Feng Shike, Wang Lin, Liu Wang, Zhong Yan, Xu Shijun
Department of Dermatology, The First People's Hospital of Zigong, Zigong, China.
Department of Dermatology, The People's Hospital of Pengzhou, Chengdu, China.
J Clin Lab Anal. 2018 Nov;32(9):e22588. doi: 10.1002/jcla.22588. Epub 2018 Jun 26.
This study aimed to investigate the miR-126 expression in lesional skin and its correlation with clinical features in psoriasis patients and to explore the effect of upregulated miR-126 on cells' proliferation, apoptosis, and inflammation in human keratinocytes.
A total of 102 psoriasis patients were consecutively enrolled in this study. MiR-126 expressions in lesional skin and paired nonlesional skin were detected by quantitative polymerase chain reaction (qPCR). Human keratinocytes (HaCaT cells) were transfected with miR-126 mimic plasmids and blank mimic plasmid. Cell Counting Kit-8 and annexin V/propidium iodide assays were performed to assess the cells' proliferation and apoptosis, and protein levels of apoptotic markers (cleaved caspase-3 [C-caspase-3] and B-cell lymphoma-2 [Bcl-2]) were detected by Western blot assay. Inflammatory cytokines mRNA and protein levels were detected by qPCR and Western blot assays, respectively.
MiR-126 expression was upregulated in lesional skin tissue compared with paired nonlesional skin tissue, and its expression positively associated with Psoriasis Area and Severity Index score in psoriasis patients. MiR-126 expression was increased in miR-126 mimic group compared with negative control (NC) mimic group after plasmids transfection into HaCaT cells, and cells' proliferation was enhanced while cells' apoptosis rate was reduced in miR-126 mimic group than NC mimic group. Protein expressions of C-caspase and Bcl-2 also indicated miR-126 mimic decreased the cells' apoptosis. In addition, miR-126 mimic increased TNF-α, IFN-γ, IL-17A, and IL-22 expressions while decreased IL-10 expression.
In conclusion, miR-126 correlates with elevated risk and increased disease severity in psoriasis patients, and upregulation of miR-126 promotes cells' proliferation and inflammation while inhibits cells' apoptosis in keratinocytes.
本研究旨在调查银屑病患者皮损组织中miR-126的表达及其与临床特征的相关性,并探讨上调miR-126对人角质形成细胞增殖、凋亡和炎症的影响。
本研究连续纳入102例银屑病患者。采用定量聚合酶链反应(qPCR)检测皮损组织及配对的非皮损组织中miR-126的表达。将miR-126模拟质粒和空白模拟质粒转染人角质形成细胞(HaCaT细胞)。采用细胞计数试剂盒-8和膜联蛋白V/碘化丙啶检测法评估细胞增殖和凋亡,并通过蛋白质免疫印迹法检测凋亡标志物(裂解的半胱天冬酶-3 [C-caspase-3]和B细胞淋巴瘤-2 [Bcl-2])的蛋白水平。分别通过qPCR和蛋白质免疫印迹法检测炎性细胞因子的mRNA和蛋白水平。
与配对的非皮损组织相比,皮损组织中miR-126表达上调,且其表达与银屑病患者的银屑病面积和严重程度指数评分呈正相关。将质粒转染至HaCaT细胞后,miR-126模拟组的miR-126表达高于阴性对照(NC)模拟组,且miR-126模拟组的细胞增殖增强,细胞凋亡率低于NC模拟组。C-caspase和Bcl-2的蛋白表达也表明miR-126模拟物降低了细胞凋亡。此外,miR-126模拟物增加了TNF-α、IFN-γ、IL-17A和IL-22的表达,同时降低了IL-10的表达。
总之,miR-126与银屑病患者的患病风险升高和疾病严重程度增加相关,上调miR-126可促进角质形成细胞的增殖和炎症,同时抑制细胞凋亡。