构建靶向性载脂蛋白E修饰脂质体介导的HSVtk/丙氧鸟苷系统

Construction of targeting apolipoprotein E-modified liposome-mediated HSVtk/ganciclovir system

背景:自杀基因系统无选择性,不仅能杀伤癌细胞,对正常细胞也有同样作用,所以构建靶向性基因治疗载体成为迫切需要解决的问题。目的:评价载脂蛋白E修饰脂质体(apo E-lipoplexes)介导TK/丙氧鸟苷自杀基因质粒转染对Li-7肝癌细胞的杀伤效果。方法:apo E-lipoplexes包裹p AFP-TK-IRES2-EGFP真核表达质粒转染Li-7细胞,筛选HSVtk稳定表达细胞克隆(Li-7-tk),荧光显微镜观察增强型绿色荧光蛋白表达,Western blotting检测HSVtk基因表达,MTT法评价HSVtk/丙氧鸟苷系统对Li-7肝癌细胞的杀伤作用。结果与结论:自杀基因质粒转染Li-7细胞经筛选得到稳定克隆(Li-7-tk),HSVtk/丙氧鸟苷系统作用于Li-7细胞后,细胞凋亡明显增加(P<0.01)。在甲胎蛋白阳性的Li-7肝癌细胞中,自杀基因载体稳定表达并有效杀灭癌细胞。

BACKGROUND：Suicide gene system without selectivity not only kil s cancer cel s, but also has the same effect on normal cel s, so the construction of targeting vector of gene therapy has become an urgent problem to be solved. OBJECTIVE：To evaluate kil ing effects of apolipoprotein E modified liposomes （apoE-lipoplexes） mediated TK/ganciclovir suicide gene transfection on hepatocel ular carcinoma Li-7 cel s.＆amp;nbsp;METHODS：The apoE-lipoplexes packed pAFP-TK-IRES2-EGFP eukaryotic expression plasmids were transfected into Li-7 cel s. HSVtk stably expressing cel clone （Li-7-tk） was screened. Fluorescence microscopy was used to observe the expression of enhanced green fluorescent protein. HSVtk gene expression was detected using western blotting. MTT method was used to evaluate the kil ing effects of HSVtk/GCV system on Li-7 cel s. RESULTS AND CONCLUSION：Stable clone （Li-7-tk） was obtained through screening suicide gene transfected Li-7 cel s. After HSVtk/ganciclovir system acting on Li-7 cel s, cel apoptosis rate was significantly increased （P〈0.01）. In alpha fetoprotein-positive hepatocel ular carcinoma cel line Li-7, suicide gene vector expresses stably and effectively kil s cancer cel s.