阳离子脂质体LipofectinTM对WT1反义核酸在K562细胞的摄入和胞内分布的影响

目的探讨阳离子脂质体LipofectinTM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K562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WT1 mRNA表达水平. 结果 (1)脂质体提高K562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WT1 mRNA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.

Effects of Cationic Liposome LipofectinTM on the Intracellular Uptake and Distribution of WT1 Antisense Oligodeoxynucleotides in K562 Cells

Objective\ To study the effects of cationic liposome on the uptake and intracellular distribution of antisense oligodeoxynucleotide(PSODN).\ Methods\ The uptake and intracellular localization of FAM\|labelled PSODN(F\|PSODN) mediated by cationic liposome Lipofectin was detected by flow cytometry(FCM) and fluorescence microscope.\ The expression of WT1 mRNA was measured by RT\|PCR.\ Results\ (1)Cationic liposomes enhanced K562 cells taking F\|PSODN by 70 9%,and mean fluorescence intensity was increased six times than the controls.\ K562 cells transfection mediated by cationic liposome showed larger punctuate structures in cytosol and more intensive PSODN nuclear accumulation.\ (2)The expression of WT1 \{mRNA\} was decreased in K562 cells transfected with ASODN and cationic liposome after 72 hours.\ \{Conclusion\}\ Cationic liposomes increased PSODN activities by enhancing the uptake of ODN and altering intracellular distribution of the ODN.