Department of Oncology, Xiangya Hospital, Central South University, Changsha, China.
Department of Pathology, Xiangya Hospital, Central South University, Changsha, China.
Neoplasma. 2020 Jan;67(1):111-118. doi: 10.4149/neo_2019_190121N61. Epub 2019 Nov 26.
Glioma is one of the most common and aggressive malignant primary brain tumors with high recurrence rate and mortality rate and heavily depends on the angiogenesis. LncRNA H19 has many diverse biological functions, including the regulation of cell proliferation, differentiation and metabolism. Here, we aimed to investigate the molecular mechanism of lncRNA H19 affecting angiogenesis in glioma, which could help to uncover potential target for glioma therapy. RT-qPCR analysis was performed to detect the expression of lncRNA H19 and miR-138 in HEB, U87, A172 and U373 cell lines. MTT assay was used to evaluate the cell viability. To evaluate the migration and invasion after lncRNA H19 knockdown, Transwell and wound healing assay were employed. After lncRNA H19 knockdown, protein expression of HIF 1α and VEGF was detected by western blot and tube formation was assessed. For the prediction and validation of the interaction between lncRNA H19 and miR-138, bioinformatics and luciferase assay were performed. We investigated the regulatory roles and downstream molecular mechanisms of lncRNA H19 in glioma by knockdown H19, which inhibited the proliferation, migration and angiogenesis of glioma cells. Moreover, miR-138 acted as a target of H19 as detected by luciferase reporter assays. Meanwhile, HIF-1α was also a target of miR-138 and miR-138 could also regulate the proliferation, migration and angiogenesis of glioma cells by targeting HIF-1α and affecting the expression of VEGF in turn. Our findings identified an upregulated lncRNA H19 in glioma cells, which could promote proliferation, migration, invasion and angiogenesis via miR-138/HIF-1α axis as a ceRNA. This study provided a new opportunity to advance our understanding in the potential mechanism of lncRNA in glioma angiogenesis.
神经胶质瘤是最常见和侵袭性最强的恶性原发性脑肿瘤之一，具有高复发率和死亡率，严重依赖血管生成。LncRNA H19 具有许多不同的生物学功能，包括调节细胞增殖、分化和代谢。在这里，我们旨在研究 lncRNA H19 影响神经胶质瘤血管生成的分子机制，这有助于揭示神经胶质瘤治疗的潜在靶点。采用 RT-qPCR 分析检测 HEB、U87、A172 和 U373 细胞系中 lncRNA H19 和 miR-138 的表达。采用 MTT 分析评估细胞活力。采用 Transwell 和划痕愈合实验评估 lncRNA H19 敲低后细胞的迁移和侵袭能力。采用 Western blot 检测 lncRNA H19 敲低后 HIF 1α 和 VEGF 的蛋白表达，采用管形成实验评估管形成能力。通过生物信息学和荧光素酶报告基因实验进行 lncRNA H19 与 miR-138 相互作用的预测和验证。通过 lncRNA H19 敲低研究其在神经胶质瘤中的调控作用及下游分子机制，发现抑制 H19 可抑制神经胶质瘤细胞的增殖、迁移和血管生成。此外，荧光素酶报告基因实验检测到 miR-138 作为 H19 的靶点。同时，HIF-1α 也是 miR-138 的靶点，miR-138 还可以通过靶向 HIF-1α 并进而影响 VEGF 的表达来调节神经胶质瘤细胞的增殖、迁移和血管生成。本研究鉴定了神经胶质瘤细胞中上调的 lncRNA H19，其可通过 miR-138/HIF-1α 轴作为 ceRNA 促进增殖、迁移、侵袭和血管生成。本研究为深入了解 lncRNA 在神经胶质瘤血管生成中的潜在机制提供了新的机会。
