Aberrant DNA methylation of the PDGF gene in homocysteine‑mediated VSMC proliferation and its underlying mechanism

Han,Xue‑Bo; Zhang,Hui‑Ping; Cao,Cheng‑Jian; Wang,Yan‑Hua; Tian,Jue; Yang,Xiao‑Ling; Yang,An‑Ning; Wang,Jie; Jiang,Yi‑Deng; Xu,Hua

doi:10.3892/mmr.2014.2249

Aberrant DNA methylation of the PDGF gene in homocysteine‑mediated VSMC proliferation and its underlying mechanism

Authors:
- Xue‑Bo Han
- Hui‑Ping Zhang
- Cheng‑Jian Cao
- Yan‑Hua Wang
- Jue Tian
- Xiao‑Ling Yang
- An‑Ning Yang
- Jie Wang
- Yi‑Deng Jiang
- Hua Xu
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Affiliations: Department of Laboratory Medicine, Ningxia Medical University, Yinchuan, Ningxia Hui Autonomous Region 750004, P.R. China, Department of Prenatal Diagnosis Center, General Hospital of Ningxia Medical University, Yinchuan, Ningxia Hui Autonomous Region 750004, P.R. China, Department of Pathophysiology, Basic Medical School, Ningxia Medical University, Yinchuan, Ningxia Hui Autonomous Region 750004, P.R. China, Center of Scientific Technology, Ningxia Medical University, Yinchuan, Ningxia Hui Autonomous Region 750004, P.R. China
Published online on: May 20, 2014 https://doi.org/10.3892/mmr.2014.2249
Pages: 947-954

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Abstract

It is well established that homocysteine (Hcy) is an independent risk factor for atherosclerosis (AS), which is characterized by vascular smooth muscle cell (VSMC) proliferation. However, the molecular mechanism underlying AS in VSMCs is yet to be elucidated. The aim of this study was to investigate the potential involvement of aberrant DNA methylation of the platelet‑derived growth factor (PDGF) gene in Hcy‑mediated VSMC proliferation and its underlying mechanism. Cultured human VSMCs were treated with varying concentrations of Hcy. VSMC proliferation, PDGF mRNA and protein expression and PDGF promoter demethylation showed a dose‑dependent increase with Hcy concentration, suggesting an association among them. Cell cycle analysis revealed a decreased proportion of VSMCs in G0/G1 and an increased proportion in S phase, indicating that VSMC proliferation was increased under Hcy treatment. Furthermore, S‑adenosylhomocysteine (SAH) levels were observed to increase and those of S‑adenosylmethionine (SAM) were observed to decrease. The consequent decrease in the ratio of SAM/SAH may partially explain the hypomethylation of PDGF with Hcy treatment. Folate treatment exhibited an antagonistic effect against Hcy‑induced VSMC proliferation, aberrant PDGF methylation and PDGF expression. These data suggest that Hcy may stimulate VSMC proliferation through the PDGF signaling pathway by affecting the epigenetic regulation of PDGF through the demethylation of its promoter region. These findings may provide novel insight into the molecular association between aberrant PDGF gene demethylation and the proliferation of VSMCs in Hcy‑associated AS.

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1	Zhang J, Guo C, Wang R, Huang L, Liang W, Liu R and Sun B: An Egr-1-specific DNAzyme regulates Egr-1 and proliferating cell nuclear antigen expression in rat vascular smooth muscle cells. Exp Ther Med. 5:1371–1374. 2013.PubMed/NCBI
2	Little PJ, Rostam MA, Piva TJ, et al: Suramin inhibits PDGF-stimulated receptor phosphorylation, proteoglycan synthesis and glycosaminoglycan hyperelongation in human vascular smooth muscle cells. J Pharm Pharmacol. 65:1055–1063. 2013. View Article : Google Scholar
3	Jia G, Cheng G, Gangahar DM and Agrawal DK: Involvement of connexin 43 in angiotensin II-induced migration and proliferation of saphenous vein smooth muscle cells via the MAPK-AP-1 signaling pathway. J Mol Cell Cardiol. 44:882–890. 2008. View Article : Google Scholar : PubMed/NCBI
4	Adhikari N, Basi DL, Townsend D, Rusch M, Mariash A, Mullegama S, Watson A, Larson J, Tan S, Lerman B, Esko JD, Selleck SB and Hall JL: Heparan sulfate Ndst1 regulates vascular smooth muscle cell proliferation, vessel size and vascular remodeling. J Mol Cell Cardiol. 49:287–293. 2010. View Article : Google Scholar : PubMed/NCBI
5	Luo X, Xiao Y, Song F, Yang Y, Xia M and Ling W: Increased plasma S-adenosyl-homocysteine levels induce the proliferation and migration of VSMCs through an oxidative stress-ERK1/2 pathway in apoE(−/−) mice. Cardiovasc Res. 95:241–250. 2012.PubMed/NCBI
6	Zhang D, Chen Y, Xie X, Liu J, Wang Q, Kong W and Zhu Y: Homocysteine activates vascular smooth muscle cells by DNA demethylation of platelet-derived growth factor in endothelial cells. J Mol Cell Cardiol. 53:487–496. 2012. View Article : Google Scholar : PubMed/NCBI
7	Janda K, Aksamit D, Drozdz M, et al: Influence of elevated homocystein level and selected lipid parameters in kidney transplant patients on the progression of atherosclerotic changes assessed by intima-media thickness index (CCA-IMT). Przegl Lek. 69:670–674. 2012.(In Polish).
8	Huidobro C, Fernandez AF and Fraga MF: The role of genetics in the establishment and maintenance of the epigenome. Cell Mol Life Sci. 70:1543–1573. 2013. View Article : Google Scholar : PubMed/NCBI
9	Tisdale MJ: Potentiation of the growth inhibitory effects of adenosine 3′,5′-monophosphate analogues by homocysteine. Biochem Pharmacol. 31:979–982. 1982.
10	Zhang CY, Wang NN, Zhang YH, Feng QZ, Yang CW and Liu B: DNA methylation involved in proline accumulation in response to osmotic stress in rice (Oryza sativa). Genet Mol Res. 12:1269–1277. 2013. View Article : Google Scholar : PubMed/NCBI
11	He Y, Cui Y, Wang W, Gu J, Guo S, Ma K and Luo X: Hypomethylation of the hsa-miR-191 locus causes high expression of hsa-mir-191 and promotes the epithelial-to-mesenchymal transition in hepatocellular carcinoma. Neoplasia. 13:841–853. 2011.PubMed/NCBI
12	Amatruda JF, Ross JA, Christensen B, et al: DNA methylation analysis reveals distinct methylation signatures in pediatric germ cell tumors. BMC Cancer. 13:3132013. View Article : Google Scholar : PubMed/NCBI
13	Yideng J, Zhihong L, Jiantuan X, Jun C, Guizhong L and Shuren W: Homocysteine-mediated PPARalpha, gamma DNA methylation and its potential pathogenic mechanism in monocytes. DNA Cell Biol. 27:143–150. 2008. View Article : Google Scholar : PubMed/NCBI
14	Sahni A, Wang N and Alexis J: UAP56 is an important mediator of angiotensin II/platelet derived growth factor induced vascular smooth muscle cell DNA synthesis and proliferation. Biochem Biophys Res Commun. 431:636–640. 2013. View Article : Google Scholar : PubMed/NCBI
15	Thevathasan JV, Tan E, Zheng H, Lin YC, Li Y, Inoue T and Fivaz M: The small GTPase HRas shapes local PI3K signals through positive feedback and regulates persistent membrane extension in migrating fibroblasts. Mol Biol Cell. 24:2228–2237. 2013. View Article : Google Scholar
16	Chandra A and Angle N: VEGF inhibits PDGF-stimulated calcium signaling independent of phospholipase C and protein kinase C. J Surg Res. 131:302–309. 2006. View Article : Google Scholar : PubMed/NCBI
17	Kojima N, Hori M, Murata T, Morizane Y and Ozaki H: Different profiles of Ca²⁺ responses to endothelin-1 and PDGF in liver myofibroblasts during the process of cell differentiation. Br J Pharmacol. 151:816–827. 2007.
18	Hattori N, Abe T, Hattori N, et al: Preference of DNA methyltransferases for CpG islands in mouse embryonic stem cells. Genome Res. 14:1733–1740. 2004. View Article : Google Scholar : PubMed/NCBI
19	Dai Y, Mercanti F, Dai D, Wang X, Ding Z, Pothineni NV and Mehta JL: LOX-1, a bridge between GLP-1R and mitochondrial ROS generation in human vascular smooth muscle cells. Biochem Biophys Res Commun. 437:62–66. 2013. View Article : Google Scholar : PubMed/NCBI
20	Whitman SC: A practical approach to using mice in atherosclerosis research. Clin Biochem Rev. 25:81–93. 2004.PubMed/NCBI
21	Shi ZD and Tarbell JM: Fluid flow mechanotransduction in vascular smooth muscle cells and fibroblasts. Ann Biomed Eng. 39:1608–1619. 2011. View Article : Google Scholar : PubMed/NCBI
22	Xie C, Ritchie RP, Huang H, Zhang J and Chen YE: Smooth muscle cell differentiation in vitro: models and underlying molecular mechanisms. Arterioscler Thromb Vasc Biol. 31:1485–1494. 2011. View Article : Google Scholar : PubMed/NCBI
23	Yideng J, Jianzhong Z, Ying H, Juan S, Jinge Z, Shenglan W, Xiaoqun H and Shuren W: Homocysteine-mediated expression of SAHH, DNMTs, MBD2, and DNA hypomethylation potential pathogenic mechanism in VSMCs. DNA Cell Biol. 26:603–611. 2007. View Article : Google Scholar : PubMed/NCBI
24	Liu X, Shen J, Zhan R, et al: Proteomic analysis of homocysteine induced proliferation of cultured neonatal rat vascular smooth muscle cells. Biochim Biophys Acta. 1794:177–184. 2009. View Article : Google Scholar : PubMed/NCBI
25	Nazarenko I, Hede SM, He X, Hedrén A, Thompson J, Lindström MS and Nistér M: PDGF and PDGF receptors in glioma. Ups J Med Sci. 117:99–112. 2012. View Article : Google Scholar : PubMed/NCBI
26	Sakata Y, Xiang F, Chen Z, Kiriyama Y, Kamei CN, Simon DI and Chin MT: Transcription factor CHF1/Hey2 regulates neointimal formation in vivo and vascular smooth muscle proliferation and migration in vitro. Arterioscler Thromb Vasc Biol. 24:2069–2074. 2004. View Article : Google Scholar : PubMed/NCBI
27	Zhang H, Jia X, Han F, Zhao J, Zhao Y, Fan Y and Yuan X: Dual-delivery of VEGF and PDGF by double-layered electrospun membranes for blood vessel regeneration. Biomaterials. 34:2202–2212. 2013. View Article : Google Scholar : PubMed/NCBI
28	Li JC, Pan JQ, Huang GQ, Tan X, Sun WD, Liu YJ and Wang XL: Expression of PDGF-beta receptor in broilers with pulmonary hypertension induced by cold temperature and its association with pulmonary vascular remodeling. Res Vet Sci. 88:116–121. 2010. View Article : Google Scholar : PubMed/NCBI
29	Takimoto T, Suzuki K, Arisaka H, Murata T, Ozaki H and Koyama N: Effect of N-(p-coumaroyl)serotonin and N-feruloylserotonin, major anti-atherogenic polyphenols in safflower seed, on vasodilation, proliferation and migration of vascular smooth muscle cells. Mol Nutr Food Res. 55:1561–1571. 2011. View Article : Google Scholar
30	Choi BK, Cha BY, Yagyu T, Woo JT and Ojika M: Sponge-derived acetylenic alcohols, petrosiols, inhibit proliferation and migration of platelet-derived growth factor (PDGF)-induced vascular smooth muscle cells. Bioorg Med Chem. 21:1804–1810. 2013. View Article : Google Scholar
31	Sirois MG, Simons M and Edelman ER: Antisense oligonucleotide inhibition of PDGFR-beta receptor subunit expression directs suppression of intimal thickening. Circulation. 95:669–676. 1997. View Article : Google Scholar
32	Englesbe MJ, Hawkins SM, Hsieh PC, Daum G, Kenagy RD and Clowes AW: Concomitant blockade of platelet-derived growth factor receptors alpha and beta induces intimal atrophy in baboon PTFE grafts. J Vasc Surg. 39:440–446. 2004. View Article : Google Scholar : PubMed/NCBI
33	Kwon HJ, Kim GE, Lee YT, Jeong MS, Kang I, Yang D and Yeo EJ: Inhibition of platelet-derived growth factor receptor tyrosine kinase and downstream signaling pathways by Compound C. Cell Signal. 25:883–897. 2013. View Article : Google Scholar : PubMed/NCBI
34	Chen NC, Yang F, Capecci LM, et al: Regulation of homocysteine metabolism and methylation in human and mouse tissues. FASEB J. 24:2804–2817. 2010. View Article : Google Scholar : PubMed/NCBI
35	Hiltunen MO, Turunen MP, Häkkinen TP, et al: DNA hypomethylation and methyltransferase expression in atherosclerotic lesions. Vasc Med. 7:5–11. 2002. View Article : Google Scholar : PubMed/NCBI
36	Duncan TM, Reed MC and Nijhout HF: The relationship between intracellular and plasma levels of folate and metabolites in the methionine cycle: a model. Mol Nutr Food Res. 57:628–636. 2013. View Article : Google Scholar : PubMed/NCBI