circCOL3A1-859267 regulates type I collagen expression by sponging miR-29c in human dermal fibroblasts
circCOL3A1-859267 regulates type I collagen expression by sponging miR-29c in human dermal fibroblasts
复制标题
circCOL3A1-859267 通过海绵 miR-29c 在人真皮成纤维细胞中调节 I 型胶原表达
DOI：
10.1684/ejd.2018.3397
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发表时间：
2018-09-01
影响因子：
2.5
通讯作者：
Lai, Wei
中科院分区：
文献类型：
--
作者：
Peng, Yating;Song, Xiaojing;Lai, Wei
背景基因COL1A1和COL1A2分别编码I型胶原蛋白的pro-α1和pro-α2链，I型胶原蛋白是皮肤真皮的主要成分之一。我们之前已经证明，环状RNA（一类最近发现的非编码RNA，通过像海绵一样隔离miRNA来表现出调节效力），我们称之为circCOL3A1-859267，在皮肤真皮中下调并调节I型胶原蛋白的表达。暴露于 UVA 的人真皮成纤维细胞 (HDF)。然而，在UVA照射的HDF中circCOL3A1-859267介导的胶原表达的精确机制仍不清楚。目的阐明circCOL3A1-859267介导的I型胶原表达调节机制。材料与方法我们最初基于生物信息学方法和双荧光素酶报告基因分析预测了circCOL3A1-859267上的miRNA结合位点用于测定 HEK 293 细胞中 miRNA 与 circCOL3A1-859267 的结合。在 HDF 中进一步研究了 UVA 照射对 miRNA 表达以及 circCOL3A1-859267 介导的 I 型胶原蛋白表达的影响。 结果 miR-29a、miR-29b、miR-29c、miR-767 和 miR-133a 预计会结合 circCOL3A1-859267 和 COL1A1/COL1A2，然而，仅结合基于双荧光素酶报告基因测定，miR-29c 显示与 circCOL3A1-859267 结合。在 UVA 暴露的 HDF 中，只有 miR-29c 上调。最后，转染靶向 circCOL3A1-859267 或 miR-29c 模拟物的小干扰 RNA 抑制 HDF 中 I 型胶原蛋白的表达；使用基于慢病毒的表达系统过表达 circCOL3A1-859267，可恢复 miR-29c 模拟物诱导的下调。结论我们的结果表明，circCOL3A1-859267 通过在 HDF 中海绵和隔离 miR-29c 来调节 I 型胶原表达。
BackgroundThe genes COL1A1 and COL1A2 encode the pro-alpha 1 and pro-alpha 2 chains of type I collagen, respectively, which is one of the main components of skin dermis.We have previously demonstrated that a circularRNA(a class of recently identified non-codingRNAs that exhibit regulatory potency by sequestering miRNAs like a sponge), which we termed circCOL3A1-859267, is downregulated and regulates type I collagen expression in UVA-exposed human dermal fibroblasts (HDFs). However, the precise mechanisms of circCOL3A1-859267-mediated collagen expression in UVA-irradiated HDFs remain unclear.ObjectivesTo elucidate the mechanism of circCOL3A1-859267-mediated regulation of type I collagen expression.Materials & methodsWe initially predicted miRNA binding sites on circCOL3A1-859267 based on a bioinformatic method, and a dual luciferase reporter assay was used to determine miRNA binding to circCOL3A1-859267 in HEK 293 cells. The effect of UVA irradiation on the expression of miRNAs as well as circCOL3A1-859267-mediated type I collagen expression was further investigated in HDFs.ResultsmiR-29a, miR-29b, miR-29c, miR-767, and miR-133a were predicted to bind both circCOL3A1-859267 and COL1A1/COL1A2, however, only miR-29c was shown to bind to circCOL3A1-859267 based on the dual luciferase reporter assay. In UVA-exposed HDFs, only miR-29c was upregulated. Finally, transfection of a small interfering RNA targeting circCOL3A1-859267 or miR-29c mimic suppressed the expression of type I collagen in HDFs; the miR-29c mimic-induced down-regulation was restored via overexpression of circCOL3A1-859267 using a lentiviral-based expression system.ConclusionOur results indicate that circCOL3A1-859267 regulates type I collagen expression by sponging and sequestering miR-29c in HDFs.