MiR-548a-3p Promotes Keratinocyte Proliferation Targeting PPP3R1 after Being Induced by IL-22
MiR-548a-3p Promotes Keratinocyte Proliferation Targeting PPP3R1 after Being Induced by IL-22
复制标题
IL-22 诱导后 MiR-548a-3p 促进靶向 PPP3R1 的角质形成细胞增殖
DOI：
10.1007/s10753-017-0705-3
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发表时间：
2018-03-01
期刊：
影响因子：
5.1
通讯作者：
Sun, Qing
中科院分区：
文献类型：
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作者：
Zhao, Xintong;Li, Ronghua;Sun, Qing
银屑病是一种免疫介导的慢性皮肤病，T细胞起主要作用，多种炎性细胞因子通过启动角质形成细胞增殖参与其发病机制。白细胞介素-22（IL-22）是IL-10家族的一种细胞因子，在银屑病的发病和发展中起重要作用。为了确定microRNA（miR）-548a-3p的靶点并研究其在用IL-22处理人角质形成细胞（HaCaT）后角质形成细胞增殖中的作用，我们使用定量逆转录酶PCR来测量miR-548 a-3 p在用IL-22刺激的HaCaT细胞和银屑病病变中的表达，采用CCK-8法检测miR-548 a-3 p在HaCaT细胞中的生物学功能。采用荧光素酶报告基因分析法确定miR-548 a-3 p的靶基因。免疫组化和Western blot检测目的基因。结果显示，miR-548 a-3 p在用IL-22处理的HaCaT细胞和银屑病病变中均显著上调。miR-548 a-3 p的过表达可促进HaCaT细胞的增殖。荧光素酶在PPP 3R 1（编码钙调磷酸酶的基因）的3 'UTR中突变。免疫组化和Western blot结果显示，PPP 3R 1在银屑病皮损和HaCaT细胞中的表达均降低。总之，miR-548 a-3 p的表达在IL-22介导的角质形成细胞增殖性疾病如银屑病中上调。miR-548 a-3 p对角质形成细胞增殖的影响可能通过靶向PPP 3R 1来实现，调节性T细胞可能参与银屑病的发病机制。
AbstractPsoriasis is an immune-mediated chronic skin disorder where T cells play a main role, and numerous inflammatory cytokines are implicated in its pathogenesis by initiating keratinocyte proliferation. Interleukin-22 (IL-22), an IL-10 family cytokines, is critical in the pathogenesis and development of psoriasis. To determine the target of microRNA (miR) -548a-3p and investigate its role in keratinocyte proliferation after treating human keratinocytes (HaCaT) with IL-22, we used quantitative reverse transcriptase PCR to measure the expression of miR-548a-3p in both HaCaT cells stimulated with IL-22 and psoriatic lesions, and then detected the biological function of miR-548a-3p in HaCaT cells by performing Counting Kit-8 (CCK-8) assays. Luciferase reporter assay was conducted to determine the target gene of miR-548a-3p. Immunohistochemistry and Western blot were performed to verify the target gene. Results showed that miR-548a-3p was significantly upregulated both in HaCaT cells treated with IL-22 and psoriatic lesions. The over expression of miR-548a-3p could promote the proliferation of HaCaT cells. Luciferase was mutated in the 3’UTR of PPP3R1, a gene coding Calcineurin. Immunohistochemistry and Western blot demonstrated that the expression of PPP3R1 decreased respectively in psoriatic lesions and HaCaT cells. In conclusion, the expression of miR-548a-3p is upregulated in IL-22 mediated keratinocyte proliferative disorder like psoriasis. The impact of miR-548a-3p on keratinocyte proliferation may be implemented by targeting PPP3R1 and T regulatory cells may be involved in the pathogenesis of psoriasis.