Downregulated miR-27b promotes keratinocyte proliferation by targeting PLK2 in oral lichen planus
Downregulated miR-27b promotes keratinocyte proliferation by targeting PLK2 in oral lichen planus
复制标题
下调miR-27b通过靶向口腔扁平苔藓中的PLK2促进角质形成细胞增殖
DOI：
10.1111/jop.12826
复制
发表时间：
2019-04-01
影响因子：
3.3
通讯作者：
Tang, Guoyao
中科院分区：
文献类型：
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作者：
Chen, Junjun;Du, Guanhuan;Tang, Guoyao
背景最近发现microRNA-27 b（miR-27 b）在口腔扁平苔藓（OLP）中显著下调。然而，miR-27 b在OLP中功能的证据仍然有限。方法采用实时荧光定量聚合酶链反应（qRT-PCR）检测miR-27 b在口腔扁平苔藓中的表达。在功能上，然后使用miR-27 b模拟物/抑制剂进行功能获得/丧失研究，以研究人口腔角质形成细胞（HOK）中的细胞生长。从机制上讲，随后的miRNA靶分析，包括starBase数据库分析和荧光素酶报告基因测定进行预测和验证的直接目标，分别。此外，进行miR-27 b靶点的过表达/敲低测定以研究其功能意义，并分别使用qRT-PCR和蛋白质印迹法评估miR-27 b mRNA和蛋白质水平的靶点。结果OLP组织中MicroRNA-27 b表达明显低于正常对照组织。生物信息学预测波罗样激酶2（PLK 2）可能是miR-27 b的潜在靶点，而荧光素酶报告基因分析结果显示miR-27 b直接抑制plk 2 -3 '非翻译区。此外，功能分析表明，下调miR-27 b导致HOK细胞生长增加，相应地，PLK 2过表达促进HOK增殖。结论miR-27 b和PLK 2在OLP组织中存在异常表达。miR-27 b的减少可能通过增加HOKs中PLK 2的水平来诱导细胞增殖，这为OLP发展的潜在机制提供了新的视角。
Background MicroRNA-27b (miR-27b) was recently found to be significantly downregulated in oral lichen planus (OLP). However, evidence of the function of miR-27b in OLP remains limited. Methods Initially, miR-27b expression in OLP was verified using the quantitative real-time polymerase chain reaction (qRT-PCR). Functionally, gain-/loss-of-function studies were then conducted using miR-27b mimics/inhibitor to investigate cell growth in human oral keratinocytes (HOKs). Mechanistically, subsequent miRNA target analyses including a starBase database analysis and a luciferase reporter assay were performed to predict and validate the direct target, respectively. In addition, overexpression/knockdown assays of target(s) of miR-27b were performed to investigate its functional significance and qRT-PCR and western blotting were used to evaluate the target(s) of miR-27b mRNA and protein levels, respectively. Results MicroRNA-27b was significantly downregulated in OLP tissues when compared with healthy control tissues. Bioinformatics predicted that Polo Like Kinase 2 (PLK2) might be a potential target of miR-27b, while the luciferase reporter assay results showed the direct inhibition of the plk2-3 ' untranslated region by miR-27b. Moreover, functional analysis indicated that downregulated miR-27b caused an increase in cell growth in HOKs, and correspondingly, overexpression of PLK2 promoted HOK proliferation. Conclusions There were aberrant expressions of miR-27b and PLK2 in OLP tissues. Decreased miR-27b may have induced cell proliferation by increasing the levels of PLK2 in HOKs, which provides a new perspective into the potential mechanisms underlying OLP development.