Department of Cardiovascular Medicine, The Second Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330006, P.R. China.
Department of Hematology, The Second Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330006, P.R. China.
Int J Mol Med. 2018 Jul;42(1):489-496. doi: 10.3892/ijmm.2018.3595. Epub 2018 Mar 29.
Angiogenesis is essential for various biological processes, including tumor blood supply delivery, cancer cell growth, invasion and metastasis. Plasmacytoma variant translocation 1 (PVT1) long noncoding RNA (lncRNA) has been previously reported to affect angiogenesis of glioma microvascular endothelial cells by regulating microRNA (miR)‑186 expression level. However, the specific underlying molecular mechanism of PVT1 regulation of angiogenesis in vascular endothelial cells remains to be elucidated. The present study investigated the role of PVT1 in cell proliferation, migration and vascular tube formation of human umbilical vein endothelial cells (HUVECs) using MTT assay, Transwell migration assay and in vitro vascular tube formation assay, respectively. In order to determine the effect of miR‑26b on cell proliferation, migration and vascular tube formation of HUVECs, miR‑26 mimic or miR‑26b inhibitor were transfected into HUVECs. Reverse transcription‑quantitative polymerase chain reaction and western blotting were conducted to quantify the mRNA and protein expression levels of target genes. The present study confirmed that miR‑26b bound 3'‑untranslated region (3'‑UTR) and subsequently influenced gene expression level using dual luciferase reporter assay. The current study observed that PVT1 affected cell proliferation, migration and in vitro vascular tube formation of HUVECs. In addition, it was determined that PVT1 was able to bind and degrade miR‑26b to promote connective tissue growth factor (CTGF) and angiopoietin 2 (ANGPT2) expression. miR‑26b was also identified to have a suppressive role in cell proliferation, migration and in vitro vascular tube formation of HUVECs via binding 3'‑UTR regions and downregulating CTGF and ANGPT2 expression levels. The current findings may improve the understanding of the underlying mechanism of PVT1 contributing to angiogenesis of vascular endothelial cells and offer rationale for targeting PVT1 to treat angiogenesis dysfunction‑associated diseases, including cancer metastasis.
血管生成对于各种生物学过程至关重要，包括肿瘤血液供应的输送、癌细胞的生长、侵袭和转移。浆细胞瘤变异易位 1（PVT1）长链非编码 RNA（lncRNA）先前已被报道通过调节 microRNA（miR）-186 的表达水平来影响神经胶质瘤微血管内皮细胞的血管生成。然而，PVT1 调节血管内皮细胞血管生成的具体潜在分子机制仍有待阐明。本研究通过 MTT 测定法、Transwell 迁移测定法和体外血管形成测定法分别研究了 PVT1 在人脐静脉内皮细胞（HUVEC）增殖、迁移和血管形成中的作用。为了确定 miR-26b 对 HUVEC 增殖、迁移和血管形成的影响，将 miR-26 模拟物或 miR-26b 抑制剂转染到 HUVEC 中。逆转录-定量聚合酶链反应和蛋白质印迹法用于量化靶基因的 mRNA 和蛋白表达水平。本研究通过双荧光素酶报告基因测定证实了 miR-26b 结合 3' 非翻译区（3'UTR）并随后影响基因表达水平。本研究观察到 PVT1 影响 HUVEC 的增殖、迁移和体外血管形成。此外，研究发现 PVT1 能够结合并降解 miR-26b 以促进结缔组织生长因子（CTGF）和血管生成素 2（ANGPT2）的表达。miR-26b 还通过结合 3'UTR 区域并下调 CTGF 和 ANGPT2 表达水平，对 HUVEC 的增殖、迁移和体外血管形成具有抑制作用。本研究结果可能提高对 PVT1 促进血管内皮细胞血管生成的潜在机制的理解，并为针对 PVT1 治疗与血管生成功能障碍相关的疾病（包括癌症转移）提供理论依据。
