Zha Weifeng, Guo Bo, Chen Shuyue, Lu Junwei, Shan Yunyun
Department of Dermatology, Third People's Hospital of Hangzhou, Hangzhou, Zhejiang 310009, P.R. China.
Department of Dermatology, Tongxiang Dermatosis Prevention Institute, Tongxiang, Zhejiang 314500, P.R. China.
Exp Ther Med. 2021 May;21(5):450. doi: 10.3892/etm.2021.9878. Epub 2021 Mar 1.
Psoriasis is a T-cell-mediated inflammatory skin disease that is characterized by excessive keratinocyte proliferation and persistent skin inflammation. Accumulating evidence suggests that long non-coding RNAs (lncRNAs) are dysregulated in a number of inflammatory conditions. In the present study, an psoriasis cell model was established. Human HaCaT keratinocytes were activated using the inflammatory factor IL-22. Briefly, HaCaT cells were starved in serum-free DMEM for 24 h and then stimulated with 100 ng/ml IL-22 in serum-free DMEM for 24 h. Previous research indicated that HOX transcript antisense RNA (HOTAIR) may participate in the development of psoriasis. First, reverse transcription-quantitative PCR (RT-qPCR) analysis was performed to detect HOTAIR expression. The results indicated that HOTAIR expression was reduced in IL-22-stimulated HaCaT cells. Subsequently, a dual-luciferase reporter assay was performed to verify the binding site between HOTAIR and microRNA (miR)-126. The RT-qPCR results indicated that miR-126 expression was increased in IL-22-stimulated HaCaT cells. Moreover, the effects of HOTAIR and miR-126 on IL-22-stimulated HaCaT cell proliferation and apoptosis were assessed. HaCaT cells were transfected with control-plasmid, HOTAIR-plasmid, HOTAIR-plasmid + mimic control or HOTAIR-plasmid + miR-126 mimic for 24 h. At 24 h post-transfection, the cells were stimulated with 100 ng/ml IL-22 for 24 h and experiments were conducted. IL-22 induced cell proliferation and suppressed apoptosis. However, HOTAIR-plasmid inhibited cell viability and induced apoptosis in IL-22-stimulated HaCaT cells. In addition, the western blotting results indicated that HOTAIR-plasmid increased cleaved caspase-3 expression and the cleaved caspase-3/caspase-3 ratio, whereas the HOTAIR-plasmid-mediated effects were reversed by miR-126 mimic. Collectively, the results of the present study demonstrated that the lncRNA-HOTAIR/miR-126 axis may be implicated in the regulation of psoriasis progression and may serve as a potential therapeutic target for psoriasis.
银屑病是一种由T细胞介导的炎症性皮肤病，其特征为角质形成细胞过度增殖和持续性皮肤炎症。越来越多的证据表明，长链非编码RNA（lncRNA）在多种炎症性疾病中表达失调。在本研究中，建立了银屑病细胞模型。使用炎性因子IL-22激活人HaCaT角质形成细胞。简要地说，将HaCaT细胞在无血清DMEM中饥饿24小时，然后在无血清DMEM中用100 ng/ml IL-22刺激24小时。先前的研究表明，HOX转录本反义RNA（HOTAIR）可能参与银屑病的发展。首先，进行逆转录定量PCR（RT-qPCR）分析以检测HOTAIR的表达。结果表明，在IL-22刺激的HaCaT细胞中HOTAIR表达降低。随后，进行双荧光素酶报告基因检测以验证HOTAIR与微小RNA（miR）-126之间的结合位点。RT-qPCR结果表明，在IL-22刺激的HaCaT细胞中miR-126表达增加。此外，评估了HOTAIR和miR-126对IL-22刺激的HaCaT细胞增殖和凋亡的影响。将HaCaT细胞用对照质粒、HOTAIR质粒、HOTAIR质粒+模拟对照或HOTAIR质粒+miR-126模拟物转染24小时。转染后24小时，用100 ng/ml IL-22刺激细胞24小时并进行实验。IL-22诱导细胞增殖并抑制凋亡。然而，HOTAIR质粒抑制IL-22刺激的HaCaT细胞的活力并诱导凋亡。此外，蛋白质印迹结果表明，HOTAIR质粒增加了裂解的caspase-3表达以及裂解的caspase-3/caspase-3比值，而miR-126模拟物可逆转HOTAIR质粒介导的作用。总体而言，本研究结果表明lncRNA-HOTAIR/miR-126轴可能参与银屑病进展的调控，并可能作为银屑病的潜在治疗靶点。
