复制性衰老中全基因组染色质开放程度的变化-医学遗传学论文-基础医学论文-医学论文.docxVIP

复制性衰老中全基因组染色质开放程度的变化-医学遗传学论文-基础医学论文-医学论文 ——文章均为WORD文档，下载后可直接编辑使用亦可打印—— 　　摘 要：　为研究复制性衰老中表观遗传学的变化,以年轻代龄(PD26)和年老代龄(PD55)的人胚肺二倍体成纤维细胞(2BS)为材料,进行ATAC-seq测序,并使用Bowtie2、FastQC、MACS、deepTools、phastCons、R语言、HOMER以及基因本体论(Gene Ontology, GO)对测序结果进行深入分析和挖掘。结果显示, 2BS细胞染色质开放区域的保守性较强,其主要集中于启动子(promoter)区至转录起始位点(transcription start site, TSS),且年轻细胞与年老细胞的启动子区至转录起始位点的开放程度不同,细胞随着衰老在启动子区至转录起始位点的开放程度逐渐下降。进一步对年轻和年老细胞之间存在显着差异的靶基因进行GO分析发现,这些差异基因主要与细胞进程、代谢、生物性调节、结合功能、催化功能相关。本次研究发现衰老过程中染色质开放区减少、转录调控水平下降,提示复制性衰老与转录调控关系密切。 　　关键词：　复制性衰老; ATAC-seq测序; 转录调控; 染色质开放程度; 　　Abstract：　To study the epigenetic changes in replicative senescence, the ATAC-seq was performed in young(PD26) and senescent(PD55) 2BS cells. After sequencing, the results were analyzed by Bowtie2, FastQC,MACS, deepTools, phastCons, R language, HOMER and Gene Ontology(GO). The enrichment sites of open chromatin regions were highly conserved. Although the open regions were mainly enriched between the pro-moter and transcription start site(TSS), the enrichments in the young and the old are not the same, varying with aging of 2BS cells. Furthermore, GO analysis was performed to analyze the differentially expressed genes in different ages of 2BS cells. The genes were mainly clustered in cellular process, metabolic process,biological regulation, binding and so on. These results showed that the aging process was accompanied by declined degree of chromatin openness and decreased level of transcriptional regulation, which revealed the close relationship between the replicative senescence and transcriptional regulation. 　　Keyword：　replicative senescence; ATAC-seq; transcriptional regulation; opening of chromatin; 　　复制性衰老又称细胞衰老,当细胞周期停滞、凋亡抵抗以及部分基因如P16INK4a表达改变时,细胞出现衰老表型[1,2]。复制性衰老是整体衰老的基础,近期研究发现其具有拮抗癌症、使癌细胞得以清除的能力[3,4]。近年来,随着人口老龄化的发展及癌症等老年病发病率的增加,细胞衰老作为老年疾病的主要病理基础,已成为分子生物学研究的热点之一[5,6,7,8]。 　　在真核生物中,DNA围绕组蛋白分级折叠,形成核小体,而核小体则通过进一步紧密折叠形成高级结构染色质[9]。此时,根据细胞所处的状态,其基因组可划分为活跃表达的基因与相对沉默的基因。在DNA分级折叠的这一过程中,非活跃基因表达相关的结构区域相对封闭、核小体结构致密,呈转录抑制