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长链非编码 RNA H19 通过靶向 miR-342 调控 Wnt5a/β-连环蛋白通路促进胶质瘤细胞的增殖、迁移和血管生成。

LncRNA H19 Promotes Cell Proliferation, Migration, and Angiogenesis of Glioma by Regulating Wnt5a/β-Catenin Pathway via Targeting miR-342.

机构信息

Department of Oncology, Xiangya Hospital, Central South University, No. 87 Xiangya Road, Kaifu District, Changsha, 410008, Hunan Province, People's Republic of China.

Department of Internal Medicine, Qidong Hospital of Traditional Chinese Medicine, Hengyang, 421600, Hunan Province, People's Republic of China.

出版信息

Cell Mol Neurobiol. 2022 May;42(4):1065-1077. doi: 10.1007/s10571-020-00995-z. Epub 2020 Nov 7.

Abstract

Glioma is the most common malignant brain tumor and long non-coding RNAs (lncRNAs) have been reported to play an important role in the growth and angiogenesis of glioma. However, the potential mechanisms of lncRNA H19 in glioma remain unclear. In the present study, the effects of lncRNA H19 on glioma cell proliferation, migration, and angiogenesis were evaluated. The expression levels of H19, miR-342, and Wnt5a in glioma tissues and cells were detected by RT-qPCR or Western blotting. Dual luciferase reporter assay confirmed the interaction between H19, miR-342, and Wnt5a. Cell proliferation, migration, and angiogenesis were analyzed by colony formation, transwell, and tube formation assays, respectively. IHC was performed to test the angiogenesis-related factor CD31. H19 and Wnt5a expression were remarkably upregulated in glioma tissues and cells, whereas miR-342 expression was downregulated. Moreover, functional analysis confirmed that knockdown of H19 or overexpression of miR-342 suppressed glioma cell proliferation, migration, and angiogenesis in vitro. Besides, H19 was found to directly target miR-342 to promote Wnt5a expression and activate β-catenin pathway in glioma cells. Moreover, suppression of miR-342 or overexpression of Wnt5a reversed the inhibitory effect of sh-H19 on glioma growth and metastasis. Additionally, we verified that H19 promoted glioma cell proliferation, migration, and angiogenesis via miR-342/Wnt5a/β-catenin axis in vivo. H19 regulates glioma cell growth and metastasis through miR-342 to mediate Wnt5a/β-catenin signaling pathway, which provides new therapeutic targets for glioma treatment.

摘要

神经胶质瘤是最常见的恶性脑肿瘤,长链非编码 RNA(lncRNA)已被报道在神经胶质瘤的生长和血管生成中发挥重要作用。然而,lncRNA H19 在神经胶质瘤中的潜在机制尚不清楚。在本研究中,评估了 lncRNA H19 对神经胶质瘤细胞增殖、迁移和血管生成的影响。通过 RT-qPCR 或 Western blot 检测神经胶质瘤组织和细胞中 H19、miR-342 和 Wnt5a 的表达水平。双荧光素酶报告基因实验证实了 H19、miR-342 和 Wnt5a 之间的相互作用。通过集落形成、Transwell 和管形成实验分别分析细胞增殖、迁移和血管生成。免疫组织化学(IHC)用于检测血管生成相关因子 CD31。H19 和 Wnt5a 的表达在神经胶质瘤组织和细胞中显著上调,而 miR-342 的表达下调。此外,功能分析证实,敲低 H19 或过表达 miR-342 可抑制神经胶质瘤细胞的体外增殖、迁移和血管生成。此外,发现 H19 可直接靶向 miR-342 以促进神经胶质瘤细胞中 Wnt5a 的表达并激活 β-连环蛋白通路。此外,抑制 miR-342 或过表达 Wnt5a 可逆转 sh-H19 对神经胶质瘤生长和转移的抑制作用。此外,我们在体内验证了 H19 通过 miR-342/Wnt5a/β-连环蛋白轴调节神经胶质瘤细胞的增殖、迁移和血管生成。H19 通过调节 miR-342 来调节 Wnt5a/β-连环蛋白信号通路,从而调节神经胶质瘤细胞的生长和转移,为神经胶质瘤的治疗提供了新的治疗靶点。

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