Tieling LI, Xiaochun LU, Shi-Tang MA, Bo ZHOU

MiR-9 promotes the phenotypic switch of vascular smooth muscle cells by targeting KLF5

Background/aim: Diabetic vascular smooth muscle cells (VSMCs) are characterized by increased proliferation and migration. Smallnoncoding microRNAs (miRNAs) have been considered critical modulators of the VSMC phenotypic switch after an environmentalstimulus. However, microRNA in high glucose-induced proinflammation and its atherogenic effect is still ambiguous.Materials and methods: The technique of qRT-PCR was used to examine the expression of miR-9 in VSMCs. The downstream signalingprotein relative to miR-9 regulation, Krüppel-like factor 5, and some marker genes of contractile VSMCs were analyzed by westernblotting and qRT-PCR. Luciferase reporter assay was used to detect the expression of KLF5, which is regulated by miR-9. To examinethe function of a miR-9 inhibitor in VSMC proliferation and migration, VSMC proliferation and migration assays were performed.Results: Reduced transcriptional levels of miR-9 and expression of specific genes of contractile VSMCs were observed in the SMC cellline C-12511 treated with high glucose and SMCs, which were isolated from db/db mice. Moreover, the activity of KLF5 3′-UTR wasdramatically reduced by a miR-9 mimic and increased by a miR-9 inhibitor. The proliferation and migration of SMCs were reduced bythe miR-9 mimic.Conclusion: miR-9 inhibits the proliferation and migration of SMC by targeting KLF5 in db/db mice.

PDF

___

1. Lachin JM, Orchard TJ, Nathan DM, Group DER. Update on cardiovascular outcomes at 30 years of the diabetes control and complications trial/epidemiology of diabetes interventions and complications study. Diabetes Care 2014; 37: 39-43.
2. Johnson JL. Emerging regulators of vascular smooth muscle cell function in the development and progression of atherosclerosis. Cardiovasc Res 2014; 103: 452-460.
3. Bennett MR, Sinha S, Owens GK. Vascular smooth muscle cells in atherosclerosis. Circ Res 2016; 118: 692-702.
4. Li T, Yang GM, Zhu Y, Wu Y, Chen XY, Lan D, Tian KL, Liu LM. Diabetes and hyperlipidemia induce dysfunction of VSMCs: contribution of the metabolic inflammation/miRNA pathway. Am J Physiol Endocrinol Metab 2015; 308: E257-E269.
5. Verspohl EJ, Hagemann J, Lempka M. Vascular smooth muscle cells (VSMC) proliferation of streptozotocin-diabetic animals induced by diadenosine polyphosphates. Exp Clin Endocrinol Diabetes 2004; 112: 142-147.
6. Zhang MJ, Zhou Y, Chen L, Wang YQ, Wang X, Pi Y, Gao CY, Li JC, Zhang LL. An overview of potential molecular mechanisms involved in VSMC phenotypic modulation. Histochem Cell Biol 2016; 145: 119-130.
7. Farrugia MK, Vanderbilt DB, Salkeni MA, Ruppert JM. Kruppel-like pluripotency factors as modulators of cancer cell therapeutic responses. Cancer Res 2016; 76: 1677-1682.
8. Ghaleb AM, Nandan MO, Chanchevalap S, Dalton WB, Hisamuddin IM, Yang VW. Kruppel-like factors 4 and 5: the yin and yang regulators of cellular proliferation. Cell Res 2005; 15: 92-96.
9. Cheng Y, Liu X, Yang J, Lin Y, Xu DZ, Lu Q, Deitch EA, Huo Y, Delphin ES, Zhang C. MicroRNA-145, a novel smooth muscle cell phenotypic marker and modulator, controls vascular neointimal lesion formation. Circ Res 2009; 105: 158-166.
10. Ha JM, Yun SJ, Jin SY, Lee HS, Kim SJ, Shin HK, Bae SS. Regulation of vascular smooth muscle phenotype by crossregulation of Kruppel-like factors. Korean J Physiol Pharmacol 2017; 21: 37-44.
11. Liu JY, Lu JB, Xu Y. MicroRNA-153 inhibits the proliferation and invasion of human laryngeal squamous cell carcinoma by targeting KLF5. Exp Ther Med 2016; 11: 2503-2508.
12. Li P, Zhu N, Yi B, Wang N, Chen M, You X, Zhao X, Solomides CC, Qin Y, Sun J. MicroRNA-663 regulates human vascular smooth muscle cell phenotypic switch and vascular neointimal formation. Circ Res 2013; 113: 1117-1127.
13. Tan J, Yang L, Liu C, Yan Z. MicroRNA-26a targets MAPK6 to inhibit smooth muscle cell proliferation and vein graft neointimal hyperplasia. Sci Rep 2017; 7: 46602.
14. Huang SC, Wang M, Wu WB, Wang R, Cui J, Li W, Li ZL, Li W, Wang SM. Mir-22-3p inhibits arterial smooth muscle cell proliferation and migration and neointimal hyperplasia by targeting HMGB1 in arteriosclerosis obliterans. Cell Physiol Biochem 2017; 42: 2492-2506.
15. Ham O, Lee SY, Song BW, Lee CY, Lee J, Seo HH, Kim SW, Lim S, Kim IK, Lee S, Hwang KC. Small molecule-mediated induction of miR-9 suppressed vascular smooth muscle cell proliferation and neointima formation after balloon injury. Oncotarget 2017; 8: 93360-93372.
16. Shan F, Li J, Huang QY. HIF-1 alpha-induced up-regulation of miR-9 contributes to phenotypic modulation in pulmonary artery smooth muscle cells during hypoxia. J Cell Physiol 2014; 229: 1511-1520.
17. Yao H, Ma R, Yang L, Hu G, Chen X, Duan M, Kook Y, Niu F, Liao K, Fu M, Hu G, Kolattukudy P, Buch S. MiR-9 promotes microglial activation by targeting MCPIP1. Nat Commun 2014; 5: 4386.
18. Reddy MA, Das S, Zhuo C, Jin W, Wang M, Lanting L, Natarajan R. Regulation of vascular smooth muscle cell dysfunction under diabetic conditions by miR-504. Arterioscler Thromb Vasc Biol 2016; 36: 864-873.
19. Niu C, Wang X, Zhao M, Cai T, Liu P, Li J, Willard B, Zu L, Zhou E, Li Y, Pan B, Yang F, Zheng L. Macrophage foam cellderived extracellular vesicles promote vascular smooth muscle cell migration and adhesion. J Am Heart Assoc 2016; 5.
20. Coolen M, Katz S, Bally-Cuif L. miR-9: a versatile regulator of neurogenesis. Front Cell Neurosci 2013; 7: 220.
21. Suzuki T, Sawaki D, Aizawa K, Munemasa Y, Matsumura T, Ishida J, Nagai R. Kruppel-like factor 5 shows proliferationspecific roles in vascular remodeling, direct stimulation of cell growth, and inhibition of apoptosis. J Biol Chem 2009; 284: 9549-9557.
22. Owens GK, Kumar MS, Wamhoff BR. Molecular regulation of vascular smooth muscle cell differentiation in development and disease. Physiol Rev 2004; 84: 767-801.
23. Wynne BM, Chiao CW, Webb RC. Vascular smooth muscle cell signaling mechanisms for contraction to angiotensin II and endothelin-1. J Am Soc Hypertens 2009; 3: 84-95.
24. Owens GK. Regulation of differentiation of vascular smooth muscle cells. Physiol Rev 1995; 75: 487-517.
25. Lv L, Zhang J, Zhang L, Xue G, Wang P, Meng Q, Liang W. Essential role of Pin1 via STAT3 signalling and mitochondriadependent pathways in restenosis in type 2 diabetes. J Cell Mol Med 2013; 17: 989-1005.
26. Zhang XH, Zheng B, Yang Z, He M, Yue LY, Zhang RN, Zhang M, Zhang W, Zhang X, Wen JK. TMEM16A and myocardin form a positive feedback loop that is disrupted by KLF5 during Ang II-induced vascular remodeling. Hypertension 2015; 66: 412-421.
27. Liao XH, Wang N, Zhao DW, Zheng DL, Zheng L, Xing WJ, Ma WJ, Bao LY, Dong J, Zhang TC. STAT3 protein regulates vascular smooth muscle cell phenotypic switch by interaction with myocardin. J Biol Chem 2015; 290: 19641-19652.