【摘要】 目的　观察表皮生长因子受体反义寡核苷酸经转染后 ,在胶质细胞中的通透性及稳定性表达。　方法　将异硫氰酸荧光素标记的硫代磷酸化修饰型及未修饰型的表皮生长因子受体反义寡核苷酸经阳离子脂质体介导或直接转染胶质细胞 ,分别于 15、30 min、1h后在荧光显微镜下观察胶质细胞中荧光分布情况。　结果　直接转染 ,修饰型及未修饰型表皮生长因子受体反义寡核苷酸 30 min时少数胶质细胞胞浆可见点状荧光分布 ,4 h后 ,近 5 0 %的细胞可见荧光分布 ,修饰型可稳定存在于细胞中 3～ 4 h,8h后荧光消失 ;脂质体介导转染 ,表皮生长因子受体反义寡核苷酸 15 min进入胶质细胞 ,4 h后 70 %～ 80 %的胶质细胞可见荧光分布 ,非修饰型 4 h荧光消失 ,修饰型可稳定存在于细胞中 10～ 12 h,14 h后荧光消失。　结论　脂质体包裹的修饰型表皮生长因子受体反义寡核苷酸能进入胶质细胞内并稳定表达更多还原

【Abstract】 Objective To observe the permeability and stability of the transfection of antisense oligonucleotide (ASODN) hybridized epidermal growth factor receptor (EGFR) to retinal glial cells (RG). Methods Phosphorothioate and unmodified EGFR ASODN conjugated with 5′ isothiocyanate (5′ FITC) were encapsulated with or without lipofectin, and then added into human retinal glial cells culture media. The cellular permeability and stability of the transfection were observed by fluorescence microscopy in fixed cells. Results In the absence of lipofectin, phosphorothioate and unmodified EGFR ASODN were found in a few RG cells at 30 minutes, and in about 50% RG cells at 4 hours. Phosphorothioate EGFR ASODN were kept in RG cells for 3 4 hours and disappeared at about 8 hours. In the presence of lipofectin, phosphorothioate and unmodified EGFR ASODN were found in a few RG cells at 15 minutes and about 70% 80% RG cells at 4 hours. Phosphorothioate EGFR ASODN were kept in cells for 10 12 hours, and phosphorothioate and unmodified EGFR ASODN were disappeared at about 14 hours and 4 hours respectively. Conclusion 5′ FITC EGFR ASODN encapsulated with lipofectin could enter RG cells and express stably in RG cells.更多还原