【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。更多还原

【Abstract】 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.更多还原