成牙骨质细胞矿化过程中可变剪接事件分析

Bioinformatics Analysis of Alternative Splicing Events during Mouse Cementoblast Mineralization

目的:生物信息学分析成牙骨质细胞矿化过程中可变剪接事件以及差异可变剪接基因富集到的生物学功能和信号通路。方法:小鼠成牙骨质细胞OCCM30矿化诱导0、7和14 d。实时荧光定量聚合酶链反应(qRT-PCR)检测骨钙素(bone gamma-carboxyglutamate protein,Bglap)和成骨细胞特异性转录因子(Osterix,Osx)的相对表达量来验证矿化结果。生物信息学分析可变剪接事件类型。Gene Ontology(GO)和Kyoto Encyclopedia of Genes and Genomes(KEGG)分别分析差异可变剪接基因可能富集到的生物学功能和信号通路。结果:Bglap和Osx在成牙骨质细胞矿化过程中表达量升高。与0 d相比,在矿化第7天和第14天时分别有1702个和1682个可变剪接事件发生。差异可变剪接基因主要与蛋白质丝氨酸/苏氨酸激酶活性、肽基赖氨酸修饰、Wnt信号通路和血管内皮生长因子信号通路等有关。结论:成功矿化诱导成牙骨质细胞。通过生信分析确定可变剪接事件以及差异可变剪接基因可能在成牙骨质细胞矿化过程中发挥作用,有望为今后牙骨质再生和牙周再生提供新的研究思路。

Objective:To bioinformational analyze the alternative splicing events and biofunctions and significant signaling pathways of differential alternative splicing genes during mouse cementoblasts(OCCM30)mineralization.Methods:OCCM30 were cultured in mineralized medium for 0,7,and 14 days.The relative gene expression levels of bone gamma-carboxyglutamate protein(Bglap)and Osterix(Osx)were detected by quantitative real-time polymerase chain reaction(qRT-PCR).Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)analysis were performed to enrich potential biofunctions and significant signaling pathways.Results:Compared with those on day 0,1702 and 1682 alternative splicing events were detected on day 7 and 14,respectively.Differential alternative splicing genes were associated with protein serine/threonine kinase activity,peptidyl-lysine modification,Wnt signaling pathway,and VEGF(vascular endothelial growth factor)signaling pathway and so on.Conclusion:Cementoblasts were successfully induced by mineralized medium.Using bioinformatics analysis,alternative splicing events were identified and differential alternative splicing genes may exert functions on cementoblast mineralization,which may provide a novel insight for cementogenesis and periodontal regeneration in the future.