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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 in situ 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 in vivo. 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：
胶质瘤是一种预后不良的普通恶性肿瘤。长链非编码RNA（lncRNA）与肿瘤的起始和过程有关。对GEPIA数据库的调查显示，与正常脑组织相比，神经胶质瘤组织中的长链非编码RNAWEE2反义RNA1（WEE2-AS1）上调，定量实时聚合酶链反应（qRT-PCR）验证显示，WEE2-AS1表达与数据库预测一致。荧光原位杂交（FISH）分析显示，WEE2-AS1主要位于细胞质中。克隆形成实验和EDU实验检测细胞增殖能力,用Transwell法检测细胞的迁移和侵袭能力,免疫印迹法和免疫荧光法测定TPM3蛋白水平。功能实验表明，WEE2-AS1的下调阻碍了细胞增殖，迁移,和侵袭神经胶质瘤细胞系。此外，WEE2-AS1的下调抑制了体内肿瘤的生长。生物信息学预测和整合实验表明，WEE2-AS1通过构建miR-29b-2-5p促进原肌球蛋白3（TPM3）的表达。进行双荧光素酶报告基因测定以揭示WEE2-AS1和miR-29b-2-5p的结合以及miR-29b-2-5p和TPM3的结合。此外,一系列的拯救实验表明，WEE2-AS1促进增殖，迁移,并通过靶向miR-29b-2-5p来调节TPM3的表达。最终,这项研究的结果表明,WEE2-AS1在神经胶质瘤中起致癌作用,并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.
