长链非编码 RNA HOTAIRM1 通过调控 miR-873-5p/ZEB2 轴促进胶质瘤细胞的增殖并抑制其凋亡。
Long non-coding RNA HOTAIRM1 promotes proliferation and inhibits apoptosis of glioma cells by regulating the miR-873-5p/ZEB2 axis.
机构信息
Department of Neurosurgery, Zhejiang Tongde Hospital, Hangzhou, Zhejiang 310012, China.
Department of Neurosurgery, Second Affiliated Hospital of Zhejiang University, School of Medicine, Zhejiang University, Hangzhou, Zhejiang 310016, China.
出版信息
Chin Med J (Engl). 2020 Jan 20;133(2):174-182. doi: 10.1097/CM9.0000000000000615.
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.
背景
胶质母细胞瘤是最常见的恶性脑肿瘤之一。传统的胶质母细胞瘤临床治疗并不充分，且其发病和发展的分子机制仍不清楚。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的分子机制。
方法
我们从基因表达综合数据库（GEO）中分析了最失调的 microRNAs，并检测了 20 例胶质母细胞瘤组织和 10 例浙江同德医院收集的正常脑组织中 miR-873-5p 的表达情况。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 的表达，并通过细胞计数试剂盒-8 检测、划痕愈合检测和细胞凋亡分析其表型。此外，我们使用生物信息学分析预测了 miR-873-5p 在胶质母细胞瘤中的上下游基因，并使用荧光素酶报告基因检测和 Western blot 检测在 U87 细胞中验证我们的假设。两组间的差异采用 Student's t 检验分析。多组间的比较采用 Kruskal-Wallis 检验。P 值＜0.05 被认为有统计学意义。
结果
与正常脑组织相比，miR-873-5p 在胶质母细胞瘤组织中表达下调（正常 vs. 肿瘤，0.762±0.231 vs. 0.378±0.114，t=4.540，P＜0.01）。过表达 miR-873-5p 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 则有相反的效果。机制上，我们发现长链非编码 RNA HOTAIRM1 可作为 miR-873-5p 的海绵，激活 U87 细胞中 ZEB2 的表达。
结论
我们揭示了胶质母细胞瘤细胞中 novel HOTAIRM1/miR-873-5p/ZEB2 轴，为胶质母细胞瘤的进展提供了新的见解，并为胶质母细胞瘤的治疗提供了理论基础。
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