Long noncoding RNA <em>uc003pxg.1</em> regulates endothelial cell proliferation and migration via miR‑25‑5p in coronary artery disease

Li,Ping; Li,Ping; Li,Yuan; Li,Yuan; Chen,Lu; Chen,Lu; Ma,Xuexing; Ma,Xuexing; Yan,Xinxin; Yan,Xinxin; Yan,Meina; Yan,Meina; Qian,Buyun; Qian,Buyun; Wang,Feng; Wang,Feng; Xu,Jingyi; Xu,Jingyi; Yin,Juan; Yin,Juan; Xu,Guidong; Xu,Guidong; Sun,Kangyun; Sun,Kangyun

doi:10.3892/ijmm.2021.4993

Long noncoding RNA uc003pxg.1 regulates endothelial cell proliferation and migration via miR‑25‑5p in coronary artery disease

Authors:
- Ping Li
- Yuan Li
- Lu Chen
- Xuexing Ma
- Xinxin Yan
- Meina Yan
- Buyun Qian
- Feng Wang
- Jingyi Xu
- Juan Yin
- Guidong Xu
- Kangyun Sun
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Affiliations: Department of Central Laboratory, The Affiliated Suzhou Hospital of Nanjing Medical University, Suzhou Municipal Hospital, Gusu School, Nanjing Medical University, Suzhou, Jiangsu 215008, P.R. China, Department of Cardiology, The Affiliated Suzhou Hospital of Nanjing Medical University, Suzhou Municipal Hospital, Gusu School, Nanjing Medical University, Suzhou, Jiangsu 215008, P.R. China
Published online on: July 2, 2021 https://doi.org/10.3892/ijmm.2021.4993
Article Number: 160
Copyright: © Li et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Long noncoding RNAs (lncRNAs) have been reported to be associated with the progression of coronary artery disease (CAD). In our previous study, the levels of lncRNA uc003pxg.1 were upregulated in patients with CAD compared with those in control subjects. However, the role and underlying mechanism of the effects of uc003pxg.1 in CAD remain unknown. Therefore, the aim of the present study was to investigate the expression pattern and biological function of uc003pxg.1 in CAD. First, uc003pxg.1 expression levels were assessed in peripheral blood mononuclear cells isolated from patients with CAD by reverse transcription‑quantitative (RT‑q)PCR. The results demonstrated that the levels of uc003pxg.1 were significantly upregulated (~4.6‑fold) in samples from 80 patients with CAD compared with those in 80 healthy subjects. Subsequently, the present study demonstrated that small interfering RNA‑mediated uc003pxg.1 knockdown inhibited human umbilical vein endothelial cell (HUVEC) proliferation and migration, which was analyzed using the Cell Counting Kit‑8, cell cycle, EdU and Transwell assays. Additionally, the results of RT‑qPCR and western blot analyses revealed that uc003pxg.1 regulated the mRNA and protein levels of cyclin D1 and cyclin‑dependent kinase. Through high‑throughput sequencing and dual‑luciferase reporter assays, the present study demonstrated that microRNA (miR)‑25‑5p was a downstream target of uc003pxg.1. Further experiments verified that uc003pxg.1 regulated HUVEC proliferation and migration via miR‑25‑5p. The results of the present study may enhance the current understanding of the role of lncRNA uc003pxg.1 in CAD.

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1	Huang L, Zheng Y, Yuan X, Ma Y, Xie G, Wang W, Chen H and Shen L: Decreased frequencies and impaired functions of the CD31⁺ subpopulation in T_reg cells associated with decreased FoxP3 expression and enhanced T_reg cell defects in patients with coronary heart disease. Clin Exp Immunol. 187:441–454. 2017. View Article : Google Scholar
2	Jokinen E: Coronary artery disease in patients with congenital heart defects. J Intern Med. 288:383–389. 2020. View Article : Google Scholar : PubMed/NCBI
3	Malakar AK, Choudhury D, Halder B, Paul P, Uddin A and Chakraborty S: A review on coronary artery disease, its risk factors, and therapeutics. J Cell Physiol. 234:16812–16823. 2019. View Article : Google Scholar : PubMed/NCBI
4	Valgimigli M, Bueno H, Byrne RA, Collet JP, Costa F, Jeppsson A, Jüni P, Kastrati A, Kolh P, Mauri L, et al: 2017 ESC focused update on dual antiplatelet therapy in coronary artery disease developed in collaboration with EACTS: The Task Force for dual antiplatelet therapy in coronary artery disease of the European Society of Cardiology (ESC) and of the European association for cardio-thoracic surgery (EACTS). Eur Heart J. 39:213–260. 2018. View Article : Google Scholar
5	Cai Y, Yang Y, Chen X, Wu G, Zhang X, Liu Y, Yu J, Wang X, Fu J, Li C, et al: Circulating 'lncRNA OTTHUMT00000387022' from monocytes as a novel biomarker for coronary artery disease. Cardiovasc Res. 112:714–724. 2016. View Article : Google Scholar : PubMed/NCBI
6	Huang Y: The novel regulatory role of lncRNA-miRNA-mRNA axis in cardiovascular diseases. J Cell Mol Med. 22:5768–5775. 2018. View Article : Google Scholar : PubMed/NCBI
7	Bhan A and Mandal SS: LncRNA HOTAIR: A master regulator of chromatin dynamics and cancer. Biochim Biophys Acta. 1856:151–164. 2015.PubMed/NCBI
8	Derrien T, Johnson R, Bussotti G, Tanzer A, Djebali S, Tilgner H, Guernec G, Martin D, Merkel A, Knowles DG, et al: The GENCODE v7 catalog of human long noncoding RNAs: Analysis of their gene structure, evolution, and expression. Genome Res. 22:1775–1789. 2012. View Article : Google Scholar : PubMed/NCBI
9	Sanfilippo PG and Hewitt AW: Translating the ENCyclopedia Of DNA elements project findings to the clinic: ENCODE's implications for eye disease. Clin Exp Ophthalmol. 42:78–83. 2014. View Article : Google Scholar : PubMed/NCBI
10	Bhan A, Soleimani M and Mandal SS: Long noncoding RNA and cancer: A new paradigm. Cancer Res. 77:3965–3981. 2017. View Article : Google Scholar : PubMed/NCBI
11	Kaikkonen MU, Lam MT and Glass CK: Non-coding RNAs as regulators of gene expression and epigenetics. Cardiovasc Res. 90:430–440. 2011. View Article : Google Scholar : PubMed/NCBI
12	Greco S, Gorospe M and Martelli F: Noncoding RNA in age-related cardiovascular diseases. J Mol Cell Cardiol. 83:142–155. 2015. View Article : Google Scholar : PubMed/NCBI
13	Li L, Wang L, Li H, Han X, Chen S, Yang B, Hu Z, Zhu H, Cai C, Chen J, et al: Characterization of LncRNA expression profile and identification of novel LncRNA biomarkers to diagnose coronary artery disease. Atherosclerosis. 275:359–367. 2018. View Article : Google Scholar : PubMed/NCBI
14	Zhang H, Ji N, Gong X, Ni S and Wang Y: NEAT1/miR-140-3p/MAPK1 mediates the viability and survival of coronary endothelial cells and affects coronary atherosclerotic heart disease. Acta Biochim Biophys Sin (Shanghai). 52:967–974. 2020. View Article : Google Scholar
15	Zhang Z, Gao W, Long QQ, Zhang J, Li YF, Liu DC, Yan JJ, Yang ZJ and Wang LS: Increased plasma levels of lncRNA H19 and LIPCAR are associated with increased risk of coronary artery disease in a Chinese population. Sci Rep. 7:74912017. View Article : Google Scholar : PubMed/NCBI
16	Tang Y, Jin X, Xiang Y, Chen Y, Shen CX, Zhang YC and Li YG: The lncRNA MALAT1 protects the endothelium against ox-LDL-induced dysfunction via upregulating the expression of the miR-22-3p target genes CXCR2 and AKT. FEBS Lett. 589:3189–3196. 2015. View Article : Google Scholar : PubMed/NCBI
17	Michalik KM, You X, Manavski Y, Doddaballapur A, Zörnig M, Braun T, John D, Ponomareva Y, Chen W, Uchida S, et al: Long noncoding RNA MALAT1 regulates endothelial cell function and vessel growth. Circ Res. 114:1389–1397. 2014. View Article : Google Scholar : PubMed/NCBI
18	Motterle A, Pu X, Wood H, Xiao Q, Gor S, Ng FL, Chan K, Cross F, Shohreh B, Poston RN, et al: Functional analyses of coronary artery disease associated variation on chromosome 9p21 in vascular smooth muscle cells. Hum Mol Genet. 21:4021–4029. 2012. View Article : Google Scholar : PubMed/NCBI
19	Ivanov D, Philippova M, Allenspach R, Erne P and Resink T: T-cadherin upregulation correlates with cell-cycle progression and promotes proliferation of vascular cells. Cardiovasc Res. 64:132–143. 2004. View Article : Google Scholar : PubMed/NCBI
20	Wang G, Li Y, Peng Y, Tang J and Li H: Association of polymorphisms in MALAT1 with risk of coronary atherosclerotic heart disease in a Chinese population. Lipids Health Dis. 17:752018. View Article : Google Scholar : PubMed/NCBI
21	Wu Z, He Y, Li D, Fang X, Shang T, Zhang H and Zheng X: Long noncoding RNA MEG3 suppressed endothelial cell proliferation and migration through regulating miR-21. Am J Transl Res. 9:3326–3335. 2017.PubMed/NCBI
22	Li P, Yan X, Xu G, Pang Z, Weng J, Yin J, Li M, Yu L, Chen Q and Sun K: A novel plasma lncRNA ENST00000416361 is upregulated in coronary artery disease and is related to inflammation and lipid metabolism. Mol Med Rep. 21:2375–2384. 2020.PubMed/NCBI
23	Wenger NK: 2011 ACCF/AHA focused update of the guidelines for the management of patients with unstable angina/Non-ST-elevation myocardial infarction (updating the 2007 guideline): Highlights for the clinician. Clin Cardiol. 35:3–8. 2012. View Article : Google Scholar
24	Jia Y, Xu H, Li Y, Wei C, Guo R, Wang F, Wu Y, Liu J, Jia J, Yan J, et al: A modified ficoll-paque gradient method for isolating mononuclear cells from the peripheral and umbilical cord blood of humans for biobanks and clinical laboratories. Biopreserv Biobank. 16:82–91. 2018. View Article : Google Scholar
25	Trapnell C, Hendrickson DG, Sauvageau M, Goff L, Rinn JL and Pachter L: Differential analysis of gene regulation at transcript resolution with RNA-seq. Nat Biotechnol. 31:46–53. 2013. View Article : Google Scholar
26	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar
27	Wang L, Feng Z, Wang X, Wang X and Zhang X: DEGseq: An R package for identifying differentially expressed genes from RNA-seq data. Bioinformatics. 26:136–138. 2010. View Article : Google Scholar
28	Liu H, Lei C, He Q, Pan Z, Xiao D and Tao Y: Nuclear functions of mammalian MicroRNAs in gene regulation, immunity and cancer. Mol Cancer. 17:642018. View Article : Google Scholar : PubMed/NCBI
29	Wang Y, Tao B, Li J, Mao X, He W and Chen Q: Melatonin inhibits the progression of oral squamous cell carcinoma via inducing miR-25-5p expression by directly targeting NEDD9. Front Oncol. 10:5435912020. View Article : Google Scholar :
30	Zhang Q, Liu C, Li Q, Li J, Wu Y and Liu J: MicroRNA-25-5p counteracts oxidized LDL-induced pathological changes by targeting neuronal growth regulator 1 (NEGR1) in human brain micro-vessel endothelial cells. Biochimie. 165:141–149. 2019. View Article : Google Scholar : PubMed/NCBI
31	Niccoli G, Montone RA, Sabato V and Crea F: Role of allergic inflammatory cells in coronary artery disease. Circulation. 138:1736–1748. 2018. View Article : Google Scholar : PubMed/NCBI
32	Weber C and Noels H: Atherosclerosis: Current pathogenesis and therapeutic options. Nat Med. 17:1410–1422. 2011. View Article : Google Scholar : PubMed/NCBI
33	Knorr M, Münzel T and Wenzel P: Interplay of NK cells and monocytes in vascular inflammation and myocardial infarction. Front Physiol. 5:2952014. View Article : Google Scholar : PubMed/NCBI
34	Jaipersad AS, Shantsila A, Lip GY and Shantsila E: Expression of monocyte subsets and angiogenic markers in relation to carotid plaque neovascularization in patients with pre-existing coronary artery disease and carotid stenosis. Ann Med. 46:530–538. 2014. View Article : Google Scholar : PubMed/NCBI
35	Yang T, Chen T, Li Y, Gao L, Zhang S, Wang T and Chen M: Downregulation of miR-25 modulates non-small cell lung cancer cells by targeting CDC42. Tumour Biol. 36:1903–1911. 2015. View Article : Google Scholar
36	Wang C, Wang X, Su Z, Fei H, Liu X and Pan Q: MiR-25 promotes hepatocellular carcinoma cell growth, migration and invasion by inhibiting RhoGDI1. Oncotarget. 6:36231–36244. 2015. View Article : Google Scholar : PubMed/NCBI
37	Jung JH, Shin EA, Kim JH, Sim DY, Lee H, Park JE, Lee HJ and Kim SH: NEDD9 inhibition by miR-25-5p activation is critically involved in co-treatment of melatonin- and pterostilbene-induced apoptosis in colorectal cancer cells. Cancers (Basel). 11:16842019. View Article : Google Scholar
38	Yao Y, Song T, Xiong G, Wu Z, Li Q, Xia H and Jiang X: Combination of peripheral blood mononuclear cell miR-19b-5p miR-221, miR-25-5p and hypertension correlates with an increased heart failure risk in coronary heart disease patients. Anatol J Cardiol. 20:100–109. 2018.PubMed/NCBI
39	Wu JB, Ye XH, Xian SX and Dong MG: Expressions of SERCA2a and miR-25-3p/5p in myocardium of rats with heart failure and therapeutic effects of Xiefei Lishui recipe. Zhongguo Ying Yong Sheng Li Xue Za Zhi. 33:146–150. 2017.In Chinese. PubMed/NCBI
40	Meseure D, Drak Alsibai K, Nicolas A, Bieche I and Morillon A: Long noncoding RNAs as new architects in cancer epigenetics, prognostic biomarkers, and potential therapeutic targets. Biomed Res Int. 2015:3202142015. View Article : Google Scholar : PubMed/NCBI
41	Rizzacasa B, Amati F, Romeo F, Novelli G and Mehta JL: Epigenetic modification in coronary atherosclerosis: JACC review topic of the week. J Am Coll Cardiol. 74:1352–1365. 2019. View Article : Google Scholar : PubMed/NCBI
42	Zhang YH, Pan X, Zeng T, Chen L, Huang T and Cai YD: Identifying the RNA signatures of coronary artery disease from combined lncRNA and mRNA expression profiles. Genomics. 112:4945–4958. 2020. View Article : Google Scholar : PubMed/NCBI