- Author:
Shu-jian LI
1
;
Bei-sha TANG
;
Guo-hua ZHAO
;
Ru-xu ZHANG
;
Kun XIA
;
Qian PAN
Author Information
- Publication Type:Journal Article
- MeSH: Cell Line, Tumor; Cell Survival; genetics; Charcot-Marie-Tooth Disease; genetics; metabolism; Gene Expression Regulation; Genetic Vectors; genetics; Heat-Shock Proteins; genetics; metabolism; Humans; Mutation; genetics; Protein-Serine-Threonine Kinases; genetics; metabolism
- From: Chinese Journal of Medical Genetics 2011;28(5):528-531
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo study the effect of Charcot-Marie-Tooth 2L disease causing gene K141N mutation in heat shock protein B8 gene (HSPB8) on cell viability.
METHODSBy 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).
RESULTSThere 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.
CONCLUSIONHSPB8 may play an important role in the protection of cells under lethal heat shock treatment, and the K141N mutation can impair the protective effect.