Long non-coding RNA SMILR regulates genes involved in cytokinesis in human vascular smooth muscle cell. Long non-coding RNA SMILR regulates genes involved in cytokinesis in human vascular smooth muscle cell

In response to blood vessel wall injury, aberrant proliferation of vascular smooth muscle cells causes pathological remodelling. The controlling mechanisms involved are, however, not completely understood. We recently showed that the intergenic human, poorly-conserved long non-coding RNA, SMILR, promotes vSMC proliferation. Here, using deep RNA-sequencing of vSMCs with SMILR knockdown or overexpression, we identified an interconnected cell cycle network controlled by SMILR. SMILR inhibition led to vSMC binucleation and mitotic arrest. Overall design: RNA sequencing was performed on primary Human Saphenous Vein Smooth Muscle Cells (HSVSMCs), stimulated with IL1a (10 ng/mL) and PDGF-ββ (20 ng/mL), exposed to either SMILR depletion via siRNA (siSMILR) or overexpression via lentivirus (SMILR_LV). Respective control corresponds to siCONTROL or empty lentivirus. Each treatment was done in triplicates.

当血管壁遭受损伤时，血管平滑肌细胞（vascular smooth muscle cells, VSMC）的异常增殖会引发病理性血管重构，然而其背后的调控机制尚未完全阐明。我们此前的研究证实，人类基因间区保守性极差的长链非编码RNA（long non-coding RNA, lncRNA）SMILR可促进VSMC增殖。本研究中，我们对SMILR敲低或过表达的VSMC开展深度RNA测序，鉴定出了SMILR调控的相互关联的细胞周期网络。抑制SMILR可导致VSMC发生双核化及有丝分裂阻滞。
实验设计概述：本实验以人隐静脉平滑肌原代细胞（Human Saphenous Vein Smooth Muscle Cells, HSVSMCs）为研究对象，先用浓度为10 ng/mL的IL-1α及20 ng/mL的PDGF-ββ进行刺激，随后分别通过小干扰RNA（small interfering RNA, siRNA）介导SMILR敲低（siSMILR组）或慢病毒介导SMILR过表达（SMILR_LV组）进行处理；相应对照组分别为siCONTROL对照组及空载慢病毒对照组。每组实验均设置三次生物学重复。