乳腺癌干细胞向血管内皮细胞分化及血管形成的实验研究

发布时间：2018-07-04 10:17

本文选题：乳腺癌 + 乳腺癌干细胞　；参考：《西安交通大学学报(医学版)》2017年02期

【摘要】：目的探讨乳腺癌干细胞向血管内皮细胞分化的潜力及参与肿瘤血管形成的作用。方法收集人乳腺癌组织样本,检测肿瘤血管中突变型P53、CD31、VEGF的表达及Her-2扩增;制备乳腺癌单细胞悬液,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,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13例所取样本中检测到Her-2阳性扩增4例,未扩增9例。分离成功的乳腺癌干细胞进行免疫磁珠分选,分选前CD44~+细胞比例[(7.5±2.6)%]明显低于分选后[(94.3±4.7)]%,差异具有统计学意义(P0.05);分选前CD24~+细胞比例[(48.2±9.4)%]明显高于分选后[(4.3±1.6)%],差异具有统计学意义(P0.05)。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~+细胞比例分别为(79.6±9.3)%和(84.1±10.7)%,差异有统计学意义(P0.05)。经内皮细胞培养体系培养后的细胞能吞噬DiL-Ac-LDL,内皮细胞组可见有脉管样结构形成,而对照组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并具有脉管形成的能力,提示乳腺癌干细胞可能参与肿瘤血管的形成。
[Abstract]:Objective to investigate the potential of breast cancer stem cells to differentiate into vascular endothelial cells (VEC) and its role in tumor angiogenesis. Methods Human breast cancer tissue samples were collected to detect the VEGF expression and Her-2 amplification of mutant P53tCD31T in tumor vessels. Single cell suspension of breast cancer was prepared, CD44 ~ / CD24-rlow cells were isolated and cultured on EGM-2 endothelial cell culture medium. The expression of CD31 and CD105 on endothelial cells and the ability of uptake of acetylated low density lipoprotein (LDL) were detected. Results the expression of CD31VEGFand mutant p53 was observed in breast cancer tissues, and the expression of CD31 and DAPI was detected by immunofluorescence. Her-2 positive amplification was detected in 4 cases and no amplification in 9 cases in 13 samples. The percentage of CD44~ cells in breast cancer stem cells before separation [(7.5 卤2.6)%] was significantly lower than that after separation [(94.3 卤4.7)], the difference was statistically significant (P0.05), and the percentage of CD24~ cells before separation [(48.2 卤9.4)%] was significantly higher than that after separation [(4.3 卤1.6)%] (P0.05). The percentage of CD105~ cells in mammary gland bulb cells was (4.5 卤0.9) and (6.2 卤1.3) cells respectively, and the percentages of CD105- and CD31~ cells were (79.6 卤9.3)% and (84.1 卤10.7) after 3 passages of endothelial cell culture system, respectively. The difference was statistically significant (P0.05). The cells cultured in endothelial cell culture system could phagocytosis of DiL-Ac-LDL. Vascular like structure was observed in endothelial cell group, but there was no vascular-like change in control group. Conclusion the endothelial cells derived from breast cancer stem cells can differentiate into vascular endothelial cells and have the ability of vascular formation, suggesting that breast cancer stem cells may be involved in tumor angiogenesis.
【作者单位】：湖北省荆门市第一人民医院甲状腺乳腺外科;湖北省十堰市太和医院湖北医药学院附属医院普外科;
【基金】：荆门市引导性科研项目资助(No.YDKY2016002)~~
【分类号】：R737.9

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