衰老是老年性疾病发生的主要原因之一，已有研究表明长链非编码RNA（lncRNAs）参与调控细胞衰老进程。课题组前期对复制性衰老和氧化应激早熟性衰老的人成纤维细胞（NHF）进行了lncRNA芯片分析，lncRNA CAR intergenic 10是我们从芯片中筛选出的在两种衰老方式中均低表达的lncRNA，初步功能实验显示lncRNA CAR 10可延缓NHF细胞衰老，但具体作用机制尚待深入探讨。本课题拟采用逆转录病毒表达载体及siRNA干扰技术确定lncRNA CAR 10在NHF细胞衰老中的功能；利用RNA-pulldown、RNA-RIP等技术结合蛋白质谱等手段筛选、鉴定lncRNA CAR 10直接结合的蛋白分子及可能调控的靶基因；阐明CAR 10调控细胞衰老的下游通路和分子机制。本研究从lncRNA角度阐述细胞衰老的分子机制,将为延缓并减少老年性疾病的发生提供重要依据。

Cellular senescence contributes to aging and/or age-related diseases. While p53/p21 and p16/pRB pathways are well known to play critical roles in cellular senescence, the molecular basis for its initiation and maintenance remains to be fully elucidated. Long non-coding RNAs (lncRNAs), which were recently found to be pervasively transcribed in the genome, are emerging as new players in gene expression regulation. LncRNAs can interact with its partner DNA, RNA, and/or protein to exert their function. However, knowledge regarding the contributions of lncRNAs to cellular senescence remains scarce.. In order to explore the role of the lncRNAs in cellular senescence, we screened for the differentially expressed lncRNAs among replicative senescent normal human fibroblasts (NHF), H2O2-induced premature senescent NHF and young NHF by microarray and validated the candidate lncRNAs by Q-PCR. LncRNA CAR intergenic 10 was found to be lowly expressed in senescent NHF cells. Silencing lncRNA CAR 10 resulted in an increased percentage of senescent NHF cells. Thus lncRNA CAR 10 possibly helps to prevent the onset of cellular senescence. To test this, we will assess the effect of lncRNA CAR 10 on cellular senescence by overexpressing or silencing the lncRNA in NHF; interrogate the partner proteins through RNA-pulldown and RNA-RIP. This study will show whether lncRNA CAR 10 delays cellular senescence and provide important clues about its molecular partners and its biological function.