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Expression profile and bioinformatics analyses of circular RNAs in keloid and normal dermal fibroblasts.
Experimental Cell Research ( IF 3.3 ) Pub Date : 2020-01-02 , DOI: 10.1016/j.yexcr.2019.111799 Zhibin Zhang 1 , Kaihui Yu 2 , Ougen Liu 1 , Yifeng Xiong 3 , Xinyue Yang 1 , Shuhua Wang 1 , Shulan Zhang 1 , Yueying Feng 4 , Yating Peng 1
Experimental Cell Research ( IF 3.3 ) Pub Date : 2020-01-02 , DOI: 10.1016/j.yexcr.2019.111799 Zhibin Zhang 1 , Kaihui Yu 2 , Ougen Liu 1 , Yifeng Xiong 3 , Xinyue Yang 1 , Shuhua Wang 1 , Shulan Zhang 1 , Yueying Feng 4 , Yating Peng 1
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Increasing evidence indicates that circular RNAs (circRNAs) play a crucial regulatory role in the pathogenesis of multiple diseases. However, no study has examined the potential biological function and expression profile of circRNAs in keloid dermal fibroblasts (KDFs). Therefore, the aim of this study to investigate the expression profile of circRNAs and analyze their role in KDFs. Bioinformatic analyses and high-throughput RNA sequencing technology were applied to explore the expression profile of circRNAs in 3 human KDFs and normal dermal fibroblasts (NDFs). The differentially expressed circRNAs were verified by reverse transcription PCR (RT-PCR), quantitative real-time-PCR (qRT-PCR) and Sanger sequencing. A circRNA-microRNA (miRNA)-mRNA interaction network was created using bioinformatics tools. Hsa_circ_0008259, was selected to confirm its function by qRT-PCR and Western blot. Collectively, 411 circRNAs, of which 206 were upregulated and 205 decreased, were found to be differentially expressed in KDFs and could bind to 2532 miRNA response elements (MREs). GO and KEGG pathways enrichment analyses showed that differentially expressed circRNAs were mainly involved in apoptosis, focal adhesion, PI3K-Akt and metabolic pathway, and may regulate the pathogenesis and development of keloid. Two candidate circRNAs (hsa_circRNA_0008259, hsa_circRNA_0005480) were verified to be significantly reduced in KDFs, and one candidate circRNA (hsa_circRNA_0002198) was significantly elevated in accordance with RNA-Seq data analysis. Overexpression of hsa_circRNA_0008259 inhibited type I and Ⅲ collagen expression. Taken together, our study demonstrates for the first time that circRNAs exhibits differential expression in KDFs, and may be key players in the pathogenesis of keloid, or act as biomarkers of keloid.
中文翻译：
瘢痕loid和正常皮肤成纤维细胞中环状RNA的表达谱和生物信息学分析。
越来越多的证据表明，环状RNA（circRNA）在多种疾病的发病机理中起着至关重要的调节作用。然而，尚无研究检查瘢痕loid皮肤成纤维细胞（KDFs）中circRNA的潜在生物学功能和表达谱。因此，本研究的目的是调查circRNA的表达谱并分析其在KDF中的作用。应用生物信息学分析和高通量RNA测序技术来研究circRNA在3种人类KDF和正常真皮成纤维细胞（NDF）中的表达谱。通过逆转录PCR（RT-PCR），定量实时PCR（qRT-PCR）和Sanger测序验证差异表达的circRNA。使用生物信息学工具创建了circRNA-microRNA（miRNA）-mRNA相互作用网络。Hsa_circ_0008259，通过qRT-PCR和Western印迹来选择其功能以确认其功能。总共发现411个circRNA，其中206个被上调，而205个被减少，它们在KDF中差异表达，并且可以与2532个miRNA响应元件（MRE）结合。GO和KEGG途径的富集分析表明，差异表达的circRNA主要参与细胞凋亡，黏着斑粘附，PI3K-Akt和代谢途径，并可能调节瘢痕loid的发病和发展。证实两个候选circRNA（hsa_circRNA_0008259，hsa_circRNA_0005480）在KDF中显着减少，并且根据RNA-Seq数据分析，一个候选circRNA（hsa_circRNA_0002198）显着升高。hsa_circRNA_0008259的过表达抑制了I型和Ⅲ型胶原蛋白的表达。在一起
更新日期：2020-01-02
中文翻译：
瘢痕loid和正常皮肤成纤维细胞中环状RNA的表达谱和生物信息学分析。
越来越多的证据表明，环状RNA（circRNA）在多种疾病的发病机理中起着至关重要的调节作用。然而，尚无研究检查瘢痕loid皮肤成纤维细胞（KDFs）中circRNA的潜在生物学功能和表达谱。因此，本研究的目的是调查circRNA的表达谱并分析其在KDF中的作用。应用生物信息学分析和高通量RNA测序技术来研究circRNA在3种人类KDF和正常真皮成纤维细胞（NDF）中的表达谱。通过逆转录PCR（RT-PCR），定量实时PCR（qRT-PCR）和Sanger测序验证差异表达的circRNA。使用生物信息学工具创建了circRNA-microRNA（miRNA）-mRNA相互作用网络。Hsa_circ_0008259，通过qRT-PCR和Western印迹来选择其功能以确认其功能。总共发现411个circRNA，其中206个被上调，而205个被减少，它们在KDF中差异表达，并且可以与2532个miRNA响应元件（MRE）结合。GO和KEGG途径的富集分析表明，差异表达的circRNA主要参与细胞凋亡，黏着斑粘附，PI3K-Akt和代谢途径，并可能调节瘢痕loid的发病和发展。证实两个候选circRNA（hsa_circRNA_0008259，hsa_circRNA_0005480）在KDF中显着减少，并且根据RNA-Seq数据分析，一个候选circRNA（hsa_circRNA_0002198）显着升高。hsa_circRNA_0008259的过表达抑制了I型和Ⅲ型胶原蛋白的表达。在一起
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