Department of Neurosurgery, Hunan Provincial People's Hospital, No.61, Jiefang West Road, Changsha, 410005, Hunan, People's Republic of China.
Mol Cell Biochem. 2020 May;468(1-2):69-82. doi: 10.1007/s11010-020-03712-y. Epub 2020 Apr 1.
Glioma is characterized by high morbidity, high mortality and poor prognosis. Recent studies exhibited that lncRNA CCAT2 is overexpressed in glioma and promotes glioma progression, but the specific molecular biological mechanism remains to be determined. We performed qRT-PCR to evaluate the expression of related genes, Western blotting analysis to measure protein levels, colony formation assay to detect the proliferative ability of glioma cells, flow cytometry to measure cell apoptosis, bioinformatics analysis and dual luciferase assay to verify the binding sites and the targeted regulatory relationship in A172 and U251 cell lines and tube formation assay to determine endothelial angiogenesis. LncRNA CCAT2 and VEGFA were highly expressed, while miR-424 was expressed at low levels in NHA cells. Furthermore, knockdown of lncRNA CCAT2 decreased cell proliferation, increased cell apoptosis and inhibited endothelial angiogenesis in glioma. Moreover, lncRNA CCAT2 shared a complementary sequence with miR-424 which in turn directly bound to the 3'-UTR of VEGFA. Further investigation indicated that lncRNA CCAT2 promoted cell proliferation and endothelial angiogenesis by inducing the PI3K/AKT signalling pathway in glioma. The oncogenic lncRNA CCAT2 is highly associated with the development of glioma and exerts its function by upregulating VEGFA via miR-424.
神经胶质瘤的发病率、死亡率和预后均较差。最近的研究表明，lncRNA CCAT2 在神经胶质瘤中表达上调，促进神经胶质瘤的进展，但具体的分子生物学机制仍有待确定。我们通过 qRT-PCR 评估相关基因的表达，通过 Western blot 分析测量蛋白水平，通过集落形成实验检测神经胶质瘤细胞的增殖能力，通过流式细胞术测量细胞凋亡，通过生物信息学分析和双荧光素酶报告基因实验验证 A172 和 U251 细胞系中的结合位点和靶向调控关系，通过管形成实验确定血管内皮的生成。在 NHA 细胞中，lncRNA CCAT2 和 VEGFA 表达上调，而 miR-424 表达水平较低。此外，lncRNA CCAT2 的敲低降低了神经胶质瘤细胞的增殖，增加了细胞凋亡，并抑制了血管内皮的生成。此外，lncRNA CCAT2 与 miR-424 具有互补序列，而 miR-424 又直接结合到 VEGFA 的 3'UTR 上。进一步的研究表明，lncRNA CCAT2 通过诱导神经胶质瘤中的 PI3K/AKT 信号通路促进细胞增殖和血管内皮生成。致癌性 lncRNA CCAT2 与神经胶质瘤的发生发展密切相关，通过上调 miR-424 来发挥其功能。
