Medical School of Southeast University, Nanjing, China.
Department of Neurosurgery, Affiliated People's Hospital of Jiangsu University, Zhenjiang, China.
J Cell Mol Med. 2020 Oct;24(20):11755-11767. doi: 10.1111/jcmm.15788. Epub 2020 Sep 11.
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.
我们的目的是确定长链非编码 RNA（lncRNA）HOXA-AS3 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 微阵列分析来确定与 GBM 相关的 lncRNA 表达谱。Qrt-PCR 用于调查长非编码 RNA（lncRNA）HOXA-AS3 和靶基因的表达水平。双荧光素酶报告基因实验用于研究 lncRNA HOXA-AS3、靶基因和 miRNA 的相互作用。Western blot 分析用于检测 USP3 和上皮间质转化（EMT）基因的表达。MTT 分析、Transwell 分析和划痕愈合分析分别用于分析 lncRNA HOXA-AS3 对 GBM 细胞活力、迁移和侵袭的影响。我们的结果表明，lncRNA HOXA-AS3 在 GBM 细胞中显著上调，并能促进 GBM 细胞在体外和体内的增殖、迁移和侵袭。HOXA-AS 与 glioma 患者的不良生存预后相关。双荧光素酶报告基因实验还揭示了 lncRNA HOXA-AS3 通过上调 USP3 表达来充当 mir-455-5p 海绵，从而促进 GBM 的进展。Western blot 分析表明，lncRNA HOXA-AS3 可上调 GBM 中 EMT 相关基因的表达。实验表明，mir-455-5p 可以挽救 lncRNA HOXA-AS3 对细胞增殖和侵袭的影响。新鉴定的 HOXA-AS3/mir-455-5p/USP3 通路为理解 lncRNA HOXA-AS3 在胶质母细胞瘤中作用的关键机制提供了重要线索。
