南方医科大学学报

南方医科大学学报 ›› 2024, Vol. 44 ›› Issue (3): 578-584.doi: 10.12122/j.issn.1673-4254.2024.03.20

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miR-26b-3p 靶向 CREB1 调控神经胶质瘤细胞的增殖、迁移及侵袭

黄秋虎，周 建，王子珍，杨 堃，陈政纲   

  1. 海南医学院第一附属医院神经外科，海南 海口 570102
  • 出版日期:2024-03-20 发布日期:2024-04-03

MiR-26-3p regulates proliferation, migration, invasion and apoptosis of glioma cells by targeting CREB1

HUANG Qiuhu, ZHOU Jian, WANG Zizhen, YANG Kun, CHEN Zhenggang   

  1. Department of Neurosurgery, First Affiliated Hospital of Hainan Medical University, Haikou 570102, China
  • Online:2024-03-20 Published:2024-04-03

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摘要： 目的 探讨miR-26b-3p靶向调控环磷酸腺苷效应元件结合蛋白1（CREB1）表达水平影响胶质瘤细胞增殖、迁移和侵袭能力的分子机制。方法 运用RT-qPCR和Western blotting检测不同级别胶质瘤中miR-26b-3p和CREB1的表达情况；生物信息学方法分析miR-26b-3p与CREB1结合的靶向序列。采用双荧光素酶报告基因检测miR-26b-3p对CREB1的靶向调控机制；将胶质瘤U251细胞分为对照组、miR-26b-3p mimic组及miR-26b-3p inhibitor组，采用Western blotting检测CREB1的表达变化，采用CCK-8法检测各组细胞增殖能力的影响，采用划痕实验检测各组细胞迁移能力的影响，采用Transwell检测各组细胞侵袭能力的影响，采用流式细胞术检测各组细胞凋亡的影响。结果 miR-26b-3p的表达随着胶质瘤级别的增加而降低（P<0.05），而CREB1的表达则逐渐增加，差异有统计学意义（P<0.05）；双荧光素酶报告基因结果显示miR-26b-3p可显著影响CREB1 3′UTR表达载体的荧光素酶活性，CREB1是miR-26b-3p下游靶基因。抑制miR-26b-3p表达可上调CERB1的表达，进而抑制细胞凋亡，促进胶质瘤细胞的增殖和侵袭。过表达miR-26b-3p可下调CERB1的表达，促进细胞凋亡，抑制胶质瘤细胞的增殖和侵袭（P<0.05）。结论 miR-26b-3p可靶向调控CREB1的表达调节胶质瘤细胞的凋亡、增殖、迁移和侵袭，进而参与胶质瘤的发生发展。

关键词: CREB1；miR-26b-3p；胶质瘤；增殖；迁移；侵袭

Abstract: Objective To investigate the regulatory role of miR-26b-3p in proliferation, migration and invasion of glioma. Methods The expressions of miR-26b-3p and cAMP-responsive element binding protein 1 (CREB1) in gliomas of different pathological grades were detected with RT-qPCR and Western blotting. Bioinformatic methods were used to analyze the target sequence of miRNA-26b-3p binding to CREB1, and dual luciferase gene reporter experiment was performed to explore the mechanism for targeted regulation of CREB1 by miR-26b-3p. Glioma U251 cells were treated with miR-26b-3p mimic or inhibitor, and the changes in CREB1 expression and cell proliferation, migration, invasion and apoptosis were determined with Western blotting, CCK-8 assay, wound healing assay, Transwell assay, and flow cytometry. Results The expression of miR-26b-3p decreased while CREB1 expression increased significantly as the pathological grade of gliomas increased (P<0.05). Dual luciferase gene reporter experiment confirmed that CREB1 was a downstream target of miR-26b-3p. Inhibition of miR-26b-3p significantly upregulated the expression of CERB1, suppressed apoptosis and promoted proliferation and invasion of glioma cells, and overexpression of miR-26b-3p produced the opposite effects (P<0.05). Conclusion MiR-26b-3p regulates CREB1 expression to modulate apoptosis, proliferation, migration and invasion of glioma cells, thereby participating in tumorigenesis and progression of glioma.

Key words: cAMP-responsive element binding protein 1; miR-26b-3p; glioma; proliferation; invasion; migration