C-KIT基因突变对血液祖细胞免疫表型及增殖的影响  

Effect of C-KIT mutations on immunophenotype and proliferation of hematopoietic progenitors

作　　者：杨颖[1] 刘华[2] 郑君芳[2] 郑帅[2] 史小翠[2] 马骞[2] 赵君朋[1] 

机构地区：[1]首都医科大学医学实验与测试中心蛋白质组学研究测试室,北京100069 [2]首都医科大学基础医学院生物化学与分子生物学系,北京100069

出　　处：《基础医学与临床》2013年第10期1247-1251,共5页Basic and Clinical Medicine

基　　金：国家自然科学基金(30900247);北京市教委面上项目(KM201110025002);首都医科大学科研基金(2012ZR03)

摘　　要：目的探讨不同C-KIT突变影响核心结合因子相关性急性髓系白血病(CBF-AML)发生的分子机制。方法用流式细胞术鉴定稳定转染KIT野生型和突变型的EML细胞表面KIT的表达,比较各细胞系的免疫表型;WST-1法检测各细胞系在IL-3或Epo存在下的增殖状况,Western blot法检测Epo受体的表达。结果外源人KIT在各类稳定转染的EML细胞系表面表达,Del417-419insY和D816V C-KIT突变均引起B220+细胞亚群减少以及Sca-1+细胞亚群增多,但后者作用更显著,二者未影响CD34、Gr-1、Mac-1和Ter119阳性细胞百分率;Del417-419insY突变体可以和IL-3、Epo协同促进细胞增殖,而D816V突变体只与IL-3有协同作用,经检测EML-D816V细胞不表达Epo受体。结论不同C-KIT突变可以不同程度改变造血祖细胞免疫表型,协同其他造血因子促进细胞增殖,这提示其与CBF-AML的发生和临床预后的相关性。Objective To investigate the molecular mechanism through which different C-KIT mutations influence the development of CBF-AML. Methods Assessment of exogenous KIT receptor and differentiation antigens on the sur- face of stably transfected EML cells was performed with flow cytometry; Cell proliferation in the presence or absence of IL-3 or Epo was analyzed using WST-1 cell proliferation assay; EpoR expression in different cell lines was identi- fied with Western Blot assay. Results Human KIT receptor was expressed on the surface of transfected EML cells, De1417~lginsY and D816V mutations led to a decrease in percentage of B220 ＋ cells and an increase in Sca-1 ＋ cells, albeit to a more extent for DS16V mutation. Neither of them influenced positive percentage of CD34, Gr-1, Mac-1 and Terll9 in different cells. De1417419insY mutant can cooperate with IL-3 or Epo to promote cell proliferation, while only IL-3 had a synergic effect with Dal6V mutant. Western blot result confirmed the absence of EpoR expres- sion in EML-DS16V cells. Conclusions Distinct C-KIT mutations may change immunophenotype of hematopoietic progenitors and enhance cell proliferation, suggesting their relationship with development and prognosis of CBF-AML.

关 键 词：C—KIT基因突变 核心结合因子相关性急性髓系白血病 造血祖细胞 免疫表型 增殖 

分 类 号：R733.71[医药卫生—肿瘤]