LncRNA-AC125421.1的异常表达在寻常型银屑病发病中的作用及其分子机制探讨

Psoriasis is a polygenic, common chronic inflammatory disease characterized by skin lesions. Due to its unclear pathogenesis, psoriasis still lacks effective treatment. Long non-coding RNA (lncRNA) plays a very important role in the development of human disease and attracts more and more concerns at home and abroad, but its role in psoriasis has not been clarified. In previous work, 10 lesional and non-lesional tissues of psoriasis vulgaris and 10 healthy skin tissues were collected and then sequenced by high-throughput RNA sequencing (RNA-seq). We verified the most differentially expressed lncRNAs using quantitative real-time PCR in skin samples from 50 patients with psoriasis vulgaris and found that lncRNA-AC125421.1 was overexpressed. Subcellular localization demonstrated that lncRNA-AC125421.1 was in the cytoplasm of in vivo cultured HaCaT keratinocytes. In the present study, through in situ hybridization techniques, RNA-Binding Protein Immunoprecipitation (RIP), luciferase reporter gene systems and bioinformatics analysis, we extensively explored the biological function and tried to reveal the underlying molecular mechanism of lncRNA-AC125421.1 in psoriasis. The findings of this study will help to better understand the molecular mechanisms of psoriasis and to lay a foundation for developing new therapeutic target.

银屑病是一种由多基因遗传决定的慢性炎症性皮肤病。由于发病机制不清，目前仍缺乏有效的治疗手段。探索长链非编码RNA（lncRNA）在人类疾病中的作用是目前国内外的研究热点。我们在前期的实验工作中，对10例寻常型银屑病患者的皮损和非皮损组织RNA样本进行了高通量测序，通过实时定量PCR法对其中最为显著差异表达的lncRNAs在50例寻常型银屑病患者的皮肤样本中进行了验证，发现lncRNA-AC125421.1呈异常高表达。在角质形成细胞系HaCaT中对lncRNA-AC125421.1进行亚细胞定位后，发现其存在于细胞质中。本项目在前期研究工作的基础上，应用原位杂交技术、RIP技术、荧光素酶报告基因系统和生物信息学分析等手段，深入探讨lncRNA-AC125421.1在银屑病发病中的作用并揭示其作用分子机制。本项目的研究成果将有助于更好地理解银屑病的发病机制，为开发银屑病新的治疗靶点打下基础。

银屑病是一种以角质形成细胞高度增殖与异常分化为特征的慢性炎症性皮肤病。长链非编码RNA（lncRNA）可以在DNA修饰、转录以及翻译等多个层面参与角质形成细胞的功能调节。本研究的目的是寻找银屑病中异常表达的lncRNA以及探索其潜在的作用机制。方法：选取了5例寻常型银屑病患者的皮损和年龄、性别匹配的整形外科手术切取剩余的正常皮肤组织5份，采用lncRNA和mRNA共表达芯片技术对其中KC转录组中差异表达的lncRNA和mRNA进行筛选和分子功能注释。利用Starbase生物信息学分析预测lncRNA所靶向的miRNA， 扩大样本量，在银屑病皮损、非皮损组织和正常皮肤组织中通过qRT-PCR对其中差异表达的lncRNA、miRNA和mRNA表达情况进行验证。结果：基因共表达芯片结果显示银屑病皮损与正常皮损有48个差异表达的lncRNA和17个差异表达的mRNA，其中 lncRNA AGXT2L1上调2.05倍，NAP1L1上调1.66倍。Starbase生物信息学分析预测显示，在所有上调表达的LncRNA中，仅有lncRNA-AGXT2L1特异性地单一指向靶向miR-484。继续扩大样本量，发现银屑病皮损较正常组织lncRNA AGXT2L1表达上升，miRNA-484表达下降，NAP1L1表达上升。结论：银屑病病皮损中lncRNA-AGXT2L1异常高表达，其可能通过与NAP1L1竞争性结合miRNA-484从而导致银屑病的角质形成细胞过度增殖。