Down-regulation and CpG island hypermethylation of CRYAA in age-related nuclear cataract
Down-regulation and CpG island hypermethylation of CRYAA in age-related nuclear cataract
复制标题
年龄相关性核性白内障中 CRYAA 的下调和 CpG 岛高甲基化
DOI：
10.1096/fj.12-213702
复制
发表时间：
2012-12-01
期刊：
影响因子：
4.8
通讯作者：
Lu, Yi
中科院分区：
文献类型：
--
作者：
Zhou, Peng;Luo, Yi;Lu, Yi
年龄相关性白内障(ARC)是老年人致盲的主要原因，αA-晶状体蛋白(CRYAA)的伴侣样活性被认为是维持眼晶状体透明度的关键。为了评估与ARC发病相关的CRYAA基因的表观遗传调控的潜在贡献，我们评估了DNA甲基化，这是一种影响基因表达的组织特异性遗传调节。逆转录-聚合酶链式反应和Western印迹分析CRYAA基因的表达。对15只老年性核性白内障患者和15只对照眼的CpG岛进行亚硫酸氢盐基因组测序，分析其甲基化状态。用去甲基化试剂ZeBularine研究CpG岛高甲基化与CRYAA表达下调的关系。年龄相关性核性白内障患者晶状体上皮细胞中CRYAA的mRNA和蛋白水平显著低于年龄匹配的对照组，这与CRYAA启动子的CpG岛高甲基化有关。使用DNA去甲基化药物治疗与恢复CRYAA的表达有关。比较DNA甲基化、mRNA和蛋白质水平，发现年龄相关性核性白内障与对照晶状体之间存在显着差异。这些证据表明，CRYAA在老年性核性白内障的晶状体上皮细胞中经历了表观遗传抑制。FASE B J.26,4897-4902(2012)。Www.fasebj.org
Age-related cataract (ARC) is the leading cause of blindness among older adults, and the chaperone-like activity of alpha A-crystallin (CRYAA) is considered to be critical for the maintenance of eye lens transparency. To assess the potential contribution of epigenetic regulation of CRYAA genes relevant to ARC pathogenesis, we evaluated DNA methylation, a tissue-specific genetic modulation that affects gene expression. Reverse-transcription PCR and Western blot were used to analyze the expression of CRYAA. Methylation status was analyzed by bisulfite genomic sequencing of the CpG islands in 15 eyes with age-related nuclear cataracts and 15 control eyes. The demethylating agent zebularine was used to investigate the relationship between hypermethylation of the CpG islands and down-expression of CRYAA. The mRNA and protein levels of CRYAA were significantly reduced in the lens epithelia of age-related nuclear cataract cases vs. age-matched controls, which corresponded to hypermethylation of the CpG island of CRYAA promoter. Treatment with a DNA-demethylating agent was associated with restoration of CRYAA expression. Comparing DNA methylation and mRNA and protein levels revealed significant differences between age-related nuclear cataract and control lenses. The evidence presented suggests that CRYAA undergoes epigenetic repression in the lens epithelia in age-related nuclear cataract.-Zhou, P., Luo, Y., Liu, X., Fan, L., Lu, Y. Down-regulation and CpG island hypermethylation of CRYAA in age-related nuclear cataract. FASEB J. 26, 4897-4902 (2012). www.fasebj.org