MiR-218-5p抑制A2B5^+/CD133~–亚群胶质瘤干细胞的干性维持及侵袭性  

MiR-218-5p Inhibited the Stemness Maintaining and Invasion of A2B5^+/CD133~– Subgroup Human Glioma Stem Cells

作　　者：吴至武 王杭州[2] 李炎炎[1] 李学涛[1] 孙婷[1] 陈桂林[1] 谢学顺[1] 周幽心[1] 杜子威[1] 

机构地区：[1]苏州大学附属第一医院神经外科暨脑神经研究室,苏州215006 [2]苏州大学附属儿童医院,苏州215003

出　　处：《中国细胞生物学学报》2015年第4期469-476,共8页Chinese Journal of Cell Biology

基　　金：国家自然科学基金(批准号:81372689);江苏省卫生厅重点科研项目(批准号:K201106)资助的课题~~

摘　　要：该文目的为探讨miR-218-5p对表达A2B5+/CD133–的胶质瘤干细胞(glioma stem cells,GSCs)亚群干性维持及侵袭性的影响。实时荧光定量PCR(q RT-PCR)检测人脑胶质瘤组织及细胞株中miR-218-5p的表达;上、下调miR-218-5p慢病毒转染表达A2B5+/CD133–的胶质瘤干细胞SHG-139s,二次球体形成实验、免疫荧光及q RT-PCR检测SHG-139s的标志物A2B5、CD133、Nestin、PLAGL2、ALDH1及SOX2的表达变化;Transwell实验、q RT-PCR及免疫荧光检测SHG-139s基质金属蛋白酶9(matrix metalloproteinase 9,MMP9)的表达变化。结果显示,miR-218-5p在胶质瘤中低表达,上调miR-218-5p可显著降低SHG-139s成球率,同时明显降低SHG-139s中上述干细胞标志物的表达;Transwell实验显示,上调miR-218-5p能显著抑制SHG-139s的侵袭能力,使其MMP9表达受到明显抑制。因此,miR-218-5p可能作为抑癌基因在胶质瘤中发挥作用,高表达的miR-218-5p可抑制SHG-139s的干性维持及其侵袭能力。The article was aimed at detecting the expression of miR-218-5p in human gliomas, and exploring its influence in stemness maintaining and invasion of A2B5＋/CD133– human glioma stem cells（SHG-139s）. q RT-PCR was used to detect miR-218-5p expression in noncancerous brain tissue, human glioma tissue, human glioma cell lines and human glioma stem cell lines. Lentivirus vectors encoding miR-218-5p and antimiR-218-5p were constructed and stably transfected into A2B5＋/CD133– SHG-139 s cells. Neurosphere formation assay, Transwell assay, immunofluorescence and q RT-PCR analysis were used to explore the role of miR-218-5p in SHG-139 s. q RT-PCR analysis showed that miR-218-5p expression was lower in human glioma tissues and cells than in noncancerous brain tissue and normal human astrocyte cells, and lower in A2B5＋/CD133–（SHG-139s） cells than in CD133＋ cells（SU2, U87s） cells. Immunofluorescence staining and q RT-PCR showed that miR-218-5p reduced the expressions of stem cell markers（A2B5, nestin, PLAGL2, ALDH1, Sox2）, compared to controls; However, q RT-PCR analysis showed that upregulated miR-218-5p expression led to a decline in CD133 expression, although this result was not confirmed by immunofluorescence staining. Mi R-218-5p reduced SHG-139 s cell invasiveness in Transwell assay and reduced MMP9 expression detected by q RT-PCR and immunofluorescence analysis. Mi R-218-5p is implicated as a tumor suppressor gene in glioma that functions by upregulating miR-218-5p expression, which inhibits the stem cell and invasive properties of SHG-139 s.

关 键 词：miR-218-5p A2B5+/CD133–胶质瘤干细胞 干性维持 侵袭 

分 类 号：R739.4[医药卫生—肿瘤]