CML患者循环内皮细胞中BCR/ABL融合基因表达水平及其临床意义的研究  

Expression of BCR /ABL Fusion Gene in Circulating Endothelial Cells from Chronic Myelogenous Leukemia Patients and Its Clinical Significance

作　　者：吴静怡 黄亮[2] 周剑峰[2] 裴仁治 马俊霞 张丕胜 刘旭辉 杜小红 陈冬 沙科娅 李双月 曹俊杰 

机构地区：[1]浙江省宁波市鄞州人民医院血液内科,浙江宁波315000 [2]华中科技大学同济医学院附属同济医院血液内科,湖北武汉430030

出　　处：《中国实验血液学杂志》2014年第4期927-931,共5页Journal of Experimental Hematology

基　　金：浙江省宁波市自然科学基金资助(2010A610055)

摘　　要：越来越多的证据显示,肿瘤组织的血管内皮细胞有着与正常血管内皮细胞不同的特征。这些肿瘤血管内皮系统与肿瘤的发生发展密切相关。本研究分离慢性髓系白血病(CML)患者外周血中的循环内皮细胞(circulating endothelial cells,CEC),研究其生物学特性及增殖潜能,并测定其CD133及BCR/ABL融合基因的表达水平,探讨CEC在CML自然病程中的重要作用。用淋巴细胞分离液分离CML患者外周血中的单个核细胞,应用免疫磁珠分选CD45-及CD146+的细胞,在相差显微镜下观察其形态,进行体外培养并绘制其生长曲线;采用免疫荧光染色法测定CEC表面CD31、CD34、VWF及CD133的表达情况;利用FISH技术检测BCR/ABL融合基因的阳性率。结果表明,CML患者外周血中分离的CEC具有典型血管内皮细胞形态,表达血管内皮细胞表面抗原CD31、CD34、VWF,增殖能力明显强于健康对照组,12名CML患者CEC中CD133表达的阳性率为31.29%,与健康对照组相比有显著性差异(P<0.05),BCR/ABL融合基因阳性率为10.77%,与疾病进程呈现正性相关的趋势。结论:CML患者的CEC是肿瘤侵袭发展的重要组成部分,与疾病的传播和扩散有关。Several studies have shown that the tumor endothelial cells are different from the normal tissue endothelial cells. These tumor endothelial cells may contribute to tumor neovasculogenesis. This study was purposed to analyze the biologic features and determine the expression level of CD133 and BCR/ABL fusion gene in circulating endothelial cells （CEC） isolated from peripheral blood of CML patients, as well as to investigate the role of CEC in disease progression. Mononuclear cells were isolated from peripheral blood by density gradient centrifugation; CEC were sorted by MACS and harvested in the endothelial growth medium. The morphologic features of CEC were observed by microscopy, the cell growth rate was calculated by cell counting, and the cells were identified by immunofluorescence staining for the ex- pression of CD31 ,CD34 ,VWF and CD133. The expression of BCR/ABL fusion gene was examined by FISH in 12 CML patients. The results indicated that the isolated CEC displayed the typical cobble-stone morphology. These cells could be identified by the positive immunofluorescence staining for CD31 ∽ CD34 and VWF, and showed more increased prolifera- tive potential as compared to that of healthy donors. It was found that the positive rate of CD133 was 31.29% in CML patients, which was significantly different from that of healthy donors （P 〈 0.05 ）. In 12 CML patients, CEC carded the same chromosome aberration as the leukemia cells （ 10.77% ）. Higher expression level of CD133 and BCR/ABL fusion gene positively correlated with progression of disease. It is concluded that the CEC may participate in invasion and angio- genesis in patients with CML and possibly correlate to the spreading and progression of the disease.

关 键 词：慢性粒细胞白血病 BCR/ABL融合基因 循环内皮细胞 

分 类 号：R733.72[医药卫生—肿瘤]