脂质体介导的C-myc反义寡核苷酸在视网膜色素上皮细胞中分布和稳定性的观察

Observation on distribution and stability of C-myc antisense oligonucleotides mediated by liposome in retinal pigment epithelium cell

目的在体外培养的人视网膜色素上皮（ＲＰＥ）细胞中，观察脂质体（Ｌｉｐｏｆｅｃｔａｍｉｎｅ）介导的Ｃ－ｍｙｃ反义寡核苷酸转染效果，为进一步研究增生性玻璃体视网膜病变（ＰＶＲ）的防治提供实验依据。方法将人工合成带有ＦＡＭ荧光标记的反义寡核苷酸（ＡＳ－ＯＤＮ）用脂质体包裹转染体外培养的ＲＰＥ细胞，同时用无脂质体介导的裸ＡＳ－ＯＤＮ作对比，在荧光显微镜下动态观察ＡＳ－ＯＤＮ在ＲＰＥ细胞内的分布和存留时间。结果与无脂质体介导的裸ＡＳ－ＯＤＮ相比，脂质体介导的ＡＳ－ＯＤＮ在细胞内、尤其在胞核的分布明显加强；不仅转染速度快，而且在细胞的存留时间延长１倍以上。结论脂质体能明显增强ＡＳ－ＯＤＮ在ＲＰＥ细胞内的转染效率，有望进一步用于防治ＰＶＲ的研究。

ObjectiveTo investigate the transfection effect of antisense oligonucleotides into human retinal pigment epithelium(RPE) cultured mediated by liposome in vitro for a experimental foundation searching for prevention of PVR(proliferative vitreoretinopathy).MethodsThe synthetic phosphorothioate antisense oligodeoxynucleotides(AS-ODN) labeled by FAM fluorescence were complexed with liposome-Lipofectamine,and transfected RPE cultured in vitro,then their location and lasting time within RPE cell were observed under fluorescent microscope for some time,in contrast to the naked AD-ODN without liposome.ResultsIn contrast with naked AS-ODN absent liposome,liposome-mediated AS-ODN distribution in cytoplasm especially in nuclei was distinctly enhanced,and not only velocity of transfection in the present of liposome was more quick but also intracellular retaining time of AS-ODN prolonged at least two-fold.ConclusionsLiposome-Lipofectamine can increase transfective efficiency of AS-ODN in RPE ,hopefully to be used further in prevention of PVR.