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目的 探讨阳离子脂质体 L ipofectin TM对 WT1反义寡核苷酸 (反义核酸 )细胞摄入和胞内分布的影响。 方法 采用阳离子脂质体介导 WT1反义核酸转染 K5 6 2细胞 ,应用流式细胞仪、荧光显微镜分别观察反义核酸的细胞摄入和胞内分布 ,RT- PCR检测处理前后 WT1m RNA表达水平。 结果 (1)脂质体提高 K5 6 2细胞对反义核酸的摄入 ,细胞摄入率达 70 .9% ,平均荧光强度提高 6倍以上 ;脂质体介导转染反义核酸时其细胞浆中类核内体或溶酶体中的点状结构较非脂质体处理时大 ,胞核内有明显的荧光物质分布 ;(2 )脂质体介导 WT1反义核酸可下调 WT1m RNA水平。 结论 阳离子脂质体可提高反义核酸的细胞摄入并改变其胞内分布 ,这可能是它增强反义核酸生物活性的机制。
Objective To investigate the effect of cationic liposome L ipofectin (TM) on cellular uptake and intracellular distribution of WT1 antisense oligonucleotide (antisense oligonucleotide). Methods The antisense oligonucleotides of WT1 were transfected into K562 cells using cationic liposomes. The cellular uptake and intracellular distribution of antisense oligonucleotides were detected by flow cytometry and fluorescence microscopy. The expression of WT1mRNA was detected by RT-PCR Level. Results (1) Liposomes increased the uptake of antisense nucleic acid by K562 cells, the cell uptake rate reached 70.9% and the average fluorescence intensity increased more than 6 times. Liposome-mediated transfection of antisense nucleic acids The cytoplasmic endoplasmic reticulum endoplasmic or lysosome in the punctate structure than when non-liposomes treated with significant distribution of fluorescent substance in the nucleus; (2) liposome-mediated WT1 antisense nucleic acid can down-regulate WT1m RNA level. Conclusion Cationic liposomes can increase cellular uptake of antisense nucleic acids and change their intracellular distribution, which may be the mechanism by which cationic liposomes enhance the biological activity of antisense nucleic acids.