A global transcriptome analysis of human epidermal keratinocytes upon inhibition of lncRNA WAKMAR1

Recent study has revealed that long non-coding RNAs (lncRNAs) perform as important regulators of cellular physiology and pathology, which makes them promising therapeutic and diagnostic entities. We found lncRNA WAKMAR1 is significantly down-regulated in wound-edge keratinocytes from venous ulcer and diabetic foot ulcer compared to the normal wounds. To study the genes regulated by WAKMAR1, we transfected lncRNA GapmeRs into human primary epidermal keratinocytes to inhibit its expression. We performed a global transcriptome analysis of keratinocytes upon inhibition of WAKMAR1 using Affymetrix arrays. We performed a global transcriptome analysis of keratinocytes upon inhibition of WAKMAR1 using Affymetrix arrays. Expression profiling of primary human epidermal keratinocytes transfected with 20nM WAKMAR1 GapmeRs (GapmeR-WAKMAR1) 20nM control oligos (GapmeR-Ctrl) for 24 hours (biological triplicates in each group) was performed using Affymetrix GeneTitan system.

近期研究证实，长链非编码RNA（long non-coding RNAs，lncRNAs）是细胞生理与病理过程的关键调控因子，使其成为极具潜力的治疗与诊断靶点。本研究发现，与正常伤口相比，静脉溃疡与糖尿病足溃疡患者伤口边缘角质形成细胞中的lncRNA WAKMAR1表达显著下调。为探究WAKMAR1调控的靶基因，本研究将lncRNA GapmeRs转染至人原代表皮角质形成细胞以抑制其表达。本研究采用Affymetrix芯片对WAKMAR1被抑制后的角质形成细胞开展全转录组分析。本研究采用Affymetrix芯片对WAKMAR1被抑制后的角质形成细胞开展全转录组分析。本研究采用Affymetrix GeneTitan系统，对转染20nM WAKMAR1 GapmeRs（GapmeR-WAKMAR1）、20nM对照寡核苷酸（GapmeR-Ctrl）并培养24小时的人原代表皮角质形成细胞进行表达谱分析（每组设置3次生物学重复）。