中文摘要：

目的研究SHP-2酪氨酸磷酸酶激活突变的肥大细胞对白细胞素3（IL3）呈高增殖敏感性的机制。方法从野生型（WT组）、SHP-2杂合突变型（SHP-2D61G/＋）小鼠骨髓中获取髓系细胞,运用细胞集落形成实验观察髓系细胞集落形成能力,用不同剂量的IL30、0.2、2.0、10.0ng/ml）诱导比较两组髓系细胞形成集落数的差异。从髓系细胞中分离培养出肥大细胞,MTS法观察肥大细胞在IL3诱导下的细胞增殖性,Westernblot比较肥大细胞中Erk、Akt的表达及磷酸化水平,免疫共沉淀和Westernblot检测肥大细胞中SHP-2与Gab2结合能力。结果 SHP-2D61G/＋组比WT组的髓系细胞集落形成能力增强;IL3诱导下的肥大细胞在SHP-2D61G/＋组显示了更高的增殖活性;SHP-2D61G/＋组肥大细胞中Erk和Akt的磷酸化水平较WT组明显升高;SHP-2D61G/＋杂合突变后与Gab2结合能力增高。结论 SHP-2D61G/＋的肥大细胞对IL3呈高增殖敏感性,其分子机制可能与SHP-2D61G/＋与Gab2的结合能力增强,从而进一步参与对IL3介导的Ras-Erk、PI3K-Akt信号通路的正调控作用有关。

英文摘要：

Objective To study the mechanism of the mast cell of activating mutant SHP-2 tyrosine phosphatase shows high proliferation sensitivity to IL3. Methods Myeloid cells were obtained from marrow of WT and SHP-2D61G/ ＋ mutant mice. The colony-forming assay（ CFA） was used to observe the colony-forming ability of myeloid cells,and compared the colony-forming ability of two kinds of myeloid cells under different doses of IL3（ 0,0. 2, 2. 0,10. 0 ng/ml） . Mast cells were isolated and cultured from myeloid cells,and MTS assay was used to observe different proliferation of mast cells to IL3 . Western bolt was applied to exam the expression and phosphorylation of Erk and Art in the mast cells,and immunoprecipitation were used to exam the binding capacity of SHP-2 with Gab-2. Results The colony-forming ability of bone marrow cells increased and the proliferative of bone marrow-derived mast cells to IL3 were more obviously in the SHP-2D61G/ ＋ than in the WT. The phosphorylation level of Erk and Akt was higher in the SHP-2D61G/ ＋ than in the WT. The binding capacity of SHP-2D61G/ ＋ with Gab2 improved obviously. Conclusion The bone marrow-derived mast cells shows the high sensitivity to IL3 in the SHP-2D61G/ ＋ . The molecular mechanism may be associated with SHP-2D61G/ ＋ promotes the binding ability of Gab2,and further participates in the IL3-mediated Ras-Erk,PI3K-Akt signaling pathways.