SHP-2酪氨酸磷酸酶激活突变导致小鼠髓系异常增殖  

An activated mutant of SHP-2 tyrosine phosphatase causes murine myeloid abnormal proliferation

作　　者：张薇[1,2] 杨金莲[1] 胡中倩[1] 卢艳敏[1] 储著朗[1] 余科科[1] 瞿成奎[1,3] 汪思应[1] 

机构地区：[1]安徽医科大学病理生理学教研室,安徽合肥230032 [2]安徽医学高等专科学校病理生理学教研室,安徽合肥230601 [3]美国凯斯西储大学,克里夫兰44106

出　　处：《中国病理生理杂志》2011年第4期682-687,共6页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.30873046);安徽省优秀留学人才基金资助项目(No.2009-2010)

摘　　要：目的:观察激活突变SHP-2酪氨酸磷酸酶是否参与髓系异常增殖的发生。方法:以野生型(WT)和SHP-2^(D61G/+)突变型C57BL/6小鼠为研究对象,计数外周血白细胞,比较脾大小,流式细胞术检测外周血及骨髓髓系来源细胞表面标志分子(Mac-1、Gr-1),并统计外周血Mac-1和Gr-1阳性细胞率及骨髓细胞中红系(Ter119)、髓系(Mac-1、Gr-1)、T(CD3)、B(B220)淋巴细胞系的阳性细胞率,观察骨髓造血干/祖细胞的集落形成(CFU)能力,Western blotting检测外周血白细胞经白细胞介素3(IL-3)和5μg/L,刺激后磷酸化的丝氨酸/苏氨酸蛋白激酶B(p-Akt)和磷酸化的细胞外信号调节激酶(p-ERK)表达水平。结果:SHP-2^(D61G/+)突变16周龄组小鼠较WT组外周血白细胞数增多(P<0.05),脾明显增大,同时外周血白细胞的Mac-1和Gr-1阳性细胞率增加(P<0.05),骨髓细胞中Mac-1和Gr-1的阳性细胞率也增多,但红系、淋巴细胞系的变化不明显。同时骨髓中粒-单核细胞集落形成单位(CFU-GM)较正常对照组明显增加,白细胞经IL-3刺激后Akt和ERK蛋白磷酸化水平升高。结论:SHP-2D61G突变可能通过MAPK及PI3K的活化而导致小鼠髓系异常增殖。AIM：To investigate whether an activated mutant of SHP-2 tyrosine phosphatase is involved in abnormal proliferation of murine myeloid.METHODS：Wild-type（WT）and SHP-2^（D61G/＋）mutant mice aged 8 weeks and 16 weeks were used.The number of peripheral blood leukocytes and the spleen sizes were measured by oell counting and weighing methods,respectively.The surface markers（Mac-1 and Gr-1 for myeloid,Ter119 for erythroid,CD3 for T-lymphocyte and B220 for B-lymphocyte）of hematopoitic cells in peripheral blood and bone marrow were detected by flow cytometry.The rate of Mac-1 or Gr-1 positive cells in the peripheral blood and the rate of Mac-1,Gr-1,Terl 19, CD3 or B220 positive cells in bone marrow were analyzed.The ability of colony formation unit（CFU）of the bone marrow was also observed by CFU assay.Finally,the expression of p-Akt and p-ERK in the peripheral blood leukocytes induced by interleukin-3（IL-3,5μg/L）was detected by Western blotting.RESULTS：The number of leukocytes in peripheral blood of 16-week-old mice was more（P〈0.01）and the spleens were bigger in mutant SHP-2^（D61G/＋）mice than those in WT mice.The rate of Mac-1 and Gr-1 positive cells in peripheral blood leukocytes of 16-week-old SHP -2^（D61G/＋）mice were dramatically increased（P〈0.05）.Mac-1 and Gr-1 positive cell rates in bone marrow of SHP- 2^（D61G/＋）mice were much higher（P〈0.05）than those in WT mice and no statistic;significance was found in the erythroid or lymphocyte cells.The number of CFU-GM（represents myeloid）was increased in mutant mice.The expression of p-Akt and p-ERK in peripheral blood leukocytes of mutant mice was significantly enhanced after stimulated with IL-3. CONCLUSION：These results suggest that activated mutant SHP-2 results in the disorder of mouse myeloid proliferation via MAPK and PI3K activation.

关 键 词：SHP-2酪氨酸磷酸酶 激活突变 小鼠 髓系增殖 

分 类 号：R363[医药卫生—病理学]