Department of Dermatology, Qilu Hospital of Shandong University, Jinan 250012, Shandong, PR China.
Department of Dermatology, Qilu Hospital of Shandong University, Jinan 250012, Shandong, PR China.
Exp Cell Res. 2018 Feb 15;363(2):243-254. doi: 10.1016/j.yexcr.2018.01.014. Epub 2018 Jan 13.
Competitive endogenous RNAs (ceRNAs) regulate RNA transcripts by competing for shared miRNAs and play critical roles in disease development. Psoriasis is a long-lasting, recurring chronic inflammatory skin disease characterized by hyperproliferation of keratinocytes. The keratinocyte response is triggered by the activation of inflammatory cytokines, like interleukin-22 (IL-22). We used lncRNA array analysis to detect differentially expressed lncRNAs in skin (HaCaT) cells treated with or without IL-22. We used hematoxylin and eosin (H&E) staining to determine the pathological changes in skin cells and immunohistochemistry to evaluate the expression of S100A7. We used qRT-PCR and Western blotting to detect the expression levels of MSX2P1 and S100A7. We down-regulated the expression of MSX2P1 by infecting with lentiviral-vector shRNA. We measured cell proliferation, cell cycle status, and apoptosis by the CCK-8 assay, flow cytometry, and Annexin Ⅴ-FITC/PI staining, respectively. In addition, we used the luciferase reporter gene assay to determine the relationships between MSX2P1 or miR-6731-5p and S100A7, respectively. LncRNA array analysis revealed that 103 lncRNAs were up-regulated and 51 were down-regulated. Furthermore, qRT-PCR showed that the mRNAs levels of MSX2P1 was significantly altered in HaCaT cells treated with IL-22, compared with control cells; and MSX2P1 was mainly in the cytoplasm. Based on the IL-22-stimulated lncRNA-associated ceRNA network, we selected MSX2P1-miR-6731-5p-S100A7 for further study. H&E staining exhibited characteristic features specific to psoriatic lesions. Immunohistochemistry demonstrated significantly increased expression levels of S100A7 in psoriatic lesions, compared with normal skin tissue. We observed a positive correlation between lncRNA-MSX2P1 expression and S100A7 expression. In addition, miR-6731-5p suppressed proliferation, accelerated apoptosis in IL-22-stimulated keratinocytes, and decreased the expressions of S100A7, IL-12β, IL-23, HLA-C, CCHCR1, TNF-α, and NF-κB proteins. Our data demonstrated that MSX2P1 facilitate the progression and growth of IL-22-stimulated keratinocytes by inhibiting miR-6731-5p and activating S100A7. We speculate that the biological network of MSX2P1-miR-6731-5p-S100A7 is a potential novel therapeutic target for the future treatment of psoriasis.
竞争性内源性 RNA（ceRNA）通过竞争共享的 microRNA 来调节 RNA 转录，并在疾病发展中发挥关键作用。银屑病是一种持久的、反复发作的慢性炎症性皮肤病，其特征是角质形成细胞过度增殖。角质形成细胞的反应是由炎症细胞因子（如白细胞介素-22（IL-22））的激活触发的。我们使用 lncRNA 芯片分析来检测用或不用 IL-22 处理的皮肤（HaCaT）细胞中差异表达的 lncRNA。我们使用苏木精和伊红（H＆E）染色来确定皮肤细胞的病理变化，并通过免疫组织化学评估 S100A7 的表达。我们使用 qRT-PCR 和 Western blotting 检测 MSX2P1 和 S100A7 的表达水平。我们通过感染慢病毒载体 shRNA 下调 MSX2P1 的表达。我们通过 CCK-8 测定、流式细胞术和 Annexin Ⅴ-FITC/PI 染色分别测量细胞增殖、细胞周期状态和细胞凋亡。此外，我们使用荧光素酶报告基因测定来确定 MSX2P1 或 miR-6731-5p 与 S100A7 之间的关系。lncRNA 芯片分析显示 103 个 lncRNA 上调，51 个下调。此外，qRT-PCR 显示与对照细胞相比，IL-22 处理的 HaCaT 细胞中 MSX2P1 的 mRNA 水平显着改变；并且 MSX2P1 主要在细胞质中。基于 IL-22 刺激的 lncRNA 相关 ceRNA 网络，我们选择 MSX2P1-miR-6731-5p-S100A7 进行进一步研究。H＆E 染色显示出银屑病病变特有的特征。免疫组织化学显示与正常皮肤组织相比，银屑病病变中 S100A7 的表达水平显着增加。我们观察到 lncRNA-MSX2P1 表达与 S100A7 表达之间存在正相关。此外，miR-6731-5p 抑制增殖，加速 IL-22 刺激的角质形成细胞凋亡，并降低 S100A7、IL-12β、IL-23、HLA-C、CCHCR1、TNF-α 和 NF-κB 蛋白的表达。我们的数据表明，MSX2P1 通过抑制 miR-6731-5p 并激活 S100A7 促进 IL-22 刺激的角质形成细胞的进展和生长。我们推测 MSX2P1-miR-6731-5p-S100A7 的生物学网络是未来治疗银屑病的潜在新治疗靶点。
