背景:胶质母细胞瘤是最常见的恶性脑肿瘤之一。胶质母细胞瘤的常规临床治疗不充分,这种疾病的发生和发展的分子机制尚不清楚。本研究旨在探讨miR-873a-5p在胶质母细胞瘤中的表达和功能及其相关分子机制。
方法:我们分析了来自基因表达Omnibus(GEO)数据库中最失调的microRNAs,并与浙江同德医院收集的10个正常脑组织相比,检测了20个胶质母细胞瘤组织中miR-873-5p的表达。然后,我们在U87胶质母细胞瘤细胞系中过表达或抑制miR-873-5p表达,并使用细胞计数试剂盒-8测定分析表型,伤口愈合试验,和凋亡。此外,我们使用生物信息学分析在胶质母细胞瘤中预测miR-873-5p的上游和下游基因,并使用荧光素酶报告基因分析和Western印迹分析在U87细胞中检验我们的假设.采用Studentt检验分析两组间的差异。使用Kruskal-Wallis检验对多组进行比较。P<0.05被认为是显著的。
结果:与正常脑组织相比,胶质母细胞瘤组织中的miR-873-5p下调(正常与肿瘤,0.762±0.231vs.0.378±0.114,t=4.540,P<0.01)。miR-873-5p过表达抑制细胞生长(t=6.095,P<0.01)和迁移(t=3.142,P<0.01),促进细胞凋亡(t=4.861,P<0.01);而抑制miR-873-5p具有相反的作用。机械上,发现长链非编码RNAHOTAIRM1充当miR-873-5p的海绵,激活U87细胞中ZEB2的表达.
结论:我们在胶质母细胞瘤细胞中发现了一个新的HOTAIRM1/miR-873-5p/ZEB2轴,为胶质母细胞瘤的进展提供新的见解,为胶质母细胞瘤的治疗提供理论依据。
BACKGROUND: Glioblastoma is one of the most common malignant brain tumors. Conventional clinical treatment of glioblastoma is not sufficient, and the molecular mechanism underlying the initiation and development of this disease remains unclear. Our study aimed to explore the expression and function of miR-873a-5p in glioblastoma and related molecular mechanism.
METHODS: We analyzed the most dysregulated microRNAs from the Gene Expression Omnibus (GEO) database and examined the expression of miR-873-5p in 20 glioblastoma tissues compared with ten normal brain tissues collected in the Zhejiang Tongde Hospital. We then overexpressed or inhibited miR-873-5p expression in U87 glioblastoma cell lines and analyzed the phenotype using the cell counting kit-8 assay, wound healing assay, and apoptosis. In addition, we predicted upstream and downstream genes of miR-873-5p in glioblastoma using bioinformatics analysis and tested our hypothesis in U87 cells using the luciferase reporter gene assay and Western blotting assay. The differences between two groups were analyzed by Student\'s t test. The Kruskal-Wallis test was used for the comparison of multiple groups. A P < 0.05 was considered to be significant.
RESULTS: The miR-873-5p was downregulated in glioblastoma tissues compared with that in normal brain tissues (normal vs. tumor, 0.762 ± 0.231 vs. 0.378 ± 0.114, t = 4.540, P < 0.01). Overexpression of miR-873-5p inhibited cell growth (t = 6.095, P < 0.01) and migration (t = 3.142, P < 0.01) and promoted cell apoptosis (t = 4.861, P < 0.01), while inhibition of miR-873-5p had the opposite effect. Mechanistically, the long non-coding RNA HOTAIRM1 was found to act as a sponge of miR-873-5p to activate ZEB2 expression in U87 cells.
CONCLUSIONS: We uncovered a novel HOTAIRM1/miR-873-5p/ZEB2 axis in glioblastoma cells, providing new insight into glioblastoma progression and a theoretical basis for the treatment of glioblastoma.