancer cells using folate (FA) as the targeting moiety. G3139, a phosphorothioate antisense ODN against human bcl2 mRNA, was evaluated in this study. Materials and Methods: G3139-containing liposomes were prepared using an ethanol dilution method. For the targeted formulation, 0.5 mol% of folate-PEG-DSPE was incorporated as a targeting ligand into cationic liposomes composed of DC-Chol/egg PC/PEG-DSPE at 25:65:10 mol/mol. Particle size and surface charge were measured and cellular uptake was assessed by fluorescence microscopy and flow cytometry. The ODN-containing formulations were evaluated in FR+ KB cells for Bcl2 down-regulation measured by Western blot. The cytotoxicity of the formulations was determined by MTT assay. Results: The G3139-containing liposomes had an average diameter of 80-90 nm with high ODN entrapment efficiency (70-80%). Incorporation of the folate ligand did not significantly alter the particle size and entrapment efficiency. The formulation exhibited colloidal stability in a serum-containing environment. In uptake studies, the folate-targeted formulation showed ligand concentration-dependent uptake that was up to 6-fold more efficient than that of the non-targeted formulation (p<0.05). The uptake could be blocked by an excess amount of free folate, thus indicating an FR-dependent mechanism. Conclusion: FR-targeted G3139-containing liposomes showed promising transfection activity in KB cells. FR-targeted formulations were capable of specific targeting to FR-overexpressing cell lines and optimizing the amount of folate ligand in the liposomal formulation can result in more efficient antisense delivery.

本研究的目的是开发一种反义寡脱氧核糖核苷酸（ODN）载体，以叶酸（FA）为靶向部分，靶向过表达FR的细胞。本研究评估了针对人bcl2 mRNA的硫代反义寡核苷酸G3139。

材料和方法：采用乙醇稀释法制备含G3139的脂质体。对于靶向制剂，以25:65:10 mol/mol的比例将0.5 mol%的叶酸-PEG-DSPE作为靶向配体掺入由DC-Chol/egg PC/PEG-DSPE组成的阳离子脂质体中。测量粒径和表面电荷，并通过荧光显微镜和流式细胞术评估细胞摄取。通过蛋白质印迹法在FR+KB细胞中评估含ODN的制剂的Bcl2下调情况。通过MTT法测定制剂的细胞毒性。

结果：含G3139的脂质体平均直径为80-90nm，ODN包封率高（70-80%）。叶酸配体的掺入没有显著改变粒径和包封效率。该制剂在含血清的环境中表现出胶体稳定性。

在摄取研究中，叶酸靶向制剂显示出配体浓度依赖性摄取，其效率比非靶向制剂高出6倍（p<0.05）。过量的游离叶酸可以阻断摄取，从而表明FR依赖机制。

结论：含FR靶向G3139的脂质体在KB细胞中显示出良好的转染活性。FR靶向制剂能够特异性靶向FR过表达的细胞系，优化脂质体制剂中叶酸配体的量可以导致更有效的反义递送。

相关推荐：

Cy7-DSPE

Cy7-PEG-DOPE

Cy7-PEG-DPPE

DSPE-PEG-Cy7

DSPE-PEG-cRGD

DPPE-PEG-cRGD

DMPE-PEG-cRGD

DOPE-PEG-cRGD

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