Small Non-Coding RNAome of Ageing Chondrocytes
Small Non-Coding RNAome of Ageing Chondrocytes
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DOI：
10.3390/ijms21165675
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发表时间：
2020-08-01
影响因子：
5.6
通讯作者：
Peffers, Mandy J.
中科院分区：
文献类型：
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作者：
Balaskas, Panagiotis;Green, Jonathan A.;Peffers, Mandy J.
老化是诱发软骨骨关节炎的主要危险因素。然而，关于衰老对小的非编码RNA（sncRNA）表达的影响的研究很少。从肉眼正常的马掌指关节的年轻和年老软骨细胞中提取RNA，并进行小RNA测序（RNA-seq）。使用包DESeq 2在R中进行差异表达分析。对于转移RNA（tRNA）片段分析，使用Bowtie 2 2.2.5版将tRNA读数与马tRNA序列进行比对。在马软骨细胞的扩展队列中，使用实时定量聚合酶链反应（qRT-PCR）验证选定的microRNA（miRNA或miR）和小核仁RNA（snoRNA）结果。在低级别和高级别OA人软骨组织中进一步研究了tRNA片段。总共有83个sncRNA在年轻和年老的马软骨细胞之间差异表达，包括miRNA、snoRNA、小核RNA（snRNA）和tRNA。qRT-PCR分析证实了结果。tRNA片段分析显示，tRNA半体（tiRNA）、tiRNA-5035-GluCTC和tiRNA-5031-GluCTC-1在高度OA人软骨和老年马软骨细胞中均减少。这是第一次，我们已经测量了马软骨细胞中sncRNA表达的衰老效应。在许多不同的sncRNA种类中检测到变化。这项研究支持sncRNA在老化软骨中的作用及其在年龄相关软骨疾病中的潜在参与。
Ageing is a leading risk factor predisposing cartilage to osteoarthritis. However, little research has been conducted on the effect of ageing on the expression of small non-coding RNAs (sncRNAs). RNA from young and old chondrocytes from macroscopically normal equine metacarpophalangeal joints was extracted and subjected to small RNA sequencing (RNA-seq). Differential expression analysis was performed in R using package DESeq2. For transfer RNA (tRNA) fragment analysis, tRNA reads were aligned to horse tRNA sequences using Bowtie2 version 2.2.5. Selected microRNA (miRNAs or miRs) and small nucleolar RNA (snoRNA) findings were validated using real-time quantitative Polymerase Chain Reaction (qRT-PCR) in an extended cohort of equine chondrocytes. tRNA fragments were further investigated in low- and high-grade OA human cartilage tissue. In total, 83 sncRNAs were differentially expressed between young and old equine chondrocytes, including miRNAs, snoRNAs, small nuclear RNAs (snRNAs), and tRNAs. qRT-PCR analysis confirmed findings. tRNA fragment analysis revealed that tRNA halves (tiRNAs), tiRNA-5035-GluCTC and tiRNA-5031-GluCTC-1 were reduced in both high grade OA human cartilage and old equine chondrocytes. For the first time, we have measured the effect of ageing on the expression of sncRNAs in equine chondrocytes. Changes were detected in a number of different sncRNA species. This study supports a role for sncRNAs in ageing cartilage and their potential involvement in age-related cartilage diseases.