TargetScan筛选退变椎间盘髓核细胞凋亡相关lncRNA的研究Target Scan as a tool in screening of apoptosis related long non-coding RNA in degenerative intervertebral nucleus pulposus cell
王毅峰;宋庆鑫;黄轩;陈智;于利洪;吴水祥;沈洪兴;
摘要(Abstract):
[目的]寻找退变椎间盘髓核细胞凋亡相关lncRNA,为髓核细胞凋亡的分子机制研究提供指导。[方法]比较miRNA靶基因预测方法,选择灵敏性和特异性均较高、包含不保守区域分析的Target Scan作为预测工具。基于现有lncRNA的序列信息,运行Target Scan的perl脚本,替换mRNA序列信息为lncRNA的序列信息,寻找miRNA的靶lncRNA。根据Target Scan预测结果,综合评估miRNA结合位点数量,靶lncRNA长度,Context+Score评分等指标,筛选目标lncRNA。[结果]文献回顾发现,hsa-miR-27a-3p是退变椎间盘髓核细胞凋亡的关键分子,退变髓核细胞中共有1 052个lncRNA存在差异表达(Fc值≥2,P<0.05),其中有186个lncRNA表达下调。检索Ensemble数据库、UCSC基因浏览器和miRbase数据库,明确差异表达的lncRNA的序列及定位信息,将序列定位明确的117个表达下调的lncRNA纳入研究,筛选退变髓核细胞差异表达lncRNA中含有hsa-miR-27a-3p结合位点的lncRNA。共有28个差异表达的lncRNA含有47个hsa-miR-27a-3p结合位点,其中,ENST00000486904和ENST00000408017的Context+Score百分位数≥90,ENST00000450202长度为459 bp,含有3个hsa-miR-27a-3p结合位点,ENST00000440936、ENST00000441356、ENST00000455216和ENST00000498840分别含有2个hsa-miR-27a-3p结合位点,可能通过吸附miR-27a在椎间盘退变过程中抑制PI3K介导的髓核细胞凋亡。[结论]应用Target Scan数据库进行生物信息学分析,可以有效预测退变椎间盘髓核细胞凋亡相关lncRNA,为进一步的实验研究提供良好的指导。
关键词(KeyWords): 长链非编码RNA;椎间盘退变;细胞凋亡;生物信息学分析
基金项目(Foundation): 上海市新百人计划项目(编号:XBR201399);; 长海医院“1255”学科建设计划项目(编号:CH125540200)
作者(Author): 王毅峰;宋庆鑫;黄轩;陈智;于利洪;吴水祥;沈洪兴;
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