Deregulation of microRNA‑31a‑5p is involved in the development of primary hypertension by suppressing apoptosis of pulmonary artery smooth muscle cells via targeting TP53

Feng,Qiang; Tian,Tao; Liu,Junfeng; Zhang,Li; Qi,Jiangang; Lin,Xiaojuan

doi:10.3892/ijmm.2018.3597

Deregulation of microRNA‑31a‑5p is involved in the development of primary hypertension by suppressing apoptosis of pulmonary artery smooth muscle cells via targeting TP53

Authors:
- Qiang Feng
- Tao Tian
- Junfeng Liu
- Li Zhang
- Jiangang Qi
- Xiaojuan Lin
View Affiliations

Affiliations: Department of Laboratory, The People's Hospital of Tongchuan, Tongchuan, Shaanxi 727000, P.R. China, Department of Laboratory, Second Affiliated Hospital of Shaanxi Chinese Traditional Medicine, Xianyang, Shaanxi 712000, P.R. China, Department of Infection, The People's Hospital of Tongchuan, Tongchuan, Shaanxi 727000, P.R. China, Department of Gynecology and Obstetrics, The People's Hospital of Tongchuan, Tongchuan, Shaanxi 727000, P.R. China, Department of Laboratory, Tongchuan Hospital of Chinese Traditional Medicine, Tongchuan, Shaanxi 727000, P.R. China, Department of Cardiology, The People's Hospital of Tongchuan, Tongchuan, Shaanxi 727000, P.R. China
Published online on: March 29, 2018 https://doi.org/10.3892/ijmm.2018.3597
Pages: 290-298
Copyright: © Feng et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

The present study aimed to identify the association between microRNA (miRNA/miR)‑31a‑5p and the development of hypertension, and its potential molecular mechanism. Reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) and western blot analyses were performed to validate the candidate miRNA and genes involved in hypertension, following which an online miRNA database search, luciferase assay, and RT‑qPCR and western blot analyses were performed to confirm the interaction between miR‑31a‑5p and TP53. A MTT assay and flow cytometric analysis were utilized to determine the effect of miR‑31a‑5p on cell growth and apoptosis. The results revealed that miR‑31a‑5p and TP53 were the candidate miRNA and gene regulating hypertension, and that TP53 was the virtual target gene of miR‑31a‑5p with a binding site located in the TP53 3' untranslated region (3'UTR). It was confirmed by luciferase activity that miR‑31a‑5p markedly reduced the luciferase activity of the Luc‑wild‑type‑TP53‑3'UTR, whereas the mutated putative miR‑31a‑5p binding located on the TP53‑3'UTR was found to eliminate such an inhibitory effect. miR‑31a‑5p had no effect on specificity protein 1, E2F transcription factor 2 or forkhead box P3 luciferase activity. Smooth muscle cells collected from spontaneously hypertensive rats treated with gold nano‑particles containing anti‑rno‑miR‑31a‑5p exhibited a lower growth rate and a higher apoptotic rate. The results of the RT‑qPCR and western blot analyses showed that miR‑31a‑5p negatively regulated the expression of TP53, and transfection with the hsa‑miR‑31a‑5p mimic significantly promoted cell growth and inhibited cell apoptosis, whereas transfection with the anti‑hsa‑miR‑31a‑5p mimic significantly suppressed cell growth and induced cell apoptosis. Taken together, these findings indicated that miR‑31a‑5p is involved in hypertension via the accelerated proliferation of arterial smooth muscle cells and inhibition of apoptosis through targeting TP53.

View Figures

View References

1	Nwankwo T, Yoon SS, Burt V and Gu Q: Hypertension among adults in the United States: National Health and Nutrition Examination Survey, 2011–2012. NCHS Data Brief. 1–8. 2013.
2	Chiong M, Cartes-Saavedra B, Norambuena-Soto I, Mondaca-Ruff D, Morales PE, Garcia-Miguel M and Mellado R: Mitochondrial metabolism and the control of vascular smooth muscle cell proliferation. Front Cell Dev Biol. 2:722014. View Article : Google Scholar
3	Ragolia L, Palaia T, Paric E and Maesaka JK: Prostaglandin D2 synthase inhibits the exaggerated growth phenotype of spontaneously hypertensive rat vascular smooth muscle cells. J Biol Chem. 278:22175–22181. 2003. View Article : Google Scholar : PubMed/NCBI
4	Zheng GQ, Zhang GH, Xu XW, Wang JW, Yu LB, Zhang YN, Huang JW, Li YL, Braudt-Rauf PW and Xia ZL: Association of telomere length with chromosomal damage among Chinese workers exposed to Vinyl Chloride monomer. J Occup Environ Med. 59:e252–e256. 2017. View Article : Google Scholar : PubMed/NCBI
5	Pacurari M, Kafoury R, Tchounwou PB and Ndebele K: The Renin-Angiotensin-aldosterone system in vascular inflammation and remodeling. Int J Inflam. 2014:6893602014. View Article : Google Scholar : PubMed/NCBI
6	Mercer J, Figg N, Stoneman V, Braganza D and Bennett MR: Endogenous p53 protects vascular smooth muscle cells from apoptosis and reduces atherosclerosis in ApoE knockout mice. Circ Res. 96:667–674. 2005. View Article : Google Scholar : PubMed/NCBI
7	Giannakakou P, Sackett DL, Ward Y, Webster KR, Blagosklonny MV and Fojo T: p53 is associated with cellular microtubules and is transported to the nucleus by dynein. Nat Cell Biol. 2:709–717. 2000. View Article : Google Scholar : PubMed/NCBI
8	Krones-Herzig A, Adamson E and Mercola D: Early growth response 1 protein, an upstream gatekeeper of the p53 tumor suppressor, controls replicative senescence. Proc Natl Acad Sci USA. 100:3233–3238. 2003. View Article : Google Scholar : PubMed/NCBI
9	Mizuno S, Bogaard HJ, Kraskauskas D, Alhussaini A, Gomez-Arroyo J, Voelkel NF and Ishizaki T: p53 Gene deficiency promotes hypoxia-induced pulmonary hypertension and vascular remodeling in mice. Am J Physiol Lung Cell Mol Physiol. 300:L753–L761. 2011. View Article : Google Scholar : PubMed/NCBI
10	Sempere LF, Freemantle S, Pitha-Rowe I, Moss E, Dmitrovsky E and Ambros V: Expression profiling of mammalian microRNAs uncovers a subset of brain-expressed microRNAs with possible roles in murine and human neuronal differentiation. Genome Biol. 5:R132004. View Article : Google Scholar : PubMed/NCBI
11	Ambros V: The functions of animal microRNAs. Nature. 431:350–355. 2004. View Article : Google Scholar : PubMed/NCBI
12	Pan ZW, Lu YJ and Yang BF: MicroRNAs: A novel class of potential therapeutic targets for cardiovascular diseases. Acta Pharmacol Sin. 31:1–9. 2010. View Article : Google Scholar
13	Zapolska-Downar D, Siennicka A, Chelstowski K, Widecka K, Goracy I, Halasa M, Machalinski B and Naruszewicz M: Is there an association between angiotensin-converting enzyme gene polymorphism and functional activation of monocytes and macrophage in young patients with essential hypertension. J Hypertens. 24:1565–1573. 2006. View Article : Google Scholar : PubMed/NCBI
14	Kontaraki JE, Marketou ME, Parthenakis FI, Maragkoudakis S, Zacharis EA, Petousis S, Kochiadakis GE and Vardas PE: Hypertrophic and antihypertrophic microRNA levels in peripheral blood mononuclear cells and their relationship to left ventricular hypertrophy in patients with essential hypertension. J Am Soc Hypertens. 9:802–810. 2015. View Article : Google Scholar : PubMed/NCBI
15	Muller DN, Kvakan H and Luft FC: Immune-related effects in hypertension and target-organ damage. Curr Opin Nephrol Hypertens. 20:113–117. 2011. View Article : Google Scholar : PubMed/NCBI
16	Palao T, Sward K, Jongejan A, Moerland PD, de Vos J, van Weert A, Arribas SM, Groma G, vanBavel E and Bakker EN: Gene expression and microrna expression analysis in small arteries of spontaneously hypertensive rats. Evidence for ER stress. PLoS One. 10:e01370272015. View Article : Google Scholar
17	Jacquin S, Rincheval V, Mignotte B, Richard S, Humbert M, Mercier O, Londono-Vallejo A, Fadel E and Eddahibi S: Inactivation of p53 is sufficient to induce development of pulmonary hypertension in rats. PLoS One. 10:e01319402015. View Article : Google Scholar : PubMed/NCBI
18	Cai L, Li J, Zhang X, Lu Y, Wang J, Lyu X, Chen Y, Liu J, Cai H, Wang Y, et al: Gold nano-particles (AuNPs) carrying anti-EBV-miR-BART7-3p inhibit growth of EBV-positive nasopharyngeal carcinoma. Oncotarget. 6:7838–7850. 2015. View Article : Google Scholar : PubMed/NCBI
19	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
20	Kim BM and Choi MY: Non-canonical microRNAs miR-320 and miR-702 promote proliferation in Dgcr8-deficient embryonic stem cells. Biochem Biophys Res Commun. 426:183–189. 2012. View Article : Google Scholar : PubMed/NCBI
21	Dong Z, Zhong Z, Yang L, Wang S and Gong Z: MicroRNA-31 inhibits cisplatin-induced apoptosis in non-small cell lung cancer cells by regulating the drug transporter ABCB9. Cancer Lett. 343:249–257. 2014. View Article : Google Scholar
22	Joshi SR, Dhagia V, Gairhe S, Edwards JG, McMurtry IF and Gupte SA: MicroRNA-140 is elevated and mitofusin-1 is downregulated in the right ventricle of the Sugen5416/hypoxia/normoxia model of pulmonary arterial hypertension. Am J Physiol Heart Circ Physiol. 311:H689–H698. 2016. View Article : Google Scholar : PubMed/NCBI
23	Alzoubi A, Toba M, Abe K, O'Neill KD, Rocic P, Fagan KA, McMurtry IF and Oka M: Dehydroepiandrosterone restores right ventricular structure and function in rats with severe pulmonary arterial hypertension. Am J Physiol Heart Circ Physiol. 304:H1708–H1718. 2013. View Article : Google Scholar : PubMed/NCBI
24	Liu X, Cheng Y, Chen X, Yang J, Xu L and Zhang C: MicroRNA-31 regulated by the extracellular regulated kinase is involved in vascular smooth muscle cell growth via large tumor suppressor homolog 2. J Biol Chem. 286:42371–42380. 2011. View Article : Google Scholar : PubMed/NCBI
25	Pedrioli DM, Karpanen T, Dabouras V, Jurisic G, van de Hoek G, Shin JW, Marino D, Kalin RE, Leidel S, Cinelli P, et al: MiR-31 functions as a negative regulator of lymphatic vascular lineage-specific differentiation in vitro and vascular development in vivo. Mol Cell Biol. 30:3620–3634. 2010. View Article : Google Scholar : PubMed/NCBI
26	Shi B, Guo Y, Wang J and Gao W: Altered expression of microRNAs in the myocardium of rats with acute myocardial infarction. BMC Cardiovasc Disord. 10:112010. View Article : Google Scholar : PubMed/NCBI
27	Wang Y, Men M, Yang W, Zheng H and Xue S: MiR-31 downregulation protects against cardiac ischemia/reperfusion injury by targeting protein kinase C epsilon (PKCepsilon) directly. Cell Physiol Biochem. 36:179–190. 2015. View Article : Google Scholar
28	Huang CY, Kuo CH, Pai PY, Ho TJ, Lin YM, Chen RJ, Tsai FJ, Vijaya Padma V, Kuo WW and Huang CY: Inhibition of HSF2 SUMOylation via MEL18 upregulates IGF-IIR and leads to hypertension-induced cardiac hypertrophy. Int J Cardiol. S0167–5273:33565–33569. 2017.
29	Ueno T, Takagi H, Fukuda N, Takahashi A, Yao EH, Mitsumata M, Hiraoka-Yamamoto J, Ikeda K, Matsumoto K and Yamori Y: Cardiovascular remodeling and metabolic abnormalities in SHRSP.Z-Lepr(fa)/IzmDmcr rats as a new model of metabolic syndrome. Hypertens Res. 31:1021–1031. 2008. View Article : Google Scholar : PubMed/NCBI
30	Cannell IG, Merrick KA, Morandell S, Zhu CQ, Braun CJ, Grant RA, Cameron ER, Tsao MS, Hemann MT and Yaffe MB: A pleiotropic Rna-binding protein controls distinct cell cycle checkpoints to drive resistance of p53-defective tumors to chemotherapy. Cancer Cell. 28:623–637. 2015. View Article : Google Scholar : PubMed/NCBI
31	Muller P, Ceskova P and Vojtesek B: Hsp90 is essential for restoring cellular functions of temperature-sensitive p53 mutant protein but not for stabilization and activation of wild-type p53: Implications for cancer therapy. J Biol Chem. 280:6682–6691. 2005. View Article : Google Scholar
32	Zilfou JT and Lowe SW: Tumor suppressive functions of p53. Cold Spring Harb Perspect Biol. 1:a0018832009. View Article : Google Scholar :
33	Rodriguez-Campos A, Ruiz-Enriquez P, Faraudo S and Badimon L: Mitogen-induced p53 downregulation precedes vascular smooth muscle cell migration from healthy tunica media and proliferation. Arterioscler Thromb Vasc Biol. 21:214–219. 2001. View Article : Google Scholar : PubMed/NCBI
34	Vousden KH and Prives C: Blinded by the light: The growing complexity of p53. Cell. 137:413–431. 2009. View Article : Google Scholar : PubMed/NCBI
35	Laptenko O and Prives C: Transcriptional regulation by p53: One protein, many possibilities. Cell Death Differ. 13:951–961. 2006. View Article : Google Scholar : PubMed/NCBI
36	Tang X, Milyavsky M, Shats I, Erez N, Goldfinger N and Rotter V: Activated p53 suppresses the histone methyltransferase EZH2 gene. Oncogene. 23:5759–5769. 2004. View Article : Google Scholar : PubMed/NCBI
37	Carcagno AL, Marazita MC, Ogara MF, Ceruti JM, Sonzogni SV, Scassa ME, Giono LE and Canepa ET: E2F1-mediated upregulation of p19INK4d determines its periodic expression during cell cycle and regulates cellular proliferation. PLoS One. 6:e219382011. View Article : Google Scholar : PubMed/NCBI
38	Creighton CJ, Fountain MD, Yu Z, Nagaraja AK, Zhu H, Khan M, Olokpa E, Zariff A, Gunaratne PH, Matzuk MM, et al: Molecular profiling uncovers a p53-associated role for microRNA-31 in inhibiting the proliferation of serous ovarian carcinomas and other cancers. Cancer Res. 70:1906–1915. 2010. View Article : Google Scholar : PubMed/NCBI
39	Yang Y, Tarapore RS, Jarmel MH, Tetreault MP and Katz JP: p53 mutation alters the effect of the esophageal tumor suppressor KLF5 on keratinocyte proliferation. Cell Cycle. 11:4033–4039. 2012. View Article : Google Scholar : PubMed/NCBI
40	Ning Z, Zhu H, Li F, Liu Q, Liu G, Tan T, Zhang B, Chen S, Li G, Huang D, et al: Tumor suppression by miR-31 in esophageal carcinoma is p21-dependent. Genes Cancer. 5:436–444. 2014.