Research on the regulatory relationship and mechanism of miR-216b and CYB561D2 in gliomas based on bioinformatics analysis

doi:10.19854/j.cnki.1008-2425.2025.03.0008

Abstract

Abstract: Objective To investigate the regulatory relationship and mechanism between microRNA-216b (miR-216b) and its potential target gene Cytochrome b561 domain-containing protein 2 (CYB561D2) in gliomas.Methods Fifty-six pairs of glioma tissues and adjacent normal tissues were collected from Anqing Municipal Hospital.qRT-PCR and immunohistochemistry were used to detect the protein and mRNA expression of CYB561D2,and its correlation with miR-216b was analyzed.Bioinformatics databases were employed to predict miR-216b target genes,and dual-luciferase reporter assays were performed to validate the targeting relationship.Cell transfection,Western blotting,and qRT-PCR were used to investigate the regulatory effect of miR-216b on CYB561D2 expression in U251 and U87 glioma cells.Results Bioinformatics analysis showed that the 3'-UTR region of CYB561D2 contained conserved binding sites for miR-216b.Clinical tissue detection revealed that CYB561D2 mRNA (2.38±0.65 vs.1.00±0.21,P＜0.01) and protein (high expression rate:67.9% vs.14.3%,P＜0.01) levels in glioma tissues were significantly higher than those in adjacent normal tissues,and negatively correlated with miR-216b expression (r=-0.684,P＜0.01).Cell experiments confirmed that CYB561D2 protein was significantly upregulated in U251 and U87 cells compared with normal astrocytes.Transfection with miR-216b mimics significantly reduced CYB561D2 mRNA and protein expression in U251 and U87 cells (P＜0.01).Dual-luciferase reporter assays showed that miR-216b directly bound to the 3'-UTR region of CYB561D2 to inhibit its expression.Conclusion miR-216b directly targets CYB561D2 and inhibits its expression in gliomas,with a negative correlation between the two.The miR-216b/CYB561D2 axis may serve as a new direction for molecular diagnosis and targeted therapy of gliomas,although the in vivo functions and downstream mechanisms of this axis require further investigation.

Key words: Glioma, miR-216b, CYB561D2, Targeted regulation

摘要： 目的探究微小RNA-216b(miR-216b)及其潜在靶基因细胞色素b561结构域蛋白2(CYB561D2)在脑胶质瘤中的调控关系与作用机制。方法收集56例安庆市立医院脑胶质瘤组织及癌旁正常组织,利用qRT-PCR、免疫组织化学检测CYB561D2蛋白及mRNA表达,并分析其与miR-216b的相关性;采用生物信息学数据库预测miR-216b靶基因,双荧光素酶报告验证靶向关系;通过细胞转染、蛋白质免疫印迹和qRT-PCR,研究miR-216b对U251和U87胶质瘤细胞中CYB561D2表达的调控作用。结果生物信息学显示,CYB561D2 的3'-UTR区存在miR-216b保守结合位点。临床组织检测,胶质瘤组织中CYB561D2 蛋白(2.38±0.65 vs 1.00±0.21,P＜0.01),显著高于癌旁组织,且与miR-216b表达呈负相关(r=-0.684,P＜0.01)。细胞实验证实,与正常星形胶质细胞相比,U251和U87细胞中CYB561D2蛋白显著上调;转染miR-216b模拟物后,U251和U87细胞中CYB561D2的mRNA和蛋白表达均显著降低(P＜0.01)。双荧光素酶报告实验表明,miR-216b可直接结合CYB561D2的3'-UTR区抑制其表达。结论 miR-216b可直接靶向CYB561D2,抑制其在脑胶质瘤中的表达,二者呈负相关。miR-216b/CYB561D2轴可能成为脑胶质瘤分子诊断和靶向治疗的新方向,但该轴在体内的功能及下游机制需进一步研究。

关键词: 脑胶质瘤, CYB561D2, miR-216b, 靶向调控

CLC Number:

R739.41

Li Wentao, Zha Zhengjiang, Zhang Qingchao, Huang Chun, Wang Heng, Zhu Keshuai, Xu Shuai, Zhou Heping. Research on the regulatory relationship and mechanism of miR-216b and CYB561D2 in gliomas based on bioinformatics analysis[J]. Chinese Journal of Stereotactic and Functional Neurosurgery, 2025, 38(3): 178-182.

李文韬, 查正江, 张清超, 朱克帅, 徐帅, 周和平. 基于生物信息学分析miR-216b及CYB561D2在脑胶质瘤中的调控关系与作用机制研究[J]. 立体定向和功能性神经外科杂志, 2025, 38(3): 178-182.

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