Linc00152 Sponges Mir-4767 To Regulating Function Of Endothelial Cells

BackgroundIncreasing evidence reveals that IncRNAs participate in nearly all aspects of gene expression regulation,including epigenetics,transcription and post-transcription,and are involved in many biological processes under both physiological and pathological conditions,such as cell growth and movement,differentiation and reprogramming,and stress response.Fine regulation of gene expression,at the transcriptional and post-transcriptional levels,is required in the response of mammalian cells to internal and external stimuli.Transcriptional regulation achieves by alternation in the composition of transcription factor-promoter complex,while post-transcriptional regulation is uausally mediated by alternative splicing of RNA transcripts,alterations in the stability and transport of mRNAs.LncRNAs are the most prevalent and functionally diverse regulator in gene fine regulation.They can function at relatively low expression level and their regulation can be triggered by stimuli during various pathological processes including vascular injury and remodeling.Vascular endothelial cells(VECs)lie in the innermost of blood vessels.They are the most abundant component of blood vessel,and they are vulnerable to stress.Dysfunction of VECs is important in pathogenesis of many cardiovascular diseases,such as arteriosclerosis,arteritis,thrombus and plaque erosion.Protein-coding genes involved in the regulaion of VEC functions have been relatively well studied.The role of lncRNAs in regulating atherosclerosis and other cardiovascular diseases has attracted interest,but the exact role of lncRNAs in VEC functions was largely unknown.Linc00152 has been shown to upregulated in several carcinoma tissues,promote cell proliferation and metastasis,and inhibit cell apoptosis.However,its role in non-cancer biological processes was unknown.Recent studies revealed that linc00152 could be expressed in the circulatory system and that it responds to chemical stresses,suggesting that its function might be not restricted in carcinogenesis.ObjectiveIn order to explore the role of a intergenic IncRNA named linc00152 in the apoptosis and migration of VECs.MethodHuman umbilical vein endothelial cells(HUVECs)were cultured in Dulbecco's Modified Eagle Medium(DMEM,Invitrogen,Carlsbad,CA,USA)supplemented with 10%fetal bovine serum(FBS,Invitrogen),100U/mL penicillin(Invitrogen)and 100 mg/mL streptomycin(Invitrogen).The cells were incubated at 37? in a humidified and 5%CO2 atmosphere.For treatment,HUVECs were subcultured in 12-well culture plates at a density of 105/well.On reaching 50%confluence,ox-LDL(dissolved in PBS containing 0.08 ?M EDTA-Na2)was added into the cells at the final concentration of 50?g/mL,100?g/mL,150?g/mL and 200 ?g/mL.As a control,an equal volume of PBS containing 0.08?M EDTA-Na2 was incubated with the cells.The ex-pression of lincOO 152 mRNA in cells was detected by transfection of vectors,quantitative reverse transcription-PCR(qRT-PCR),Western blotting,Luciferase constructs and activity detection method.Values were expressed as means ± SEM.Statistics were calculated with SPSS statistics 23.0 software.Multiple comparisons were assessed by one-way ANOVA followed by Dunnett's tests.The difference between groups was considered statistically significant if P<0.05.ResultWe found that linc00152 was downregulated in human umbilical vein VECs(HUVECs)in a dose-dependent and time-dependent manner following treatment with oxLDL,which is a typical proinflammatory factor in the initiation and progression of vascular endothelial dysfunction.Gain-and loss-function experiments indicated that linc00152 distinctly inhibited apoptosis and improved migration in oxLDL-treated HUVECs.By sponging miR-4767,linc00152 positively regulated the expression of Bc12L12 and EGFR proteins.Moreover,blocking miR-4767 rescued the decrease of Bc12L12 and EGFR caused by linc00152 knockdown,as well as the changes in cell apoptosis and migration.ConclusionOur findings propose a novel role of linc00152 in the improvement of vascular endothelial function and a potential target for the therapy of some cardiovascular diseases.