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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。
There is growing evidence that vascular endothelial cells of tumor tissue have different characteristics from normal vascular endothelial cells. These tumor vascular endothelial systems are closely related to the occurrence and development of tumors. In this study, circulating endothelial cells (CECs) from peripheral blood of patients with chronic myelogenous leukemia (CML) were isolated and their biological characteristics and proliferation potential were studied. The expression of CD133 and BCR / ABL fusion genes was also examined CEC plays an important role in the natural history of CML. Peripheral blood mononuclear cells from CML patients were isolated by lymphocyte separation fluid. CD45- and CD146 + cells were sorted by immunomagnetic beads. The morphology of the cells was observed under a phase contrast microscope and cultured in vitro. The growth curve was drawn by immunofluorescence staining Method to determine the expression of CD31, CD34, VWF and CD133 on CEC surface. The positive rate of BCR / ABL fusion gene was detected by FISH. The results showed that CEC isolated from peripheral blood of CML patients showed typical morphology of vascular endothelial cells, expressed CD31, CD34 and VWF of vascular endothelial cells, and significantly enhanced the proliferative capacity of CEC patients. The positive rate of CD133 expression in CECs from 12 CML patients (31.29%). The positive rate of BCR / ABL fusion gene was 10.77%, which showed a positive correlation with disease progression. Conclusion: CEC in CML patients is an important part of tumor invasion and development, which is related to the spread and spread of disease.