IDENTIFICATION OF PI3K/Akt PATHWAY RELATED GENES IN VASCULAR SMOOTH MUSCLE CELLS SUBJECTED TO CYCLIC STRAIN
- VernacularTitle:机械张应变诱导的大鼠血管平滑肌细胞磷脂酰肌醇3激酶/蛋白激酶B信号通路相关基因的筛选
- Author:
Qingping YAO
;
Zhiqiang YAN
;
Ling BAI
;
Zonglai JIANG
- Publication Type:Journal Article
- Keywords:
Mechanical strain;
Vascular smooth muscle cell;
Suppression subtractive hybridization;
PI3K/Akt pathway;
Rat
- From:
Acta Anatomica Sinica
2002;0(06):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To screen out the PI3K/Akt pathway related genes in vascular smooth muscle cells (VSMCs) subjected to cyclic strain. Method VSMCs of rat aorta were subjected to cyclic strain (10 %, 1 Hz) by using a FX-4000T system. Phosphorylation of Akt was examined by Western blotting. Suppression subtractive hybridization (SSH) method was employed to analyze the differently expressed cDNA sequence between the strained VSMCs pretreated with and without wortmannin, a specific inhibitor of PI3K. These fragments were ligated with T vectors, screened through the blue-white screening system to establish cDNA library, and was sequenced and analyzed in GenBank with BLASTN search.Result Level of Akt phosphorylation of VSMCs was significantly enhanced by the mechanical strain compared with the control. Ten different expressed sequence tags (EST) were gained after sequencing 30 clones randomly selected from 54 white clones. There may be six genes related with the mechanical strain and cell signal PI3K/Akt, such as High mobility group box 1 (HMGB1), Neural precursor cells expressed (Nedd4a), Cyclin-dependent kinase 5 (CDK5), Histone deacetylase 3 (HDAC3), Ubiquitin-like 1 (Uble1a) and Heterogeneous nuclear ribonucleoprotein H1 (hnRNP). Conclusion SSH is an effective and reliable approach to screen out the genes related with the mechanical strain and PI3K/Akt pathway in VSMCs.
