摘要
目的研究mi R-144对人U87来源的胶质瘤干细胞(glioma stem cells,GSCs)生物学行为的影响。方法以干细培养液培养人U87胶质瘤细胞2周后,应用免疫荧光法检测细胞球的Nestin及CD133的表达,同时细胞球在含有10%胎牛血清培养基中分化培养2周,应用免疫荧光法检测GFAP和tubulin III的表达。过表达或抑制mi R-144的表达,应用CCK8法检测GSCs的细胞活力,应用FITC-PI双染色方法检测细胞凋亡,应用Transwell小室方法检测细胞迁移及侵袭能力。结果细胞球表达Nestin及CD133。细胞球分化培养2周后,细胞表达GFAP及tubulin III。与non-GSCs相比,在GSCs中mi R-144的表达明显降低。与阴性对照组相比,过表达mi R-144显著抑制了GSCs的细胞活力和细胞迁移及侵袭能力,诱导了GSCs的凋亡。而抑制mi R-144的表达显著提高了GSCs的细胞活力和细胞迁移及侵袭能力,抑制了GSCs的凋亡。结论 Mi R-144能够显著抑制人U87来源的GSCs的细胞活力及细胞迁移及侵袭能力,诱导GSCs的凋亡。
Objective To investigate the effects of mi R-144 on the biological behaviors of glioma stem cells（GSCs） derived from human U87 glioma cells. Methods Human U87 glioma cells were cultured in the stem cell medium for 2 weeks, the expression of Nestin and CD133 in cell spheres was assessed by fluorescence microscopy, meanwhile after cell spheres were cultured in medium with 10% fetal bovine serum for 2 weeks, the expression of GFAP and tubulin III was determined by fluorescence microscopy. After over-expression or inhibition of mi R-144, CCK8 assay was used to detect the cell viability of GSCs, FITC-PI staining was used to detect cell apoptosis of GSCs, and Transwell assay was used to detect the cell migration and invasion of GSCs. Results GSCs spheres expressed Nestin and CD133 positively. After cell spheres differentiated 2 weeks, the cells expressed GFAP and tubulin III. Compared with non-GSCs, mi R-144 expression level was significantly downregulated in GSCs. Compared with negative control, overexpression of mi R-144 significantly inhibited the viability and migration and invasion of GSCs, and induced GSCs apoptosis. However, inhibition of mi R-144 significantly upregulated the viability and migration and invasion of GSCs, and inhibited GSCs apoptosis. Conclusion Mi R-144 could remarkably inhibit the viability and migration and invasion, and induce the apoptosis of human GSCs.
出处
《解剖科学进展》
CAS
2015年第2期121-126,共6页
Progress of Anatomical Sciences
基金
国家自然科学基金资助项目(No.81172197
81372484)
沈阳科学技术计划资助项目(F13-220-9-15)
