Establishment of cell model for determining activity of insulin receptor kinase by STAT5b responsive reporter gene
- VernacularTitle:STAT5b靶控报告基因检测胰岛素受体激酶活性细胞模型的建立
- Author:
Qingyuan DAI
;
Xianggui CHEN
;
Xiao YANG
;
Jing LUO
- Publication Type:Journal Article
- Keywords:
insulin receptor kinase;
STAT5b;
reporter gene;
cell model
- From:
Chinese Pharmacological Bulletin
1987;0(02):-
- CountryChina
- Language:Chinese
-
Abstract:
Aim To develop a simple and rapid method to monitor insulin receptor kinase activity and provide a novel cell-based model for screening anti-diabetes drugs.Methods CHO cells were co-transfected by plasmids which respectively contained insulin receptor gene,STAT5b gene and luciferase gene driven by STAT5 response elements.The expression of exogenous gene in transfected cells was examined by RT-PCR.The transfected cells were treated by insulin,and then the concentration and time-dependent response of luciferase expression to insulin induction was examined.Moreover,the specificity was identified by AG1024 treatment and PTP1B gene transfection.Results Expressions of insulin receptor and STAT5b were detected in the transfected CHO cells.The expression of luciferase in transfected cells was induced by insulin in concentration and time-dependent way.The maximal induction fold was 6.25.Moreover,the inducible expression of luciferase by insulin could be specifically blocked by tyrphostin AG1024,an inhibitor of insulin receptor kinase,or co-transfected PTP1B gene.Conclusions The insulin receptor kinase activity can be detected by expression of reporter gene with high sensitivity and specificity in this cell model,and with potential value in high throughput screening for insulin receptor activators and sensitizers.
