The Fourth Department of Neurosurgery, Tangshan Gongren Hospital, Tangshan, Hebei, China.
The Fourth Department of Neurology, Tangshan Gongren Hospital, Tangshan, Hebei, China.
Biosci Rep. 2020 Aug 28;40(8). doi: 10.1042/BSR20200487.
The role of lncRNAs in tumor has been widely concerned. The present study took HAS2-AS1 (the antisense RNA 1 of HAS2) as a starting point to explore its expression in glioma and its role in the process of migration and invasion, providing a strong theoretical basis for mining potential therapeutic targets of glioma.
Clinical data of glioma were obtained from The Cancer Genome Atlas (TCGA) database and differentially expressed lncRNAs were analyzed by edgeR. The hTFtarget database was used to predict the upstream transcription factors of HAS2-AS1 and the JASPAR website was used to predict the binding sites of human upstream transcription factor 1 (USF1) and HAS2-AS1. qRT-PCR was used to detect the expressions of HAS2-AS1 and USF1 in glioma tissues and cell lines. The effects of silencing HAS2-AS1 on the migration and invasion of cancer cells were verified by wound healing and Transwell invasion assays. The chromatin immunoprecipitation (ChIP) and dual luciferase reporter assays were applied to demonstrate the binding of USF1 and HAS2-AS1 promoter region. Western blot was used to detect the expressions of epithelial-mesenchymal transition (EMT)-related proteins.
HAS2-AS1 was highly expressed in glioma tissues and cells, and was significantly associated with poor prognosis. Silencing HAS2-AS1 expression inhibited glioma cell migration, invasion and EMT. USF1 was highly expressed in glioma and positively correlated with HAS2-AS1. The transcription of HAS2-AS1 was activated by USF1 via binding to HAS2-AS1 promoter region, consequently potentiating the invasion and migration abilities of glioma cells.
These results suggested that the transcription factor USF1 induced up-regulation of lncRNA HAS2-AS1 and promoted glioma cell invasion and migration.
长链非编码 RNA（lncRNA）在肿瘤中的作用已受到广泛关注。本研究以 HAS2-AS1（HAS2 的反义 RNA1）为起点，探讨其在脑胶质瘤中的表达及其在迁移和侵袭过程中的作用，为挖掘脑胶质瘤潜在治疗靶点提供了强有力的理论依据。
从癌症基因组图谱（TCGA）数据库中获取脑胶质瘤的临床数据，采用 edgeR 分析差异表达的 lncRNA。使用 hTFtarget 数据库预测 HAS2-AS1 的上游转录因子，使用 JASPAR 网站预测人上游转录因子 1（USF1）和 HAS2-AS1 的结合位点。qRT-PCR 检测脑胶质瘤组织和细胞系中 HAS2-AS1 和 USF1 的表达。通过划痕愈合和 Transwell 侵袭实验验证沉默 HAS2-AS1 对癌细胞迁移和侵袭的影响。采用染色质免疫沉淀（ChIP）和双荧光素酶报告基因实验证实 USF1 与 HAS2-AS1 启动子区的结合。Western blot 检测上皮间质转化（EMT）相关蛋白的表达。
HAS2-AS1 在脑胶质瘤组织和细胞中高表达，与预后不良显著相关。沉默 HAS2-AS1 表达抑制脑胶质瘤细胞迁移、侵袭和 EMT。USF1 在脑胶质瘤中高表达，与 HAS2-AS1 呈正相关。USF1 通过结合 HAS2-AS1 启动子区激活 HAS2-AS1 的转录，从而增强脑胶质瘤细胞的侵袭和迁移能力。
这些结果表明转录因子 USF1 诱导 lncRNA HAS2-AS1 的上调，并促进脑胶质瘤细胞的侵袭和迁移。
