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LncRNA WEE2-AS1 敲低通过调控 miR-29b-2-5p/TPM3 轴抑制胶质瘤细胞的增殖、迁移和侵袭。

LncRNA WEE2-AS1 knockdown inhibits the proliferation, migration and invasion of glioma cells via regulating miR-29b-2-5p/TPM3 axis.

机构信息

Nanjing Medical University, Nanjing, 210000, China.

Department of Neurosurgery, Nanjing First Hospital, Nanjing Medical University, Nanjing, 210006, China.

出版信息

Oncol Res. 2022 Jul 13;29(2):105-117. doi: 10.32604/or.2022.03536. eCollection 2021.

Abstract

Glioma is a general malignant tumor with a dismal prognosis. Long noncoding RNAs (lncRNAs) have been implicated in the initiation and processes of tumors. An investigation of the GEPIA database revealed that long noncoding RNA WEE2 antisense RNA 1 (WEE2-AS1) is upregulated in glioma tissues compared to normal brain tissues, and validation with quantitative real-time polymerase chain reaction (qRT-PCR) revealed that WEE2-AS1 expression was consistent with the database prediction. Fluorescence hybridization (FISH) assays revealed that WEE2-AS1 was localized primarily in the cytoplasm. Clone formation experiment and EDU assay were used to detect cell proliferation ability, and Transwell assay was used to detect cell migration and invasion ability, Western-blot assay and immunofluorescence were used to determine TPM3 protein level. Functional experiments revealed that the downregulation of WEE2-AS1 impeded cell proliferation, migration, and invasion in glioma cell lines. Furthermore, downregulation of WEE2-AS1 suppressed tumor growth . Bioinformatics predictions and integrated experiments indicated that WEE2-AS1 promoted tropomyosin 3 (TPM3) expression by sponging miR-29b-2-5p. A dual-luciferase reporter assay was conducted to uncover the binding of WEE2-AS1 and miR-29b-2-5p and that of miR-29b-2-5p and TPM3. Additionally, a series of rescue assays showed that WEE2-AS1 promotes proliferation, migration, and invasion by targeting miR-29b-2-5p to regulate TPM3 expression. Ultimately, the results of this study indicate that WEE2-AS1 plays an oncogenic role in glioma and may promote further investigations of the diagnostic and prognostic value of WEE2-AS1 in glioma.

摘要

神经胶质瘤是一种预后不良的常见恶性肿瘤。长链非编码 RNA(lncRNA)被认为参与了肿瘤的发生和发展过程。通过对 GEPIA 数据库的研究发现,长链非编码 RNA WEE2 反义 RNA 1(WEE2-AS1)在神经胶质瘤组织中的表达高于正常脑组织,通过实时定量聚合酶链反应(qRT-PCR)验证发现 WEE2-AS1 的表达与数据库预测一致。荧光原位杂交(FISH)实验表明 WEE2-AS1 主要定位于细胞质。克隆形成实验和 EDU 检测用于检测细胞增殖能力,Transwell 检测用于检测细胞迁移和侵袭能力,Western-blot 检测和免疫荧光检测用于检测 TPM3 蛋白水平。功能实验表明下调 WEE2-AS1 可抑制神经胶质瘤细胞系的增殖、迁移和侵袭。此外,下调 WEE2-AS1 可抑制肿瘤生长。生物信息学预测和综合实验表明,WEE2-AS1 通过海绵吸附 miR-29b-2-5p 促进原肌球蛋白 3(TPM3)的表达。双荧光素酶报告实验揭示了 WEE2-AS1 与 miR-29b-2-5p 的结合以及 miR-29b-2-5p 与 TPM3 的结合。此外,一系列的挽救实验表明,WEE2-AS1 通过靶向 miR-29b-2-5p 调节 TPM3 表达来促进增殖、迁移和侵袭。最终,本研究结果表明,WEE2-AS1 在神经胶质瘤中发挥致癌作用,可能促进进一步研究 WEE2-AS1 在神经胶质瘤中的诊断和预后价值。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2b4d/10207975/62ba233c6774/OncolRes-29-3536-f001.jpg

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