Objective Tumor stem cells are the subpopulations of stem cell-related markers in tumor cells , which have high proliferative capacity and multi-directional differentiation potential .This paper will explore whether endothelial cells derived from breast cancer stem cells are involved in the angiogenesis of breast cancer . Methods Collected human breast cancer tissue samples , the ex-pression of mutant P 53, CD31, VEGF in tumor vessels were detected by immunohistochemical and immunofluorescence , the amplifica-tion of Her-2 was detected by fluorescence in situ hybridization ;single cell suspension of breast cancer was prepared , using immunomagnetic beads sorting to isolated CD 44+/CD24-/low cells which were cultured in the EGM-2 endothelial cell medium , the ex-pression of endothelial cell surface markers CD 31 and CD105 and the ability of uptake acetylated LDL were detected by flow cytometry .In vitro, three-dimensional plastic culture was carried out , and vessel-like structures were observed. Results The expression of CD31, VEGF and mutant P 53 were observed in paraffin sections of breast cancer tissue samples, and they were arranged along the vessel lu-men or blood vessel sphere .Immunofluorescence showed that CD 31 and DAPI signals were expressed intravascularly in breast cancer tissues.4 samples of Her-2 positive amplification and 9 cases of non-amplification were detected in the 13 samples.The successfully i-solated breast cancer stem cells was performed by immunomagnetic beads sorting .The proportion of CD 44+cells before and after sorting was (7.5±2.6) %and (94.3±4.7)%, respectively, and CD24+cells was (48.2±9.4)%and (4.3±1.6)%, respectively, and the pro-portion of CD24+cells were inversely proportional to CD 24-/low cells.The percentage of CD105+and CD31+cells in mammary gland cells were (4.5±0.9)%and (6.2±1.3)%, respectively, and after continuously cultured for 3 generations in endothelial cells culture system, the percentage of CD105+and CD31+cells were (79.6±9.3)%and (84.1±10.7)%, respectively.It rised significantly.The cells cultured in endothelial cells culture system could phagocytose DiL-Ac-LDL, indicating that the cells had the function of endotheli-al cells.The endothelial cells group showed vascular structure .However, there was no trend of vascular changes in the control group . Conclusion Breast cancer stem cell-derived endothelial cells are able to differentiate into endothelial cells , and participate in the tumor angiogenesis .%目的：肿瘤干细胞是肿瘤细胞中表达干细胞相关标志的亚群，具有高增殖能力和多向分化潜能。文中探讨源于乳腺癌干细胞的内皮细胞是否参与了乳腺癌的血管形成过程，为乳腺癌发病机制提供部分理论基础。方法收集人乳腺癌组织样本，免疫组化法和免疫荧光法检测肿瘤血管中突变型P53、CD31、VEGF的表达，荧光原位杂交方法检测Her-2扩增情况；制备乳腺癌单细胞悬液，采用免疫磁珠分选方法分选出CD44＋／CD24－／low细胞，以培养体系为EGM-2内皮细胞培养基培养，流式细胞检测其内皮细胞表面标志CD31和CD105的表达和摄取乙酰化低密度脂蛋白的能力，体外进行三维胶培养，观察其脉管样结构。结果乳腺癌组织样本的石蜡切片中均可观察到CD31、VEGF、突变型P53的表达，并沿着血管腔或血管球排列；免疫荧光显示乳腺癌组织血管内表达CD31和DAPI信号；在分离成功的乳腺癌干细胞进行免疫磁珠分选，分选前、后细胞中CD44＋细胞比例分别为（7．5±2．6）％、（94．3±4．7）％；分选前、后细胞中CD24＋细胞比例分别为（48．2±9．4）％、（4．3±1．6）％， CD24＋细胞比例越低则CD24－／low细胞比例越高；乳腺球细胞中CD105＋细胞比例为（4．5±0．9）％，CD31＋细胞比例为（6．2±1．3）％；经内皮细胞培养体系持续培养3代后，CD105＋和CD31＋细胞比例明显提高分别为（79．6±9．3）％和（84．1±10．7）％；经内皮细胞培养体系培养后的细胞能吞噬DiL-Ac-LDL，说明该细胞具有内皮细胞类的功能；内皮细胞组可见有脉管样结构形成，而脂肪细胞无脉管样改变的趋势。结论乳腺癌干细胞来源的内皮细胞可分化为血管内皮细胞，在体外培养中可参与肿瘤血管的形成。
展开▼
