腓骨肌萎缩症2L热休克蛋白B8过表达对细胞相对活力的影响  

The effect of HSPB8 gene mutation on cell viability in Charcot-Marie-Tooth disease type 2L

作　　者：李书剑[1] 唐北沙[2] 赵国华[2] 张如旭[2] 夏昆[3] 潘乾[3] 

机构地区：[1]河南省人民医院神经内科,郑州450003 [2]中南大学湘雅医院神经内科 [3]中国医学遗传学国家重点实验室

出　　处：《中华医学遗传学杂志》2011年第5期528-531,共4页Chinese Journal of Medical Genetics

基　　金：基金项目：国家自然科学基金（30300200）;国家863计划项目（2004AA227040）

摘　　要：目的探讨轴索型腓骨肌萎缩症2L（axonal Charcot—Marie-Tooth disease type 2L，CMT2L）K141N突变的热休克蛋白B8（heat shock protein B8，HSPB8）对细胞相对活力的影响。方法应用脂质体转染技术，将野生型HSPB8（wtHSPB8）和K141N突变型HSPB8（K141NHSPB8）分别转染SHSY-5Y细胞，使其过量表达上述蛋白，以表达绿色荧光蛋白（green fluorescent protein，GFP）为阴性对照，转染24h后用44℃致死性热休克处理40min，用四甲基偶氮唑蓝（methylthiazolyl tetrazolium，MTT）比色法检测SHSY-5Y细胞的相对活力。结果未处理组pEGFP-wtHSPB8与pEGFP-K141NHSPB8组间吸光度值比较P〉0．05，而热休克处理组pEGFP1-W1HSPB8与pEGFP-k141NHSPB8组间吸光度值比较P〈0．05，表明热休克处理后各组细胞相对活力差异有统计学意义，热休克处理后过表达wtHSPB8的SHSY-5Y细胞活力最高，过表达K141NHSPB8的细胞次之，过表达GFP空载体的细胞活力最低。结论证实了HSPB8在细胞受到致死性热休克刺激时对于保护细胞活力方面起着重要作用，K141N突变型HSPB8对细胞相对活力的保护作用较野生型HSPB8减弱。Objective To study the effect of Charcot-Marie-Tooth 2L disease causing gene K141N mutation in heat shock protein B8 gene （HSPB8）on cell viability. Methods By using liposome transfection technique,Wt HSPBS,K141N HSPB8 eukaryotic expression vector and green fluorescent protein （GFP） vector were transfected into SHSY-5Y cell, respectively. Twenty-four hours later, the cells were treated with 44℃ lethal heat shock for 40 minutes. The relative viability of SHSY-5Y cells in each group was tested by using tetrazole blue colorimetric method （methyl thiazolyl tetrazolium, MTT）. Results There were significant differences among the light absorption value of GFP, pEGFP-wt HSPB8 and pEGFP-K141N HSPB8 transfected groups after heat shock （P 〈0. 05）, indicating that the relative viability of cells overexpressed with wtHSPB8 and K141NHSPB8 was different from that of control cells. The viability of cells overexpressing wtHSPB8 was highest, followed by cells overexpressed with K141NHSPB8. The viability of cells tranfected with GFP only was the lowest. Conclusion HSPB8 may play an important role in the protection of cells under lethal heat shock treatment, and the K141N mutation can impair the protective effect.

关 键 词：腓骨肌萎缩症 热休克蛋白B8 细胞相对活力 四甲基偶氮唑蓝比色法 

分 类 号：R746.4[医药卫生—神经病学与精神病学]