铜死亡相关基因FDX1调控骨关节炎软骨细胞增殖的分子机制研究  

Molecular mechanism of cuproptosis-related gene FDX1 in regulating chondrocyte proliferation in osteoarthritis

作　　者：蔡灵敏 杨娅芹 郭翱[1] CAI Lingmin;YANG Yaqin;GUO Ao(Department of Sports Medicine,Taizhou Orthopedics Hospital,Wenling,Zhejiang 317500,China;Department of Orthopedics,Taizhou Orthopedics Hospital,Wenling,Zhejiang 317500,China)

机构地区：[1]浙江省台州骨伤医院运动医学科,温岭317500 [2]浙江省台州骨伤医院正骨科,温岭317500

出　　处：《浙江中西医结合杂志》2025年第2期112-120,共9页Zhejiang Journal of Integrated Traditional Chinese and Western Medicine

基　　金：浙江省温岭市社会发展科技项目(No.2023S00076)。

摘　　要：目的利用生物信息学构建铜死亡基因相关ceRNA调控网络并验证其对骨关节炎(OA)的影响。方法从GEO数据库GSE1919、GSE55235下载OA m RNA测序数据,利用R软件筛选数据集的差异表达基因(DEGs),与铜死亡相关基因(CRGs)取交集,获得与OA相关的DEG人铁氧还原蛋白1(FDX1)。从GEO数据库GSE105027下载OA miRNA测序数据,利用R软件筛选数据集的差异表达miRNA,与利用Starbase数据库预测FDX1可能存在调控关系的miRNA取交集,得到在OA样本中下调,且与FDX1互作的miRNA。从GEO数据库GSE175959下载OA circRNA测序数据,利用R软件筛选数据集的差异表达circRNA,与利用Starbase数据库预测mi R-514a-3p可能存在调控关系的circRNA取交集,得到在OA样本中上调,且与miRNA互作的circRNA。利用cytoscape软件构建ceRNA调控网络。Western blot实验检测组织中FDX1的表达水平。CCK-8检测细胞增殖、流式细胞术检测细胞凋亡。RT-qPCR检测circRNA_001549表达情况,RNA免疫沉淀分析circRNA_001549与mi R-514a-3p结合情况,双荧光素酶报告实验验证miR-514a-3p与FDX1结合情况。结果通过正常样本与OA样本间的差异分析,得到了473个差异基因,其中221个上调基因,252个下调基因。在与10个CRGs取交集后,得到1个铜死亡相关的差异基因FDX1。与正常组相比,OA组FDX1表达水平显著升高[(5.78±0.43)比(4.98±0.36),P<0.01]。通过ss GSEA算法发现OA组的B cells、Check-point、Cytolytic activity、HLA、Inflammation-promoting、Macrophages、Mast cells、Neutrophils、Parainflammation、T cell co-inhibition、T cell co-stimulation、T helper cells、TIL、TypeⅠIFN Reponse和TypeⅡIFN Reponse免疫细胞相对浸润丰度和功能评分显著升高,而aDCs、DCs和Treg显著降低。FDX1的表达水平与Macrophages和DCs免疫细胞浸润水平显著正相关。Starbase数据库预测的mi RNA与差异mi RNA取交集后,得到与FDX1互作的mi R-514a-3p,Starbase数据库预测的circRNA与差异circRNA取交集后,�Objective To construct a ceRNA regulatory network involving cuproptosis-related genes using bioinformatics and validate its effect on osteoarthritis(OA).Methods OA mRNA sequencing data were downloaded from GEO databases GSE1919 and GSE55235,and the differentially expressed genes(DEGs)in the dataset were selected using R software.Human ferredoxin 1(FDX1),a cuproptosis-related gene(CRG)associated with OA,was identified by intersecting DEGs with CRGs.OA miRNA sequencing data were downloaded from the GEO database GSE105027,and differentially expressed miRNAs were identified using R software.These miRNAs were intersected with miRNAs predicted by the Starbase database to have a potential regulatory relationship with FDX1,identifying downregulated miRNAs in OA samples that interact with FDX1.OA circRNA sequencing data were downloaded from the GEO database GSE175959,and R software was used to screen the dataset for differentially expressed circRNAs.circRNAs that were up-regulated in the OA samples and interacted with miR-514a-3p were obtained by intersecting the differentially expressed circRNAs with those predicted to have a possible regulatory relationship with miR-514a-3p using the Starbase database.A ceRNA regulatory network was constructed using Cytoscape software.The expression levels of FDX1 in tissues were detected using Western blot assay.Cell proliferation was measured using CCK-8 assay and cell apoptosis was assessed using flow cytometry.circRNA_001549 expression was detected using RT-qPCR,the binding of circRNA_001549 to miR-514a-3p was analyzed using RNA immunoprecipitation,and the binding between miR-514a-3p and FDX1 was validated using dual-luciferase reporter assay.Results By analyzing the difference between normal and OA samples,a total of 473 differential genes were identified,including 221 up-regulated genes and 252 down-regulated genes.After intersecting with 10 CRGs,FDX1,one cuproptosis-related differentially expressed gene was obtained.Compared with the normal group,the expression levels of FDX1 in

关 键 词：FDX1 骨关节炎 ceRNA网络 铜死亡相关基因 

分 类 号：R684.3[医药卫生—骨科学]