Li Shu-jian, Tang Bei-sha, Zhao Guo-hua, Zhang Ru-xu, Xia Kun, Pan Qian
Department of Neurology, Henan Provincial People's Hospital, Zhengzhou, Henan, People's Republic of China.
Zhonghua Yi Xue Yi Chuan Xue Za Zhi. 2011 Oct;28(5):528-31. doi: 10.3760/cma.j.issn.1003-9406.2011.05.012.
To study the effect of Charcot-Marie-Tooth 2L disease causing gene K141N mutation in heat shock protein B8 gene (HSPB8) on cell viability.
By using liposome transfection technique, (wt)HSPB8, (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 degree centigrade 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).
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 (wt)HSPB8 and (K141N)HSPB8 was different from that of control cells. The viability of cells overexpressing (wt)HSPB8 was highest, followed by cells overexpressed with (K141N)HSPB8. The viability of cells tranfected with GFP only was the lowest.
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.
研究夏科-马里-图斯病2L型致病基因热休克蛋白B8基因(HSPB8)K141N突变对细胞活力的影响。
采用脂质体转染技术,将野生型(wt)HSPB8、K141N突变型(K141N)HSPB8真核表达载体及绿色荧光蛋白(GFP)载体分别转染至SHSY-5Y细胞。24小时后,对细胞进行44℃致死性热休克处理40分钟。采用四氮唑蓝比色法(噻唑蓝,MTT)检测各组SHSY-5Y细胞的相对活力。
热休克后,GFP、pEGFP-(wt)HSPB8和pEGFP-(K141N)HSPB8转染组的吸光度值存在显著差异(P<0.05),表明过表达(wt)HSPB8和(K141N)HSPB8的细胞相对活力与对照细胞不同。过表达(wt)HSPB8的细胞活力最高,其次是过表达(K141N)HSPB8的细胞。仅转染GFP的细胞活力最低。
HSPB8在致死性热休克处理下对细胞的保护中可能起重要作用,K141N突变会削弱这种保护作用。