Id-1介导E2-2对PI3K/Akt信号通路及内皮祖细胞增长的影响

Id-1 mediated E2-2 to regulate PI3K/Akt signaling pathway and influenced endothelial progenitor cell increase

目的探讨分化抑制因子1(Id-1)和转录因子E2-2(E2-2)对PI3K/Akt通路的影响情况,并明确其在内皮祖细胞(EPCs)增长中的作用。方法采用免疫共沉淀检测Id-1及E2-2是否存在相互作用。在EPCs细胞模型中,过表达及干扰Id-1及E2-2,采用CCK-8法检测细胞增长情况,RT-PCR和Western blot技术检测PI3K及Akt表达水平。结果免疫共沉淀检测发现,Id-1及E2-2之间存在蛋白-蛋白相互作用。过表达Id-1可显著下调E2-2表达水平,上调PI3K/Akt表达水平,细胞增长能力增强;过表达E2-2后PI3K/Akt表达降低,细胞增长减弱;共转染Id-1及E2-2则发现,二者可协同影响PI3K/Akt表达水平。结论 Id-1和E2-2之间存在相互作用,Id-1对PI3K/Akt起促进作用,而E2-2则发挥抑制效应,二者协同调控PI3K/Akt信号通路,进而影响EPCs的增长。

Objective To explore the associations between Id-1,E2-2 and PI3K/Akt pathway for increase of EPCs. Methods In the EPCs cell model,we used immune coprecipitation test to detect the association between Id-1 and E2-2.We overexpressed or interfered Id-1 or E2-2,and used CCK 8 method to detect cell increase;RT-PCR and Western blotting technique to detect expression of PI3 K,and Akt levels. Results Immune coprecipitation test found that there was protein-protein interaction between Id-1 and E2-2. Compared with the control group,overexpression of Id-1 could decrease the E2-2 expression level,and increase the expression of PI3K/Akt,which lead to inhanced cell proliferation ability. Overexpressing E2-2 reduced the expression of PI3K/Akt and cell proliferation. Co-transfection Id-1 and E2-2 showed that both proteins could influence the expression of PI3K/Akt level synergetically. Conclusion There was protein-protein interaction between Id-1 and E2-2;Id-1 has positive regulatory role for PI3K/Akt while E2-2 has a negative regulatory role. The Id-1 and E2-2 could coordinate to control the PI3K/Akt signal pathway,thus affecting the increase of the EPCs.