IL-22-induced miR-122-5p promotes keratinocyte proliferation by targeting Sprouty2
IL-22-induced miR-122-5p promotes keratinocyte proliferation by targeting Sprouty2
复制标题
IL-22 诱导的 miR-122-5p 通过靶向 Sprouty2 促进角质形成细胞增殖
DOI：
10.1111/exd.13270
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发表时间：
2017-04-01
影响因子：
3.6
通讯作者：
Sun, Qing
中科院分区：
文献类型：
--
作者：
Jiang, Meng;Ma, Weiyuan;Sun, Qing
银屑病是一种常见的炎症性皮肤病，但确切的发病机制很大程度上尚不清楚。白介素22(IL-22)在银屑病发病机制中与角质形成细胞相互作用，在T细胞介导的免疫反应中发挥重要作用。在这里，我们使用miRNA和mRNA微阵列展示了在IL-22刺激的HaCaT细胞中差异表达的miRNAs及其潜在靶点。我们发现HaCaT细胞中有20个miRNAs发生了显着变化(超过两倍)，并用定量逆转录聚合酶链式反应(qRT-PCR)验证了结果。我们发现在IL-22刺激的HaCaT细胞和银屑病皮损中miR-122-5p表达上调。然后，我们旨在研究miR-122-5p在角质形成细胞中的生物学作用及其可能的机制。CCK-8检测结果表明，在角质形成细胞中过表达miR-122-5p可促进角质形成细胞的增殖，而抑制内源性miR-122-5p可抑制角质形成细胞的增殖。根据芯片分析，我们推测Sprouty2(Sprouty2，Sprouty2)是miR-122-5p的直接靶基因，它是细胞外信号调节激酶/丝裂原活化蛋白激酶信号通路的负调控因子。免疫组织化学检测发现，SPRY2在细胞质中的表达主要定位于表皮的基底层和基底层，且在银屑病组织中的表达明显低于正常对照组。HaCaT细胞荧光素酶报告基因和Western印迹分析表明，SPRY2是miR-122-5p的直接靶基因。结论：IL-22诱导的miR-122-5p可能通过下调SPRY2的表达促进角质形成细胞的增殖，从而在银屑病的发病机制中发挥重要作用。
Psoriasis is a common inflammatory skin disease, but the exact pathogenesis is largely unknown. Interleukin-22 (IL-22) has demonstrated its vital role in T-cell-mediated immune response by interacting with keratinocytes in the pathogenesis of psoriasis. Here, we showed the differentially expressed miRNAs and their potential targets in HaCaT cells stimulated by IL-22 using miRNA and mRNA microarrays. We revealed a total of 20 significantly changed (more than twofold) miRNAs in HaCaT cells and validated the results with quantitative reverse transcriptase PCR (qRT-PCR). We demonstrated that miR-122-5p was up-regulated both in HaCaT cells stimulated by IL-22 and in psoriatic lesions. Then, we aimed to investigate the biological roles and potential mechanism of miR-122-5p in keratinocytes. As a result, CCK-8 assay indicated that overexpression of miR-122-5p in keratinocytes promoted proliferation and conversely inhibition of endogenous miR-122-5p suppressed proliferation. According to the microarray analysis, we assumed that Sprouty2 (Spry2), a negative regulator of extracellular signal regulated kinase/mitogen-activated protein kinase signalling pathway, was a direct target gene of miR-122-5p. We found that the staining of Spry2 in cytoplasm was mainly localized in both basal and suprabasal layers of epidermis and showed a markedly decreased expression in psoriasis than in normal control by immunohistochemistry. Luciferase reporter and Western blot assays in HaCaT cells demonstrated that Spry2 was a direct target gene of miR-122-5p. In conclusion, IL-22-induced miR-122-5p promotes keratinocyte proliferation possibly by downregulating the expression of Spry2 thus playing important roles in the pathogenesis of psoriasis.