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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 has no selectivity. It not only can kill cancer cells but also has the same effect on normal cells. So constructing targeted gene therapy vectors becomes an urgent problem to be solved. OBJECTIVE: To evaluate the cytotoxic effect of apo E-lipoplexes mediated TK / gonadotrophin gene transfer on Li-7 hepatocarcinoma cells. Methods: Li-7 cells were transfected with eukaryotic expression plasmid pFP-TK-IRES2-EGFP by apo E-lipoplexes. The stable expression of HSVtk cell clone (Li-7-tk) was screened and the expression of enhanced green fluorescent protein The expression of HSVtk gene was detected by Western blotting, and the cytotoxicity of HSVtk / gonadoside on Li-7 hepatoma cells was evaluated by MTT assay. RESULTS AND CONCLUSION: The stable clone (Li-7-tk) was obtained after transfection of suicide gene plasmid into Li-7 cells. The apoptosis of Li-7 cells was significantly increased after HSVtk / ). In alpha-fetoprotein positive Li-7 hepatoma cells, suicide gene vectors stably express and effectively kill cancer cells.