| 刘华锋,王 超,张 伟,张 明,夏 毅.miR-613通过靶向VEGFA抑制神经胶质瘤细胞的增殖、侵袭和血管生成[J].,2021,(15):2823-2829 |
| miR-613通过靶向VEGFA抑制神经胶质瘤细胞的增殖、侵袭和血管生成 |
| MiR-613 Inhibits Proliferation, Invasion and Angiogenesis of Glioma Cells by Targeting VEGFA |
| 投稿时间：2020-12-27 修订日期：2021-01-23 |
| DOI：10.13241/j.cnki.pmb.2021.15.005 |
| 中文关键词: VEGFA 神经胶质瘤细胞 miR-613 |
| 英文关键词: VEGFA Glioma cells miR-613 |
| 基金项目:国家自然科学基金青年基金项目（81501115） |
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| 中文摘要: |
| 摘要 目的：旨在探究miR-613在胶质瘤中的表达及对细胞增殖、侵袭和血管生成的影响。方法：根据细胞转染将实验分组为对照miRNA组（Control组）、miR-613模拟物组（mimics组）和miR-613 mimics+VEGFA组（VEGFA组）。采用逆转录定量聚合酶链反应（RT-qPCR）检测胶质瘤细胞和组织中miR-613和VEGFA mRNA的表达水平；采用荧光素酶报告基因分析miR-613与血管内皮生长因子（VEGF）的关系；采用Western blotting检测VEGFA蛋白的表达水平；通过体外实验检测转染细胞的增殖能力、侵袭能力和管状形成能力。结果：与正常组织样本相比，胶质瘤I-II期组样本的肿瘤细胞呈现异形，具有深核染色，并且肿瘤细胞密度适度较低，而胶质瘤III-IV期组样本的肿瘤细胞的核分裂活跃，具有明显的微血管增殖和明显的细胞异型性；miR-613在胶质瘤I-IV期组织样本中显著降低（P<0.05）。在U87和U251细胞系的VEGFA-WT组中，与Control组相比，mimics组的荧光素酶活性显著降低（P<0.05）。与Control组相比，U87和U251细胞系中mimics组VEGFA的mRNA和蛋白表达水平均显著降低（P<0.05）。克隆形成实验、血管生成实验和细胞侵袭实验结果表明，与Control组相比，mimics组的克隆形成数量、细胞侵袭数、内皮细胞HUVEC的管状形成数和Ang-2蛋白表达水平均显著降低（P<0.05）；与mimics组相比，VEGFA组克隆形成数量、细胞侵袭数、内皮细胞HUVEC的管状形成数和Ang-2蛋白表达水平均显著升高（P<0.05）。结论：miR-613通过靶向VEGFA抑制了神经胶质瘤细胞的侵袭、增殖和血管生成，提示miR-613可能成为未来治疗胶质瘤的潜在靶点。 |
| 英文摘要: |
| ABSTRACT Objective: The aim of this study was to investigate the expression of mir-613 in glioma and its effect on cell proliferation, invasion and angiogenesis. Methods: According to cell transfection, the experiment was divided into control miRNA group (Control), miR-613 mimics group (miR-613) and miR-613 mimics + VEGFA group (miR-613+VEGFA). The expression of miR-613 in glioma cells and tissues was detected by reverse transcription quantitative polymerase chain reaction (RT-qPCR), and the relationship between miR-613 and vascular endothelial growth factor(VEGF) was analyzed by luciferase reporter gene. Then, the expression of VEGFA protein was detected by Western blotting, and the proliferation, invasion and tubular formation of transfected cells were detected in vitro. Results: Compared with normal tissue samples, the tumor cells of glioma stage I-II group showed heteromorphism, deep nuclear staining and moderately low density of tumor cells, while the tumor cells of glioma stage III-IV group had active mitosis, obvious microvascular proliferation and obvious cell atypia; miR-613 was significantly reduced in glioma stage I-IV tissue samples (P<0.05). In the VEGFA-WT group of U87 and U251 cell lines, compared with the Control group, the luciferase activity of the mimics group was significantly reduced (P<0.05). Compared with the Control group, the mRNA and protein expression levels of VEGFA in the mimics group in U87 and U251 cell lines were significantly lower (P<0.05). The results of clone formation experiment, angiogenesis experiment and cell invasion experiment showed that compared with the Control group, the number of clone formation, the number of cell invasion, the number of tube formation of endothelial cells HUVEC and the expression level of Ang-2 protein in the mimics group were significantly reduced (P<0.05); Compared with the mimics group, the number of clone formation, the number of cell invasion, the number of tube formation of endothelial cells HUVEC and the expression of Ang-2 protein in the VEGFA group were significantly increased (P<0.05). Conclusion: miR-613 inhibits the invasion, proliferation and angiogenesis of glioma cells by targeting VEGFA, suggesting that miR-613 may become a potential target for the treatment of glioma in the future. |
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