LncRNA RASGRF2-AS1调控HUVECs的增殖、凋亡和血管生成

IF 0.9 Q4 GENETICS & HEREDITY

Gene Reports Pub Date : 2025-08-12 DOI:10.1016/j.genrep.2025.102319

Cairong Liu , Yunyan Liu , Yijie Li , Lin Huang

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引用次数: 0

摘要

氧化低密度脂蛋白（oxLDL）是动脉粥样硬化发展的关键因素，在血管内皮功能障碍的发生和发展中起着促炎介质的重要作用。根据先前的一项研究，RASGRF2-AS1被发现是oxLDL刺激后人脐静脉内皮细胞（HUVECs）中最明显下调的长链非编码rna之一。迄今为止，RASGRF2-AS1的生物学作用尚未报道。因此，我们开始探索RASGRF2-AS1在huvec中的功能。方法采用实时荧光定量PCR （qRT-PCR）检测HUVECs中srasgrf2 - as1的表达。为了沉默RASGRF2-AS1，使用了siRNA和慢病毒介导的shRNA。细胞计数试剂盒-8 （CCK-8）检测细胞增殖情况。然后，我们用annexin V/PI染色检测RASGRF2-AS1敲低后的细胞凋亡和细胞周期分布。western blot检测微管相关蛋白1轻链3 β (MAP1LC3B)和sequestosome 1 （SQSTM1/p62）表达水平。此外，预测与RASGRF2-AS1相互作用的候选蛋白通过RNA下拉试验和质谱测定。结果rasgrf2 - as1在huvec中高表达。用siRNA和shRNA下调RASGRF2-AS1表达后，G0/G1细胞周期阻滞增加，抑制HUVEC增殖。下调RASGRF2-AS1也可促进huvec细胞凋亡，抑制小管形成。此外，western blot结果显示，RASGRF2-AS1敲低可降低p62表达，增加MAP1LC3B表达。此外，RNA pull-down实验确定了几种共沉淀蛋白作为RASGRF2-AS1的潜在相互作用物。这些候选物包括S100-A9、ZN598、NRROS、ZMYM5、IF4A1、PDIP3和PLCB4。结论srasgrf2 - as1是参与调节HUVECs增殖、凋亡和血管生成能力的新型关键lncRNA。因此，RASGRF2-AS1可能是治疗动脉硬化闭塞症的一个有希望的靶点。

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LncRNA RASGRF2-AS1 regulates proliferation, apoptosis, and angiogenesis of HUVECs

Background

Oxidized low-density lipoprotein (oxLDL) is a key contributor to the development of atherosclerosis and plays a crucial role as a proinflammatory mediator in the onset and progression of vascular endothelial dysfunction. According to a previous study, RASGRF2-AS1 was found to be one of the most notably downregulated long noncoding RNAs in human umbilical vein endothelial cells (HUVECs) following oxLDL stimulation. To date, the biological roles of RASGRF2-AS1 have not been reported. Therefore, we set out to explore the function of RASGRF2-AS1 in HUVECs.

Methods

RASGRF2-AS1 expression in HUVECs was measured using quantitative real-time PCR (qRT-PCR). To silence RASGRF2-AS1, both siRNA and lentivirus-mediated shRNA were utilized. The Cell Counting Kit-8 (CCK-8) assay was employed to evaluate cell proliferation. Then, we used annexin V/PI staining to determine cell apoptosis and cell cycle distribution after RASGRF2-AS1 knockdown. Microtubule-associated protein 1 light chain 3 β (MAP1LC3B) and sequestosome 1 (SQSTM1/p62) expression levels were also measured by western blot. Furthermore, candidate proteins predicted to interact with RASGRF2-AS1 were determined by RNA pull-down assays and mass spectrometry.

Results

RASGRF2-AS1 was highly expressed in HUVECs. After RASGRF2-AS1 expression was downregulated with siRNA and shRNA, G0/G1 cell cycle arrest increased, inhibiting HUVEC proliferation. Downregulating RASGRF2-AS1 also promoted apoptosis and suppressed tube formation in HUVECs. In addition, the western blot results indicated that RASGRF2-AS1 knockdown decreased p62 expression and increased MAP1LC3B expression. Furthermore, RNA pull-down assays identified several co-precipitating proteins as potential interactors of RASGRF2-AS1. These candidates included S100-A9, ZN598, NRROS, ZMYM5, IF4A1, PDIP3, and PLCB4.

Conclusions

RASGRF2-AS1 is a novel key lncRNA involved in regulating HUVECs proliferation, apoptosis, and angiogenic ability. Consequently, RASGRF2-AS1 could be a promising target for treating arteriosclerosis obliterans.

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来源期刊

Gene Reports Biochemistry, Genetics and Molecular Biology-Genetics

CiteScore

3.30

自引率

7.70%

发文量

246

审稿时长

49 days

期刊介绍： Gene Reports publishes papers that focus on the regulation, expression, function and evolution of genes in all biological contexts, including all prokaryotic and eukaryotic organisms, as well as viruses. Gene Reports strives to be a very diverse journal and topics in all fields will be considered for publication. Although not limited to the following, some general topics include: DNA Organization, Replication & Evolution -Focus on genomic DNA (chromosomal organization, comparative genomics, DNA replication, DNA repair, mobile DNA, mitochondrial DNA, chloroplast DNA). Expression & Function - Focus on functional RNAs (microRNAs, tRNAs, rRNAs, mRNA splicing, alternative polyadenylation) Regulation - Focus on processes that mediate gene-read out (epigenetics, chromatin, histone code, transcription, translation, protein degradation). Cell Signaling - Focus on mechanisms that control information flow into the nucleus to control gene expression (kinase and phosphatase pathways controlled by extra-cellular ligands, Wnt, Notch, TGFbeta/BMPs, FGFs, IGFs etc.) Profiling of gene expression and genetic variation - Focus on high throughput approaches (e.g., DeepSeq, ChIP-Seq, Affymetrix microarrays, proteomics) that define gene regulatory circuitry, molecular pathways and protein/protein networks. Genetics - Focus on development in model organisms (e.g., mouse, frog, fruit fly, worm), human genetic variation, population genetics, as well as agricultural and veterinary genetics. Molecular Pathology & Regenerative Medicine - Focus on the deregulation of molecular processes in human diseases and mechanisms supporting regeneration of tissues through pluripotent or multipotent stem cells.