人脐带间充质干细胞对神经胶质瘤生长的抑制  

Human umbilical cord mesenchymal stem cells inhibit glioma growth

作　　者：梁硕[1,2] 焦红亮[3] 迟连凯[1] 史辛艺[1] 梁阿铭[3] 田毅[3] 韩娇玲[1] 马姗姗[1] 杨波[3] 关方霞[1,4] 

机构地区：[1]郑州大学生物工程系,河南省郑州市450001 [2]河南科技大学第一附属医院生物治疗中心,河南省洛阳市471003 [3]郑州大学第一附属医院神经外科,河南省郑州市450052 [4]河南省医学科学院,河南省郑州市450003

出　　处：《中国组织工程研究》2012年第49期9179-9185,共7页Chinese Journal of Tissue Engineering Research

摘　　要：背景:Wnt信号通路是决定肿瘤发生发展的关键通路。研究表明Dickkopf-1分泌蛋白能阻断Wnt信号向胞内的传递。目的:通过共培养体外观察人脐带间充质干细胞对C6胶质细胞瘤生长的抑制作用及其相关机制。方法:采用不同浓度的人脐带间充质干细胞条件培养液培养C6胶质瘤细胞。分别使用CCK-8和流式细胞仪对细胞增殖和细胞周期进行观察,以Western blotting法检测C6细胞中β-catenin和c-Myc的表达水平。酶联免疫吸附法检测人脐带间充质干细胞分泌Dickkopf-1的水平。利用抗Dickkopf-1的抗体中和人脐带间充质干细胞条件培养液中的Dickkopf-1,检测抗体中和后的效应。结果与结论:人脐带间充质干细胞能够抑制C6细胞的生长,将细胞周期阻滞在G0-G1期。在人脐带间充质干细胞条件培养液的作用下,C6细胞中β-catenin和c-Myc的表达下调。分泌蛋白Dickkopf-1的水平与人脐带间充质干细胞条件培养液的浓度呈正相关。随着条件培养液浓度的增加,人脐带间充质干细胞对C6细胞增殖的抑制效应增强。然而,当抗Dickkopf-1的抗体中和人脐带间充质干细胞条件培养液中的Dickkopf-1后,抑制作用减弱。结果表明,人脐带间充质干细胞通过分泌的一些可溶性因子如Dickkopf-1,能够抑制胶质瘤细胞的生长。BACKGROUND： Wnt signaling pathway is a key to occurrence and development of tumor. Several studies have demonstrated that Dickkopf-1 can block intracellular Wnt signalling transmission. OBJECTIVE： To investigate the inhibitory effect and underlying mechanism of human umbilical cord mesenchyma stem cells on C6 glioma growth by a co-culture system in vitro. METHODS： C6 glioma cells were cultured with human umbilical cord mesenchymal stem cells conditioned medium under different concentrations. Cell Counting Kit-8 and flow cytometry analysis were performed to investigate cell proliferation and cell cycle status. Western blotting was used to detect the expression of β-catenin and c-Myc in C6 cells treated with human umbilicat cord mesenchymal stem cells conditioned medium. Enzyme-linked immunosorbent assay was adopted to examine the level of Dickkopf-1 secreted by human umbilical cord mesenchymal stem cells. Dickkopf-1 in human umbilical cord mesenchymal stem cells conditioned medium was neutralized by anti-Dickkopf-1 antibody. The effects of antibody neutralization were also determined by the above methods. RESULTS AND CONCLUSION： Human umbilical cord mesenchymal stem cells could inhibit C6 cell expansion and arrest the cell cycle in G0-G1 phase. The expression levels ofβ-catenin and c-Myc were down-regulated in C6 cells after treatment with human umbilical cord mesenchymal stem cells conditioned medium. The levels of secreted protein Dickkopf-1 were positively correlated with concentrations of human umbilical cord mesenchymal stem cells conditioned medium. The inhibitory effect of human umbilical cord mesenchymal stem cells on C6 cell proliferation was enhanced as the concentration of Dickkopf-1 in human umbilical cord mesenchymal stem cells conditioned medium increased. When Dickkopf-1 was neutralized by anti-Dickkopf-1 antibody, the suppressing effect was attenuated. It demonstrated that human umbilical cord mesenchvmal stem cells could inhibit qlioma cell growth via secreting the soluble factors, such

关 键 词：人脐带 间充质干细胞 胶质瘤 抑制 DICKKOPF-1 干细胞 

分 类 号：R394.2[医药卫生—医学遗传学]