Gene expression analysis of fibroblasts of systemic sclerosis patients silenced for lncRNA H19X. Gene expression analysis of fibroblasts of systemic sclerosis patients silenced for lncRNA H19X

LncRNA H19X was silienced in dermal fibroblats of systemic sclerosis patients with antisense oligonuclotides. The hypothesis tested in the present study was that H19X is an important factor in the development of TGFb-driven fibrosis. Results provide important information about the role H19X in fibroblasts in particolar on extracellular matrix production and cell cycle regulation. Overall design: Total RNA obtained from dermal fibroblssts of systemic sclerosis patients transfected with antisense oligonucleotide targeting H19X for 120h compared to fibroblasts transfected with scrambled control. H19X silenced fibroblasts and scramble fibroblasts were treated with TGFb for the last 72h of transection.

本研究通过反义寡核苷酸（antisense oligonucleotides）在系统性硬化症（systemic sclerosis）患者的皮肤成纤维细胞（dermal fibroblasts）中沉默了长链非编码RNA（long non-coding RNA，LncRNA）H19X。本研究验证的核心假说为：H19X是转化生长因子β（transforming growth factor-β，TGF-β）介导的纤维化进程中的关键调控因子。本研究结果为阐明H19X在成纤维细胞中的生物学功能提供了重要依据，特别是其在细胞外基质（extracellular matrix）生成与细胞周期调控（cell cycle regulation）过程中的作用。

总体实验设计：将靶向H19X的反义寡核苷酸转染系统性硬化症患者皮肤成纤维细胞，持续120小时；以转染错配对照寡核苷酸（scrambled control）的成纤维细胞作为阴性对照。在转染的最后72小时，分别向H19X沉默组与错配对照组成纤维细胞中加入TGF-β进行处理，随后提取两组细胞的总RNA用于后续分析。