SHP-2酪氨酸磷酸酶在DNA损伤性放化疗引发的小鼠骨髓毒中的作用  

Roles and mechanism of SHP-2 tyrosine phosphatase in myeloid cytotoxicity induced by DNA damaging chemotherapy and radiotherapy

作　　者：张梅[1] 李菲菲[1] 郑红[1] 倪芳[1] 瞿成奎[1] 汪思应[1] 

机构地区：[1]安徽医科大学病理生理教研室,合肥230032

出　　处：《安徽医科大学学报》2008年第5期480-484,共5页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:30470735);安徽省人才开发基金(编号:2002Z035);安徽省2006研究实验基地科研带头人培养专项基金;安徽省自然科学基金(编号:070413087);安徽省教育厅自然科学基金项目(编号:2004kj209;KJ2007B035)

摘　　要：目的研究酪氨酸磷酸酶SHP-2在放、化疗引发的骨髓毒的调节作用,初步探讨其可能的分子机制。方法分别从野生型(Wild-type,WT)、SHP-2杂合突变型(SHP-2+/-)及SHP-2纯合突变型(SHP-2-/-)胚胎鼠(9.0~9.5d)的卵黄囊中获取造血祖细胞,用细胞因子IL-3(Interleukin-3,0.5ng/ml)、SCF(stem cell factor,0.5ng/ml)将造血祖细胞定向诱导分化为粒-巨噬造血细胞系(granulocyte-macrophage,GM)。60Coγ射线及化疗药物顺铂(Cisplatin,CDDP)处理后,台盼蓝染色法观察比较三种GM细胞增殖抑制率的差异;集落形成实验(Colony-forming Units,CFU)分析比较卵黄囊造血祖细胞集落形成能力的差异;血细胞计数法检测比较WT小鼠和SHP-2+/-小鼠在照射或化疗后外周血中白细胞、红细胞及血红蛋白水平的变化;Western Blot检测Erk在WT及SHP-2-/-造血细胞中的表达与磷酸化水平。结果在CDDP或60Coγ射线导致的细胞损伤反应中,SHP-2-/-造血细胞的生存能力较SHP-2+/-和WT造血细胞增强;相比WT小鼠,SHP-2+/-组小鼠的白细胞、血红蛋白及红细胞数量降低程度减轻;SHP-2-/-造血祖细胞克隆集落形成能力较SHP-2+/-组及WT组增强;提示SHP-2-/-突变可以减轻CDDP或60Coγ导致的骨髓毒性。Western Blot显示在CDDP损伤反应中,SHP-2-/-抑制Erk的磷酸化。结论SHP-2参与调控放、化疗导致的骨髓毒性,其分子机制可能与SHP-2对Erk活化调节相关。Objective To demonstrate potential roles and mechanism for SHP-2 tyrosine phosphatase in myeloidcytotoxicity induced by DNA damaging chemotherapy and radiotherapy.Methods Mouse embryonic yolk sac hematopoietic progenitor cells were derived from day 9.0~9.5 embryos of SHP-2 homozygous mutant mouse（SHP-2-/-）,SHP-2 heterozygosis mutant mouse（SHP-2＋/-）and wild type（WT）mouse.Then they were committed to granulocyte-macrophage（GM）differentiation after incubated with IL-3（0.5 ng/ml）and SCF（0.5 ng/ml）.These three kinds of GM cell were then treated with cisplatin（CDDP）or ^（60）Coγ-irradiation and subjected to trypan blue staining and colony forming units analysis（CFU）.Haemacytometry was applied to detect the levels of leucocyte,akaryocyte and haematoglobin in WT and SHP-2＋/-mice.Western bolt was applied to exam the expression and phosphorylation of Erk in WT and SHP-2-/-mutation hematopoietic cell.Results Trypan blue staining showed that following DNA damage induced by cisplatin or ^（60）Co γ-irradiation,cell reproductive activity of SHP-2-/-hematopoietic cells were much better than SHP＋/-and WT hematopoietic cells.Levels of leucocyte,akaryocyte and haematoglobin in blood of SHP-2＋/-mutant mice were also higher than WT mice.Colony Forming Units Analysis showed an increased numbers of colony units of SHP-2-/-hematopoietic cell compared with SHP＋/-cell and WT cell,Erk kinase activation was decreased in the SHP-2-/-mutant hematopoietic cells after CDDP treated.Conclusion SHP-2 is involved in regulation of myeloid cytotoxicity induced by DNA damaging chemotherapy and radiotherapy through regulating the Erk-MAP kinase pathways.

关 键 词：蛋白质酪氨酸激酶 骨髓/药物作用 抗肿瘤联合化疗方案/副作用 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学] R331.122[医药卫生—基础医学]