中文摘要：

目的采用已经建立的SHP-2 D61G基因敲入小鼠为研究对象,观察SHP-2激活突变是否对白细胞的黏附、侵袭及细胞因子分泌产生影响,并探讨可能的分子机制。方法常规分离小鼠白细胞,Fibronectin黏附实验检测细胞黏附能力并用结晶紫染色定量,Transwell小室分析细胞迁移,半定量RT-PCR分析白细胞介素-2（IL-2）及肿瘤坏死因子-α（TNF-α）mRNA表达,ELISA法检测细胞培养上清中IL-2及TNF-α浓度,凝胶阻滞实验分析白细胞内NF-κB核内转移情况。结果 SHP-2^D61G/＋激活突变的小鼠白细胞黏附、迁移能力较对照小鼠的白细胞明显增强;突变小鼠白细胞IL-2及TNF-α mRNA表达丰度明显增高,其培养上清中IL-2、TNF-α浓度也相应升高;突变小鼠白细胞NF-κB核内转移明显增多。结论 SHP-2激活突变后可能通过增强的NF-κB信号途径,促进白细胞黏附并侵袭;同时增强其分泌炎症因子的能力,可能介导多器官损伤。

英文摘要：

Objective To investigate whether the leukemia-related activating mutant SHP-2 effect on white blood cells （WBC） motility and its molecular mechanisms.Methods Isolated mouse WBC regularly,cell adhesion and migration induced by fibronectin.IL-2 and TNF-α mRNA expression were analyzed by semi-quantitative RT-PCR.Secreted IL-2 and TNF-α in the supernatant of cultured WBC were quantified using the enzyme-linked immunosorbent assay（ELISA）.NF-κB in WBC nucleus was detected by gel retardation assay（EMSA）.Results Compared with the WBC with wild-type SHP-2,the adhesion or migration ability of WBC with D61G mutant SHP-2 were dramatically increased.The IL-2 and TNF-α mRNA expression level in WBC with mutant SHP-2 were significantly higher than the WT WBC;IL-2 or TNF-α concentration increased in the supernatant of SHP-2 mutant WBC;That NF-κB translocated more into nuclear in SHP-2 D61G mutant WBC induced by IL-3.Conclusion SHP-2 activating mutations may increase WBC NF-κB signaling pathway,and then promote WBC adhesion and infiltration.By increased secreting inflammatory factors,these WBC may injury mouse multiple organs.