基于基因表达谱分析成肌细胞应力敏感基因及其潜在功能  

Gene expression profiling analysis of stress-sensitive genes and their potential functions in myoblasts

作　　者：郑瑶 于少洋 阎潇[1,2] 李建平 张强[1,2] 袁晓[1,2] ZHENG Yao;YU Shao-yang;YAN Xiao;LI Jian-ping;ZHANG Qiang;YUAN Xiao(Department of Orthodontics,Affiliated Hospital of Qingdao University,Qingdao 266003;School of Stomatology,Qingdao University,Qingdao 266023,Shandong Province,China)

机构地区：[1]青岛大学附属医院口腔正畸科,山东青岛266003 [2]青岛大学口腔医学院,山东青岛266023

出　　处：《上海口腔医学》2025年第1期7-13,共7页Shanghai Journal of Stomatology

基　　金：国家自然科学基金(32171303);山东省医药卫生科技项目合同书(202308031256)。

摘　　要：目的:筛选成肌细胞内应力敏感基因,揭示功能矫形力诱导面颌部肌肉改建的潜在靶基因及其调控机制。方法:采用基因芯片技术分析成肌细胞内的差异表达基因(differentially expressed genes,DEGs)。通过基因本体论(Gene Ontology,GO)和京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)通路对DEGs进行富集分析。采用实时荧光定量PCR验证DEGs。采用蛋白免疫印迹、透射电镜(transmission electron microscopy,TEM)和激光扫描共聚焦显微镜(confocal laser scanning microscope,CLSM)检测应力对成肌细胞自噬的影响。采用SPSS 17.0软件包对数据进行统计学分析。结果:筛选成肌细胞内DEGs 1410个,其中,上调788个,下调622个。GO分析显示,DEGs主要富集于信号转导、生物聚合物代谢和蛋白质代谢等过程。KEGG结果显示,DEGs主要富集于细胞外基质受体相互作用通路、癌症通路、MAPK信号通路、黏着斑激酶和溶酶体等信号通路。TEM和CLSM均观察到应力促进自噬小体的形成。蛋白免疫印迹显示,应力可促进自噬相关分子Beclin-1和LC-3II的表达。以上过程均可被雷帕霉素促进或被3-MA抑制。结论:自噬可能在周期性张应力诱导成肌细胞命运调控过程中起重要作用。PURPOSE:To screen the stress-sensitive genes in myoblasts and reveal the potential target genes and their regulatory mechanisms of facial muscle remodeling induced by functional orthopaedic force.METHODS:The procedure involved the use of gene microarray technology to identify the differentially expressed genes(DEGs)in myoblasts.DEGs were then categorized by Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses.Furthermore,real-time quantitative PCR(qRT-PCR)was used to verify the DEGs.Western blot,transmission electron microscopy(TEM),and confocal laser scanning microscope(CLSM)were employed to detect the effect of stress on autophagy in myoblasts.The data were analyzed by SPSS 17.0 software package.RESULTS:A total of 1410 DEGs were identified in stretched myoblasts,with 788 up-regulated and 622 down-regulated genes.GO enrichment analysis indicated that DEGs were primarily involved in signal transduction,biopolymer metabolic process,and protein metabolic process.KEGG analysis revealed that DEGs were primarily associated with ECM-receptor interaction,pathway in cancer,MAPK signaling pathway,focal adhesion and lysosome.Both TEM and CLSM showed that stress could promote the formation of autophagosomes,and Western blot demonstrated that stress could promote the expression of autophagy-related molecules Beclin-1 and LC3-II.Rapamycin could enhance all the above processes,while 3-MA could inhibit them.CONCLUSIONS:Autophagy may play an important role in the regulation of myoblast fate induced by cyclic tensile stress.

关 键 词：周期性张应力 成肌细胞 基因芯片 信号通路 自噬 

分 类 号：R783[医药卫生—口腔医学]