CAJ | 학술논문

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.
배경:Wnt신호통로시결정종류발생발전적관건통로.연구표명Dickkopf-1분비단백능조단Wnt신호향포내적전체.목적:통과공배양체외관찰인제대간충질간세포대C6효질세포류생장적억제작용급기상관궤제.방법:채용불동농도적인제대간충질간세포조건배양액배양C6효질류세포.분별사용CCK-8화류식세포의대세포증식화세포주기진행관찰,이Western blotting법검측C6세포중β-catenin화c-Myc적표체수평.매련면역흡부법검측인제대간충질간세포분비Dickkopf-1적수평.이용항Dickkopf-1적항체중화인제대간충질간세포조건배양액중적Dickkopf-1,검측항체중화후적효응.결과여결론:인제대간충질간세포능구억제C6세포적생장,장세포주기조체재G0-G1기.재인제대간충질간세포조건배양액적작용하,C6세포중β-catenin화c-Myc적표체하조.분비단백Dickkopf-1적수평여인제대간충질간세포조건배양액적농도정정상관.수착조건배양액농도적증가,인제대간충질간세포대C6세포증식적억제효응증강.연이,당항Dickkopf-1적항체중화인제대간충질간세포조건배양액중적Dickkopf-1후,억제작용감약.결과표명,인제대간충질간세포통과분비적일사가용성인자여Dickkopf-1,능구억제효질류세포적생장.
BACKGROUND:Wnt signaling pathway is a key to occurrence and development of tumor. Several studies have demonstrated that Dickkopf-1 can block intracel ular Wnt signal ing transmission. OBJECTIVE:To investigate the inhibitory effect and underlying mechanism of human umbilical cord mesenchymal stem cel s on C6 glioma growth by a co-culture system in vitro. METHODS:C6 glioma cel s were cultured with human umbilical cord mesenchymal stem cel s conditioned medium under different concentrations. Cel Counting Kit-8 and flow cytometry analysis were performed to investigate cel proliferation and cel cycle status. Western blotting was used to detect the expression ofβ-catenin and c-Myc in C6 cel s treated with human umbilical cord mesenchymal stem cel s conditioned medium. Enzyme-linked immunosorbent assay was adopted to examine the level of Dickkopf-1 secreted by human umbilical cord mesenchymal stem cel s. Dickkopf-1 in human umbilical cord mesenchymal stem cel s 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 cel s could inhibit C6 cel expansion and arrest the cel cycle in G0-G1 phase. The expression levels ofβ-catenin and c-Myc were down-regulated in C6 cel s after treatment with human umbilical cord mesenchymal stem cel s conditioned medium. The levels of secreted protein Dickkopf-1 were positively correlated with concentrations of human umbilical cord mesenchymal stem cel s conditioned medium. The inhibitory effect of human umbilical cord mesenchymal stem cel s on C6 cel proliferation was enhanced as the concentration of Dickkopf-1 in human umbilical cord mesenchymal stem cel s conditioned medium increased. When Dickkopf-1 was neutralized by anti-Dickkopf-1 antibody, the suppressing effect was attenuated. It demonstrated that human umbilical cord mesenchymal stem cel s could inhibit glioma cel growth via secreting the soluble factors, such as Dickkopf-1.