Department of Ophthalmology, Putuo Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai, 200062, China.
Department of Ophthalmology, Putuo Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai, 200062, China.
Exp Eye Res. 2019 Jul;184:38-47. doi: 10.1016/j.exer.2019.04.005. Epub 2019 Apr 16.
Conjunctivochalasis (CCH) is a common ocular disease, especially in aged people. However, the molecular mechanism of CCH on transcriptional level has been unclear. In this study, we characterized the transcriptional landscape of human conjunctiva and compared the transcriptome between normal persons (n = 10) and CCH patients (n = 11). Illumina RNA sequencing (RNA-seq) was performed to obtain transcriptional data, and these data were analyzed using various bioinformatics methods, including read mapping, the analysis of gene expression, gene ontology (GO) analysis and Kyoto Encyclopedia of Gene and Genomes (KEGG) metabolic pathway analysis. Additionally, expression patterns of 20 dysregupated genes were validated by qRT-PCR. RNA-seq result showed that clean ratios of 21 samples were more than 95% and more than 92% of all clean reads (32-41 million reads) were mapped to human genome sequence. There were 175 up-regulated genes and 582 down-regulated genes identified in CCH compared to normal persons. Among down-regulated genes in CCH, many genes were related with cell cycle and proliferation, such as BUB1, CCNB1, CCNB2 and CENPA, which might disturb cell growth and proliferation. In addition, several down-regulated genes were associated with keratinization and differentiation of epidermal cells, such as SPRR1A, SPRR1B, and CALML5. In over-expressed genes, CALML6 might play important roles on the development of CCH. The results of qRT-PCR confirmed the accuracy and credibility of RNA-Seq analysis. This study provided a lot of valuable information about pathogenic mechanism of CCH, which could be used to better study CCH in the future.
结膜松弛症（CCH）是一种常见的眼部疾病，尤其在老年人中较为常见。然而，CCH 在转录水平上的分子机制尚不清楚。在本研究中，我们对人结膜的转录组进行了特征描述，并比较了正常人和 CCH 患者（n=10）之间的转录组。我们使用 Illumina RNA 测序（RNA-seq）获得转录组数据，并使用各种生物信息学方法对这些数据进行分析，包括读映射、基因表达分析、基因本体（GO）分析和京都基因与基因组百科全书（KEGG）代谢途径分析。此外，通过 qRT-PCR 验证了 20 个差异表达基因的表达模式。RNA-seq 结果表明，21 个样本的清洁比均超过 95%，超过 92%的所有清洁读段（32-4100 万读段）均被映射到人类基因组序列上。与正常人相比，CCH 患者中有 175 个上调基因和 582 个下调基因。在 CCH 下调基因中，许多基因与细胞周期和增殖有关，如 BUB1、CCNB1、CCNB2 和 CENPA，这可能会干扰细胞生长和增殖。此外，一些下调基因与表皮细胞的角化和分化有关，如 SPRR1A、SPRR1B 和 CALML5。在过表达基因中，CALML6 可能在 CCH 的发生发展中发挥重要作用。qRT-PCR 的结果证实了 RNA-seq 分析的准确性和可信度。本研究为 CCH 的发病机制提供了大量有价值的信息，可用于今后更好地研究 CCH。
