一种人类长链非编码RNA ALT1通过ACE2和细胞周期蛋白D1途径调控血管内皮细胞的细胞周期。
A Human Long Non-Coding RNA ALT1 Controls the Cell Cycle of Vascular Endothelial Cells Via ACE2 and Cyclin D1 Pathway.
作者信息
Li Wen, Wang Rui, Ma Jie-Yi, Wang Mian, Cui Jin, Wu Wei-Bin, Liu Rui-Ming, Zhang Chun-Xiang, Li Wen, Wang Shen-Ming
机构信息
Division of Vascular Surgery, Guangdong Key Engineering Laboratory for Diagnosis and Treatment of Vascular Disease, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China.
Laboratory of General Surgery, Guangdong Key Engineering Laboratory for Diagnosis and Treatment of Vascular Disease, First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China.
出版信息
Cell Physiol Biochem. 2017;43(3):1152-1167. doi: 10.1159/000481756. Epub 2017 Oct 5.
BACKGROUND/AIMS: ALT1 is a novel long non-coding RNA derived from the alternatively spliced transcript of the deleted in lymphocytic leukemia 2 (DLEU2). To date, ALT1 biological roles in human vascular endothelial cells have not been reported.
METHODS
ALT1 was knocked down by siRNAs. Cell proliferation was analyzed by cck-8. The existence and sequence of human ALT1 were identified by 3' rapid amplification of cDNA ends. The interaction between lncRNA and proteins was analyzed by RNA-Protein pull down assay, RNA immunoprecipitation, and mass spectrometry analysis.
RESULTS
ALT1 was expressed in human umbilical vein endothelial cells (HUVECs). The expression of ALT1 was significantly downregulated in contact-inhibited HUVECs and in hypoxia-induced, growth-arrested HUVECs. Knocking down of ALT1 inhibited the proliferation of HUVECs by G0/G1 cell cycle arrest. We observed that angiotensin converting enzyme Ⅱ(ACE2) was a direct target gene of ALT1. Knocking-down of ALT1 or its target gene ACE2 could efficiently decrease the expression of cyclin D1 via the enhanced ubiquitination and degradation, in which HIF-1α and protein von Hippel-Lindau (pVHL) might be involved.
CONCLUSION
The results suggested the human long non-coding RNA ALT1 is a novel regulator for cell cycle of HUVECs via ACE2 and cyclin D1 pathway.
背景/目的：ALT1是一种新型长链非编码RNA，源自淋巴细胞白血病2（DLEU2）基因可变剪接转录本。迄今为止，尚未见ALT1在人血管内皮细胞中的生物学作用报道。
方法
利用小干扰RNA（siRNA）敲低ALT1表达。采用细胞计数试剂盒-8（cck-8）分析细胞增殖情况。通过3' cDNA末端快速扩增法鉴定人ALT1的存在及序列。运用RNA-蛋白质下拉实验、RNA免疫沉淀和质谱分析检测长链非编码RNA（lncRNA）与蛋白质之间的相互作用。
结果
ALT1在人脐静脉内皮细胞（HUVECs）中表达。在接触抑制的HUVECs以及缺氧诱导生长停滞的HUVECs中，ALT1表达显著下调。敲低ALT1通过使细胞周期停滞在G0/G1期抑制HUVECs增殖。我们观察到血管紧张素转换酶Ⅱ（ACE2）是ALT1的直接靶基因。敲低ALT1或其靶基因ACE2可通过增强泛素化和降解有效降低细胞周期蛋白D1的表达，其中可能涉及缺氧诱导因子-1α（HIF-1α）和冯·希佩尔-林道蛋白（pVHL）。
结论
结果表明人长链非编码RNA ALT1通过ACE2和细胞周期蛋白D1途径对HUVECs细胞周期具有新型调控作用。
Zotero 插件