Szpakowski Sebastian, Sun Xueguang, Lage José M, Dyer Andrew, Rubinstein Jill, Kowalski Diane, Sasaki Clarence, Costa Jose, Lizardi Paul M
Interdepartmental Program in Computational, Biology and Bioinformatics, Yale University School of Medicine, Room LH-208, 310 Cedar Street, New Haven, CT 06520, USA.
Gene. 2009 Dec 15;448(2):151-67. doi: 10.1016/j.gene.2009.08.006. Epub 2009 Aug 21.
Close to 50% of the human genome harbors repetitive sequences originally derived from mobile DNA elements, and in normal cells, this sequence compartment is tightly regulated by epigenetic silencing mechanisms involving chromatin-mediated repression. In cancer cells, repetitive DNA elements suffer abnormal demethylation, with potential loss of silencing. We used a genome-wide microarray approach to measure DNA methylation changes in cancers of the head and neck and to compare these changes to alterations found in adjacent non-tumor tissues. We observed specific alterations at thousands of small clusters of CpG dinucleotides associated with DNA repeats. Among the 257,599 repetitive elements probed, 5% to 8% showed disease-related DNA methylation alterations. In dysplasia, a large number of local events of loss of methylation appear in apparently stochastic fashion. Loss of DNA methylation is most pronounced for certain members of the SVA, HERV, LINE-1P, AluY, and MaLR families. The methylation levels of retrotransposons are discretely stratified, with younger elements being highly methylated in healthy tissues, while in tumors, these young elements suffer the most dramatic loss of methylation. Wilcoxon test statistics reveals that a subset of primate LINE-1 elements is demethylated preferentially in tumors, as compared to non-tumoral adjacent tissue. Sequence analysis of these strongly demethylated elements reveals genomic loci harboring full length, as opposed to truncated elements, while possible enrichment for functional LINE-1 ORFs is weaker. Our analysis suggests that, in non-tumor adjacent tissues, there is generalized and highly variable disruption of epigenetic control across the repetitive DNA compartment, while in tumor cells, a specific subset of LINE-1 retrotransposons that arose during primate evolution suffers the most dramatic DNA methylation alterations.
近50%的人类基因组含有最初源自移动DNA元件的重复序列,在正常细胞中,这一序列区域受到涉及染色质介导的抑制作用的表观遗传沉默机制的严格调控。在癌细胞中,重复DNA元件会发生异常去甲基化,导致沉默作用可能丧失。我们采用全基因组微阵列方法来测量头颈部癌症中的DNA甲基化变化,并将这些变化与相邻非肿瘤组织中的变化进行比较。我们观察到与DNA重复序列相关的数千个CpG二核苷酸小簇存在特定变化。在检测的257,599个重复元件中,5%至8%显示出与疾病相关的DNA甲基化改变。在发育异常中,大量局部甲基化缺失事件以明显随机的方式出现。DNA甲基化缺失在SVA、HERV、LINE-1P、AluY和MaLR家族的某些成员中最为明显。逆转录转座子的甲基化水平呈离散分层,较年轻的元件在健康组织中高度甲基化,而在肿瘤中,这些年轻元件的甲基化缺失最为显著。Wilcoxon检验统计显示,与非肿瘤相邻组织相比,灵长类LINE-1元件的一个子集在肿瘤中优先发生去甲基化。对这些强烈去甲基化元件的序列分析揭示了含有全长而非截短元件的基因组位点,而功能性LINE-1开放阅读框的可能富集较弱。我们的分析表明,在非肿瘤相邻组织中,整个重复DNA区域的表观遗传控制普遍且高度可变地受到破坏,而在肿瘤细胞中,灵长类进化过程中出现