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获得慢性粒细胞白血病人原代干祖细胞模型 ,旨在研究慢粒白血病病理发生中信号传导的分子机制。方法 转导b3a2bcr/ablcDNA到正常人CD34+ 细胞中构建人原代慢性粒细胞性白血病模型。采用细胞免疫组化测定了p2 10 BCR/ABL在CD34+ 细胞中的表达 ;细胞粘附、迁移实验进行模型鉴定 ;FACS法测定了p2 10 BCR/ABL转导及对照CD34+ 细胞p2 7kip和MDR 1Pgp的表达。结果 相对于对照的CD34+ 细胞 ,转导了bcr/abl的CD34+ 细胞对纤粘连蛋白 (fibronectin ,FN)的粘附性下降 ;而在FN上的迁移能力增强 ;在低浓度细胞因子或血清条件下表现为凋亡延迟 ;细胞的粒系克隆形成单位数量明显增多 ,这一模型再现了原代慢粒白血病的异常表型特征 ,并以此模型发现p2 10 BCR/ABL转染CD34+ 细胞上调了p2 7Kip水平并可诱导MDR 1Pgp异常表达。结论 该模型适用于慢粒病理发生中的信号分子传导及分子调控研究 ,提示了慢粒耐药的分子机制。
A model of human primary stem and progenitor cells of chronic myelogenous leukemia was obtained to study the molecular mechanisms of signal transduction in the pathogenesis of chronic myeloid leukemia. Methods Human primary chronic myelogenous leukemia was constructed by transduction of b3a2bcr/abl cDNA into normal human CD34+ cells. The expression of p2 10 BCR/ABL in CD34+ cells was detected by cellular immunohistochemistry; cell adhesion and migration assays were performed to identify the model; FACS assays were used to determine p2 10 BCR/ABL transduction and control CD34+ cells p2 7kip and MDR 1 Pgp. expression. Results Compared with control CD34+ cells, CD34+ cells transduced with bcr/abl had a decreased adhesion to fibronectin (FN), but enhanced migration on FN; at low concentrations of cytokine or serum conditions Apoptosis was delayed; the number of clonally formed cells in the cells increased significantly. This model recreates the abnormal phenotypic characteristics of primary chronic myelogenous leukemia, and this model found that p2 10 BCR/ABL transfected CD34+ cells up-regulated p2 7 Kip levels can induce abnormal expression of MDR 1 Pgp. Conclusion This model is suitable for signaling molecular conduction and molecular regulation in the pathogenesis of chronic granules, suggesting the molecular mechanism of chronic granule resistance.