生物学杂志

生物学杂志 ›› 2025, Vol. 42 ›› Issue (5): 46-.doi: 10.3969/j.issn.2095-1736.2025.05.046

• 研究报告 • 上一篇    下一篇

miR-556影响人脑胶质母细胞瘤细胞生长及其转录组学分析

邢丽波1， 柴春娥2， 王尔鸿2， 王子言1， 陆东东1   

  1. 1. 同济大学 生命科学与技术学院， 上海 200092; 2. 同济大学 医学院， 上海 200331
  • 出版日期:2025-10-18 发布日期:2025-10-14
  • 作者简介:邢丽波，博士研究生，高级工程师，研究方向为分子生物学，E-mail：xinglibo@tongji.edu.cn
  • 基金资助:
    同济大学青年女教师成才资助金项目（08002150256）； 同济大学本科生创新创业能力拓展项目（X2022165）

Effect of miR-556 on the growth of human glioblastoma cells and its transcriptome analysis

XING Libo1, CHAI Chun’e2, WANG Erhong2, WANG Ziyan1, LU Dongdong1   

  1. 1. School of Life Sciences and Technology, Tongji University, Shanghai 200092, China;
    2. School of Medicine, Tongji University, Shanghai 200331, China
  • Online:2025-10-18 Published:2025-10-14

摘要： 研究旨在探究miR-556对人脑胶质母细胞瘤细胞（U-87 MG）生长、基因转录水平和基因功能的影响。通过慢病毒感染U-87 MG细胞，筛选稳定过表达miR-556的细胞系；通过定量和普通RT-PCR分别验证细胞中前体miR-556（Pre-miR-556）和成熟miR-556的表达量；通过CCK-8测定细胞的体外增殖能力；通过结晶紫染色测定细胞集落形成能力；通过高通量RNA测序（RNA sequencing， RNA-seq）技术发现细胞中基因组转录水平及信号通路的改变。成功构建稳定表达miR-556的U-87 MG细胞系；与对照组（rLV）相比，实验组（rLV-miR-556）U-87 MG细胞的体外增殖能力和集落形成能力显著增加（P<0.01）；与rLV组相比，rLV-mir-556组显示2028个差异基因，其中，165个基因上调表达，1863个基因下调表达；miR-556主要通过上调HMOX1、SERPINB2、ATOH8和下调MACF1、PEG10、HIF1A等基因的转录，从而改变基因表达（转录）、跨膜运输、RNA聚合酶Ⅱ转录、翻译后蛋白的修饰、蛋白质代谢、免疫系统、疾病等信号通路。miR-556提高了人脑胶质母细胞瘤细胞的体外生长能力，影响了人脑胶质母细胞瘤细胞中基因的转录水平和功能，研究为肿瘤的防治提供基础资料。

关键词: miR-556, 人脑胶质母细胞瘤, 慢病毒表达质粒, 细胞增殖, 转录组学

Abstract: This study investigated how miR-556 affects proliferation, genome-wide transcription and gene-function networks in human glioblastoma U-87 MG cells. A miR-556-overexpressing line was generated by lentiviral transduction and puromycin selection; precursor and mature miR-556 levels were quantified by qRT-PCR and standard RT-PCR. Cell proliferation and clonogenic capacity were assessed with CCK-8 and crystal-violet assays, respectively. RNA-seq was used to map transcriptional and pathway changes. Relative to the control (rLV) group, the miR-556-overexpressing (rLV-miR-556) group exhibited significantly increasedin vitroproliferation and colony formation (P<0.01). RNA-seq revealed 2028 differentially expressed genes (165 up- and 1863 down-regulated) between the two groups. miR-556 up-regulatedHMOX1,SERPINB2andATOH8while down-regulatingMACF1,PEG10andHIF1A, thereby modulating pathways related to gene transcription, transmembrane transport, RNA polymerase Ⅱ activity, post-translational modification, protein metabolism, immune responses and disease-related pathways. These findings indicate that miR-556 enhances glioblastoma cell growth and reshapes the transcriptional landscape, providing a basis for future intervention strategies.

Key words: MiR-556, human glioblastoma cells, lentivirus expression plasmid, cell proliferation, transcriptomics

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