Institute of Life Sciences, Jilin University, Changchun, Jilin 130023, China.
Biomed Res Int. 2013;2013:710502. doi: 10.1155/2013/710502. Epub 2013 Jun 1.
A novel antisense oligonucleotide (ASO) carrier, polyethylenimine conjugated to linoleic acid (PEI-LA), was synthesized and evaluated for delivery of LOR-2501 to tumor cells. LOR-2501 is an ASO targeting ribonucleotide reductase R1 subunit (RRM1). In this study, PEI-LA was synthesized by reacting PEI (Mw ~ 800) with linoleoyl chloride. Gel retardation assay showed complete complexation between PEI-LA and LOR-2501 at N/P ratio above 8. No significant cytotoxicity was observed with these complexes at the tested dosage levels. Interestingly, at N/P ratio of >6, levels of cellular uptake of PEI-LA/LOR-2501 were double that of PEI/LOR-2501 complexes of the same N/P ratio. PEI-LA/LOR-2501 induced downregulation of 64% and 70% of RRM1 at mRNA and protein levels, respectively. The highest transfection activity was shown by PEI-LA/LOR-2501 complexes at N/P ratio of 10. Finally, using pathway specific inhibitors, clathrin-mediated endocytosis was shown to be the principle mechanism of cellular internalization of these complexes. In conclusion, PEI-LA is a promising agent for the delivery of ASOs and warrants further investigation.
一种新型的反义寡核苷酸（ASO）载体，聚亚乙基亚胺与亚油酸（PEI-LA）偶联物，被合成并评估用于将 LOR-2501 递送至肿瘤细胞。LOR-2501 是一种针对核苷酸还原酶 R1 亚单位（RRM1）的 ASO。在这项研究中，PEI-LA 通过使 PEI（Mw~800）与亚油酰氯反应而合成。凝胶阻滞实验表明，在 N/P 比高于 8 时，PEI-LA 与 LOR-2501 之间完全发生了络合。在测试的剂量水平下，这些复合物没有表现出明显的细胞毒性。有趣的是，在 N/P 比大于 6 时，PEI-LA/LOR-2501 的细胞摄取水平是相同 N/P 比的 PEI/LOR-2501 复合物的两倍。PEI-LA/LOR-2501 在 mRNA 和蛋白质水平上分别诱导 RRM1 的下调 64%和 70%。在 N/P 比为 10 时，PEI-LA/LOR-2501 复合物表现出最高的转染活性。最后，使用途径特异性抑制剂表明，网格蛋白介导的内吞作用是这些复合物细胞内化的主要机制。总之，PEI-LA 是一种有前途的 ASO 递送剂，值得进一步研究。
