转染早老素1突变基因SH-SY5Y细胞凋亡的研究Study on the Role of Mutant PS-1 Gene in Alzheimer Disease
冯颖,许贤豪,彭丹涛,张华,乔立艳,张星虎,刘广志,殷剑,国红,王红,李宝林,莫培生
摘要(Abstract)：
目的 探索转染早老素 -突变基因导致 SH-SY5 Y细胞凋亡的机制。方法 用 Xba I和 Acc6 5 I内切酶将携带野生型和突变型 PS-1的 p Bluescript质粒双酶切 ,电泳后纯化其 c DNA片段 ,然后将其克隆到 p CDNA3载体中。由脂质体介导将 p CDNA3空载体、携带有野生型和突变型 PS-1基因的 p CDNA3质粒分别转染到神经母细胞瘤细胞系 SH-SY5 Y细胞中。用蛋白印迹 ( Western blot)杂交检测其表达。用流式细胞仪 ( Coulter EpicsXL)检测在不同情况下不同细胞凋亡的百分率 ,荧光显微镜下观察碘化吡啶 ( PI)、Hoechst3334 2 ( Ho)双染的凋亡细胞的形态 ,并用分子生物学方法检测凋亡细胞的 DNA状况。结果 在无血清培养 2 4 h条件下含有空载体、野生型和突变型 ( A2 4 6 E) PS-1基因的 SY5 Y细胞凋亡百分率依次为 ( 6 .0 3± 2 .0 3)、( 8.34± 2 .4 1 )、( 2 6 .2±4 .6 )。用 SY5 Y细胞作为对照 ,其凋亡百分率为 ( 4 .2 2± 1 .89)。在 Aβ2 5 -35 ( 2 5 μmol/L)诱导下其凋亡百分率依次为 ( 2 4 .4± 3.6 )、( 2 7.7± 2 .5 )、( 5 0 .3± 8.3)。SY5 Y对照组为 ( 1 8.4± 3.1 )。比较显示含突变早老素 1基因细胞的凋亡程度明显升高 ( P<0 .0 0 1 )。结论 早老素 1基因突变导致神经细胞更容易在各种刺激下发生凋亡 ,
关键词(KeyWords)： Alzheimer病;早老素;突变;凋亡
基金项目(Foundation): “九五”国家医学攻关课题基金资助项目 ( 96 -90 6 -0 5 -0 7)
作者(Author): 冯颖,许贤豪,彭丹涛,张华,乔立艳,张星虎,刘广志,殷剑,国红,王红,李宝林,莫培生
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