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中国癌症防治杂志

中国癌症防治杂志 ›› 2020, Vol. 12 ›› Issue (5): 538-542.doi: 10.3969/j.issn.1674-5671.2020.05.10

• 基础研究 • 上一篇    下一篇

REST4 对胶质细胞瘤 U87细胞生物学行为的影响及其作用机制

  

  1. 天门市第一人民医院病理科；邯郸市第一医院肿瘤内科；天门市第一人民医院普外科
  • 出版日期:2020-10-25 发布日期:2020-10-25
  • 通讯作者: 牛东生 Ｅ-ｍａｉｌ：niuds3854@163.com
  • 基金资助:
    河北省医学科学研究重点课题计划项目（20181480）

Effect of REST4 on biological behavior of glioma U87 cells and its mechanism

  1. Department of Pathology, Tianmen First People's Hospital, ; Department of Oncology, the First Hospital of Handan
  • Online:2020-10-25 Published:2020-10-25
  • Contact: NIU Dongsheng E-mail: niuds3854@163.com

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摘要： 目的 探讨抑制元素l-沉默转录因子4 （repressor element 1-silencing transcription factor 4，REST4）基因对胶质细胞瘤U87细胞增殖、凋亡、迁移的影响及可能的作用机制。方法  将REST4-siRNA转染至胶质细胞瘤U87细胞（si-REST4组），同时设置siRNA阴性对照组（si-NC组）及空白对照组（Control组）。采用实时荧光定量PCR检测REST4 mRNA的表达水平，MTT法检测细胞增殖能力，Annexin V-FITC/PI 双染法检测细胞凋亡情况，划痕实验检测细胞迁移能力，Western blot检测U87细胞中REST4、PI3K、AKT、mTOR、p-PI3K、p-AKT和p-mTOR蛋白的表达水平。结果  转染后，与Control组及si-NC组相比，si-REST4组U87细胞中REST4 mRNA及蛋白表达水平下调（P<0.05），细胞增殖能力降低（P<0.05），细胞凋亡比例增加（P<0.05），细胞迁移率降低（P<0.05）。Western blot检测结果显示，与Control组及si-NC组相比，si-REST4组U87细胞PI3K、AKT、mTOR、p-PI3K、p-AKT、p-mTOR蛋白表达水平均降低（P<0.05）。结论 REST4可能通过 PI3K/AKT/mTOR信号通路抑制胶质细胞瘤U87细胞增殖、迁移并促进其凋亡。

关键词: 胶质细胞瘤, REST4, 增殖, 凋亡, 迁移, PI3K/AKT/mTOR信号通路

Abstract: Objective To investigate the effect of repressor element 1-silencing transcription factor 4 (REST4) gene on the proliferation, apoptosis and migration of glioma U87 cells and its possible mechanism. Methods REST4-siRNA was transfected into glioma U87 cells (si-REST4 group), while siRNA negative control group (si-NC group) and Control group were set up. The expression of REST4 was detected by the real-time quantitative PCR, the ability of cell proliferation was detected by MTT method, the cell apoptosis rate was detected by Annexin V-FITC/PI double staining, and the ability of cell migration was detected by scratch test. The expressions of REST4, PI3K, AKT, mTOR, p-PI3K, p-AKT and p-mTOR proteins in U87 cells were detected by Western blot. Results Compared with the Control group and si-NC group, the REST4 mRNA and protein levels of U87 cells in the si-REST4 group were down-regulated (P<0.05), the ability of cell proliferation decreased (P<0.05), the proportion of cell apoptosis increased(P<0.05), and the cell migration decreased (P<0.05). Western blot showed that compared with the Control group and the si-NC group, the protein expression levels of PI3K, AKT, mTOR, p-PI3K, p-AKT and p-mTOR of U87 cells in the si-REST4 group decreased (P<0.05). Conclusions REST4 may inhibit the proliferation and migration of glioma U87 cells and promote their apoptosis through PI3K/AKT/mTOR signal pathway.

Key words: Glioma, REST4, Proliferation, Apoptosis, Migration, PI3K/AKT/mTOR signaling pathway

中图分类号: 

  • R730.264