LncRNA HOXA-AS3 promotes the malignancy of glioblastoma through regulating miR-455-5p/USP3 axis.
LncRNA HOXA-AS3 promotes the malignancy of glioblastoma through regulating miR-455-5p/USP3 axis.
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LncRNA HOXA-AS 3通过调控miR-455- 5 p/USP 3轴促进胶质母细胞瘤恶性化。
DOI：
10.1111/jcmm.15788
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发表时间：
2020-10
影响因子：
5.3
通讯作者：
Chen L
中科院分区：
文献类型：
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作者：
Chen W;Li Q;Zhang G;Wang H;Zhu Z;Chen L
我们的目标是确定lncRNA HOXA-AS 3调节多形性胶质母细胞瘤（GBM）生物学行为的分子机制。我们使用lncRNA微阵列测定来鉴定GBM相关的lncRNA表达谱。使用Qrt-PCR来调查长非编码RNA（lncRNA）HOXA-AS 3和靶基因的表达水平。使用双荧光素酶报告基因测定来研究lncRNA HOXA-AS 3、靶基因和miRNA的相互作用。Western blot分析用于检查USP 3和上皮间质转化（EMT）基因的表达。分别采用MTT法、transwell法和伤口愈合法分析lncRNA HOXA-AS 3对GBM细胞活力、迁移率和侵袭力的影响。我们的研究结果表明，lncRNA HOXA-AS 3在GBM细胞中显著上调，并可促进GBM细胞在体外和体内的增殖、侵袭和迁移。发现HOXA-AS与胶质瘤患者的不良生存预后相关。双荧光素酶报告基因测定还显示，lncRNA HOXA-AS 3通过上调USP 3表达来促进GBM进展，从而充当mir-455 - 5 p海绵。Western blot分析显示lncRNA HOXA-AS 3可上调GBM中EMT相关基因的表达。实验表明，mir-455 - 5 p可以挽救lncRNA HOXA-AS 3对细胞增殖和侵袭的影响。新发现的HOXA-AS 3/mir-455 - 5 p/USP 3通路为理解lncRNA HOXA-AS 3在胶质母细胞瘤中作用的关键机制提供了重要线索。
Our objective was to determine the molecular mechanisms by which lncRNA HOXA‐AS3 regulates the biological behaviour of glioblastoma multiforme (GBM). We used an lncRNA microarray assay to identify GBM‐related lncRNA expression profiles. Qrt‐PCR was used to survey the levels of expression of long non‐coding RNA (lncRNA) HOXA‐AS3 and the target gene. Dual‐luciferase reporter assays were used to investigate the interaction of lncRNA HOXA‐AS3, the target gene and miRNA. Western blot analysis was used to examine the expression of USP3 and epithelial‐mesenchymal transition (EMT) genes. The MTT assay, transwell assay and wound healing assay were used to analyse the effects of lncRNA HOXA‐AS3 on GBM cell viability, mobility and invasiveness, respectively. Our results showed that lncRNA HOXA‐AS3 was significantly up‐regulated in GBM cells and could promote GBM cell proliferation, invasion and migration in vitro and in vivo. HOXA‐AS was found to be associated with poor survival prognosis in glioma patients. The dual‐luciferase reporter assay also revealed that lncRNA HOXA‐AS3 acts as a mir‐455‐5p sponge by up‐regulating USP3 expression to promote GBM progression. Western blot analysis showed that lncRNA HOXA‐AS3 could up‐regulate EMT‐related gene expression in GBM. Experiments showed mir‐455‐5p could rescue the effect of lncRNA HOXA‐AS3 on cell proliferation and invasion. The newly identified HOXA‐AS3/mir‐455‐5p/USP3 pathway offers important clues to understanding the key mechanisms underlying the action of lncRNA HOXA‐AS3 in glioblastoma.