Effect of microRNA-27b-3p mimic treatment on human osteoarthritis fibroblast-like synoviocyte gene expression profiles

OBJECTIVE: To identify the changes in gene expression elicited by miR-27b-3p mimic transfection of human osteoarthritis (OA) primary fibroblast-like synoviocytes (FLS) obtained from patients undergoing knee arthroplasty. Cells were cultured, and treated for 48 h with miR-27b-3p mimic or Cel-miR-39-3p control mimic. Total RNA was then extracted, and mRNA libraries were prepared using Illumina's TruSEQ stranded total RNA library preparation kit, pooled together, and sequenced on Illumina's NextSeq550. Differential expression analysis was performed on the RNA-SEQ data obtained from 6 prepared libraries, where 3 were treated with the miR-27b-3p mimic and 3 with Cel-miR-39-3p control mimic. Subsequent pathway analysis identified the upregulation of ECM-related genes and pathways suggesting that miR-27b-3p is involved in the modulation of synovial ECM production.

研究目的：明确转染miR-27b-3p模拟物（miR-27b-3p mimic）对从接受膝关节置换术患者体内分离的人骨关节炎（osteoarthritis, OA）原代成纤维样滑膜细胞（fibroblast-like synoviocytes, FLS）的基因表达产生的影响。将细胞进行培养后，分别用miR-27b-3p模拟物或Cel-miR-39-3p对照模拟物（Cel-miR-39-3p control mimic）处理48小时。随后提取总RNA，使用Illumina的TruSEQ链特异性总RNA建库试剂盒构建mRNA文库，将文库混合后在Illumina NextSeq550平台上进行测序。对6个制备完成的文库的RNA测序数据开展差异表达分析，其中3个文库经miR-27b-3p模拟物处理，剩余3个经Cel-miR-39-3p对照模拟物处理。后续通路分析结果显示，细胞外基质（extracellular matrix, ECM）相关基因及通路出现上调，提示miR-27b-3p参与调控滑膜细胞外基质的生成。