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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 the key path that determines the development of tumor. Studies have shown that Dickkopf-1 secreted protein can block the Wnt signal to the intracellular transmission. OBJECTIVE: To observe the inhibitory effect of human umbilical cord mesenchymal stem cells on the growth of C6 glioma by co-culture and its related mechanism. Methods: C6 glioma cells were cultured with different concentrations of human umbilical cord mesenchymal stem cell conditioned medium. Cell proliferation and cell cycle were observed by CCK-8 and flow cytometry respectively. The expression of β-catenin and c-Myc in C6 cells was detected by Western blotting. The level of Dickkopf-1 secreted by human umbilical cord mesenchymal stem cells was detected by enzyme-linked immunosorbent assay. Dickkopf-1 in human umbilical cord mesenchymal stem cell conditioned medium was neutralized with an anti-Dickkopf-1 antibody to examine the effect of antibody neutralization. RESULTS AND CONCLUSION: Human umbilical cord mesenchymal stem cells can inhibit the growth of C6 cells and block the cell cycle in G0-G1 phase. Under the condition of human umbilical cord mesenchymal stem cell conditioned medium, the expression of β-catenin and c-Myc in C6 cells was down-regulated. The level of secretory protein Dickkopf-1 was positively correlated with the concentration of human umbilical cord mesenchymal stem cell conditioned medium. With the increasing concentration of conditioned medium, human umbilical cord mesenchymal stem cells inhibit the proliferation of C6 cells. However, the anti-Dickkopf-1 antibody neutralizes Dickkopf-1 in human umbilical cord mesenchymal stem cell conditioned medium. The results showed that human umbilical cord mesenchymal stem cells can inhibit the growth of glioma cells by secreting some soluble factors such as Dickkopf-1.