内皮细胞对髓母细胞瘤干细胞特性的影响  

Endothelial cells regulates medulloblastom stem-like cells characteristics by Notch pathway activation

作　　者：王洪新[1] 王玉社[1,2] 王勇 WANG Hongxin;WANG Yushe;WANG Yong(Henan Provincial People's Hospital/Henan University People's Hospital,Zhengzhou 450003,China;Henan Provincial Cerebrovascular Disease Hospital,Zhengzhou 450003,China)

机构地区：[1]河南省人民医院河南大学人民医院,河南郑州450003 [2]河南省脑血管病医院,河南郑州450003

出　　处：《中国实用神经疾病杂志》2020年第22期1933-1938,共6页Chinese Journal of Practical Nervous Diseases

基　　金：河南省卫健委省部共建青年项目(编号:SB201902027)。

摘　　要：目的探讨内皮细胞对髓母细胞瘤干细胞特性的影响及机制。方法实验分为3组,A组D341Med(D341髓母细胞瘤系);B组D341Med+HBMECsD341髓母细胞瘤系和人脑微血管内皮细胞(human brain microvasculature endothelial cells,HBMECs)共培养;C组D341Med+HBMECs+DAPTD341髓母细胞瘤系和人脑微血管内皮细胞以及γ-分泌酶抑制剂(DAPT)共培养,成球试验和流式细胞仪检测CD133+细胞,观察对肿瘤干细胞的影响。采用实时定量PCR和Western印迹法检测Notch(Notch 2、Jagded 1、Hes-1和Hey-2)mRNA和蛋白的表达,Transwell侵袭试验检测内皮细胞对肿瘤侵袭能力的影响,裸鼠成瘤试验观察肿瘤细胞的生长。结果与A组相比,B组成球数目和CD133+细胞增加(P<0.01),Notch相关mRNA和蛋白表达水平增加(P<0.05);Transwell侵袭试验显示,穿膜细胞数明显增高(P<0.05);裸鼠的肿瘤体积增大(P<0.05)。与B组相比,C组干细胞标记CD133+细胞急剧下降(P<0.05),Notch相关mRNA和蛋白表达水平下降(P<0.05);Transwell侵袭试验显示,穿膜细胞数明显下降(P<0.05);裸鼠的肿瘤体积缩小(P<0.05)。结论内皮细胞可能通过激活Notch通路调控髓母细胞瘤干细胞特性。Objective To investigate the effects of human brain microvasculature endothelial cells on the stem-like cells characteristics of medulloblastom and to explore its mechanism.Methods The samples were divided into three groups,group AD341Med;group BD341Med cells and HBMECs were co-cultivated;group CD341Med cells and HBMECs and DAPT were co-cultivated.The proportion of stem-like cells was investigated by sphere-forming assay and flow cytometry.Notch(Notch2,Jagged 1,Hes-1,and Hey-2)were quantified by quantitative real-time PCR and Western blot.Transwell invasion assay was used to detect invasion ability.Mice xenograft experiments were used to observe the tumorigenesis in vivo.Results Compared with group A,the sphere-forming rate and cell ratio of stem-like cells were significantly increased in group B(P<0.01).The expression of genes and protein of Notch were obviously upregulated in group B(P<0.05).Transwell invasion assay revealed that the number of migrating cells in group B were significantly larger than group A(P<0.05).Tumor size and volume in group B enlarged compared with group A(P<0.05).Compared with group B,the percentage of CD133+were significantly decreased in group C(P<0.05).The expression of genes and protein of Notch were obviously downregulated in group C(P<0.05).Transwell invasion assay revealed that the number of migrating cells in group C were significantly lower than group B(P<0.05).Tumor size and volume in group C shrunk compared with group B(P<0.05).Conclusion Human brain microvasculature endothelial cells regulates medulloblastom stem-like cells characteristics probably by activating Notch signal pathway.

关 键 词：髓母细胞瘤 干细胞 内皮细胞 NOTCH通路 Transwell侵袭试验 

分 类 号：R-332[医药卫生]