高表达衔接蛋白Src羧基端激酶结合蛋白对Jurkat细胞形态及功能的影响

背景：衔接蛋白分子的联动和协同有效调控T细胞的信号转导，形成级联放大或限制，最终实现T细胞复杂的免疫功能。蛋白分子Src羧基端激酶结合蛋白正是衔接蛋白的一种，其主要是通过负反馈调节Src家族激酶活性。而这种负反馈调节作用依赖于Src羧基端激酶结合蛋白的Y317，可能涉及Src羧基端激酶的SH2结构域。目的：探索衔接蛋白Cbp对Jurkat细胞形态及功能的作用。方法：构建仅表达增强绿色荧光蛋白的融合蛋白和Src羧基端激酶结合蛋白-增强绿色荧光蛋白的融合蛋白的病毒颗粒。将Jurkat细胞分为未转染组、阴性对照组和Src羧基端激酶结合蛋白组，后2组转染仅表达增强绿色荧光蛋白的病毒和转染Src羧基端激酶结合蛋白-增强绿色荧光蛋白的病毒。结果与结论：细胞转染效率大于95%， Src羧基端激酶结合蛋白定位于细胞膜上。转染Src羧基端激酶结合蛋白-增强绿色荧光蛋白病毒的Jurkat细胞，皱缩细胞增多，大小均一性较差，细胞坏死凋亡的数量增加，细胞中Src羧基端激酶结合蛋白基因表达上调，Src羧基端激酶表达下降，而酪氨酸蛋白激酶表达增加。提示Jurkat细胞转染Src羧基端激酶结合蛋白后，可使细胞存活率下降，细胞信号转导功能减弱。 中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Effects of high expression of Csk-binding protein on morphology and biological function of Jurkat cells

BACKGROUND:The linkage and synergistic effect of adaptor proteins can effectively regulate signal transduction of T cells, which can form a limit or amplification cascade to realize the complex immune function of T cells. C-terminal Src kinase (Csk)-binding protein (Cbp) is an adaptor protein, which mainly exert the negative feedback regulation of Src kinase activity. This negative feedback effect depends on Y317 of Cbp, which may be involved in the SH2 domain of Csk.OBJECTIVE:To explore the effects of high expression of Cbp on ultrastructure and related biological function of Jurkat cells. METHODS:The virus particles were constructed with expressing enhanced green fluorescent protein (EGFP) only and Cbp-EGFP fusion protein to transfect Jurkat cells. There were untransfected group (Jurkat group), negative control group (transfected with expression of EGFP virus only), and Cbp group (transfected with Cbp-EGFP virus). RESULTS AND CONCLUSION:Confocal microscope showed that cell transfection efficiency was more than 95% and Cbp was located on the cell membrane. Optical microscope showed after transfection with Cbp-EGFP virus, more Jurkat cells shrunk, with poor size uniformity. Apoptosis detection showed that after transfection with Cbp-EGFP virus, the number of apoptotic and necrotic cells was greatly increased. Cbp mRNA expression was increased, Csk expression was decreased obviously and lymphocyte-specific protein tyrosine kinase expression was increased. So, in Jurkat cells, the high expression of Cbp can decrease the uniformity of cells and increase the necrosis cells, thus inhibiting the signal transduction.