猪miR-149靶基因预测及生物信息学分析

摘要/Abstract

摘要： 通过对猪miR-149进行靶基因预测及相关生物信息学分析，探索其影响猪颗粒细胞凋亡和卵泡闭锁的作用机制。〖KG-*3〗首先利用实时荧光定量PCR（qRT-PCR）检测ssc-miR-149在氧化应激组和正常组表达量的变化，通过转染miR-149 mimic并利用MTT和qRT-PCR检测细胞活性和凋亡基因的表达情况，同时预测ssc-miR-149与细胞凋亡相关基因的靶向关系；然后利用JASPAR、PROMO、TargetScan、miRanda、RNAhybrid、DAVID等软件和NCBI、miRBase、Ensembl等数据库对ssc-miR-149进行转录因子结合位点预测、保守性分析、靶基因预测、GO富集分析和KEGG信号通路富集分析。结果表明，氧化应激组ssc-miR-149的表达量极显著下调；但过表达ssc-miR-149时，能极显著提高细胞活性，并抑制H₂O₂诱导的凋亡相关基因（Caspase-3、PUMA和FAS）的表达；ssc-miR-149启动子区域具有SP1、p53等多个转录因子结合位点，其靶基因显著富集于TNF信号通路、Rap1信号通路和NOD样受体信号通路等。由此推测，ssc-miR-149受到SP1等多种转录因子调控，它可能通过抑制TNF、Rap1和NOD信号通路中的靶基因PUMA、Caspase-9和FAF-1来调控猪颗粒细胞的凋亡，进而影响猪的卵泡发育。

关键词: ssc-miR-149, 靶基因, 生物信息学, 颗粒细胞凋亡, 卵泡闭锁

张家庆，武松霞，任巧玲，王献伟，陈俊峰，王璟，白献晓，邢宝松. 猪miR-149靶基因预测及生物信息学分析[J]. 河南农业科学, 2019, 48(03): 123-131.

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