microRNA-214-3p在周期性张应变诱导内皮祖细胞分化和增殖中的作用
|
摘要
目的探讨microRNA-214-3p(miR-214-3p)在周期性张应变诱导内皮祖细胞(endothelial progenitor cells, EPCs)分化和增殖中的作用。方法采用FX-5000T细胞周期性张应变加载装置对EPCs施加生理水平的周期性张应变(5%幅度、1.25 Hz频率),加载时间24 h。应用miRNAs芯片筛选周期性张应变调控下差异表达的miRNAs,并挑选miR-214-3p进行深入研究。实时荧光定量PCR方法检测EPCs内平滑肌细胞(vascular smooth muscle cells, VSMCs)相标志分子的表达,BrdU结合酶联免疫吸附ELISA法检测EPCs增殖功能。之后,使用miR-214-3p抑制剂抑制miR-214-3p的表达,检测EPC内VSMC相标志分子表达及EPCs增殖。结果周期性张应变显著抑制miR-214-3p表达,并抑制EPCs向VSMC相分化,同时显著促进EPCs增殖。在静态条件下,使用miR-214-3p抑制剂干扰miR-214-3p的表达,miR-214-3p水平下降同样会抑制EPCs向VSMC相分化,并且诱导EPCs增殖能力显著上升。结论生理水平的周期性张应变能够抑制EPCs内miR-214-3p表达,从而抑制EPCs向VSMC相分化,并且促进EPCs增殖。研究结果为血管损伤的治疗提供新的治疗靶点。
Objective To investigate the role of microRNA-214-3p(miR-214-3p) in differentiation and proliferation of endothelial progenitor cells(EPCs) induced by cyclic stretch. Methods EPCs were exposed to cyclic stretch at physiological level(with the magnitude of 5%, at a constant frequency of 1.25 Hz) for 24 h by FX-5000T Strain Unit. miRNAs array was performed to identify the expression profiling of miRNAs. Real-time PCR was used to examine the expression levels of miRs. The expression of vascular smooth muscle cells(VSMCs) markers in EPCs was detected by real-time PCR. EPC proliferation was detected by BrdU ELISA assay. After EPCs were transfected with miR-214-3p inhibitor(IN) to knockdown expression of miR-214-3p, the level of VSMC markers expression and EPC proliferation was detected. Results Cyclic stretch significantly decreased miR-214-3p expression, depressed EPC differentiation toward VSMCs, and increased EPCs proliferation. Similarly, transfection with the miR-214-3p inhibitor led to the decreased expression of VSMC markers under static station. Meanwhile, miR-214-3p down-regulation promoted EPC proliferation significantly. Conclusions Physiological cyclic stretch could down-regulate the expression of miR-214-3p in EPCs, depress EPC differentiation towards VSMC and promote EPC proliferation eventually. Therefore, the research findings provide a potential therapeutic strategy for treating vessel injuries.
Objective To investigate the role of microRNA-214-3p(miR-214-3p) in differentiation and proliferation of endothelial progenitor cells(EPCs) induced by cyclic stretch. Methods EPCs were exposed to cyclic stretch at physiological level(with the magnitude of 5%, at a constant frequency of 1.25 Hz) for 24 h by FX-5000T Strain Unit. miRNAs array was performed to identify the expression profiling of miRNAs. Real-time PCR was used to examine the expression levels of miRs. The expression of vascular smooth muscle cells(VSMCs) markers in EPCs was detected by real-time PCR. EPC proliferation was detected by BrdU ELISA assay. After EPCs were transfected with miR-214-3p inhibitor(IN) to knockdown expression of miR-214-3p, the level of VSMC markers expression and EPC proliferation was detected. Results Cyclic stretch significantly decreased miR-214-3p expression, depressed EPC differentiation toward VSMCs, and increased EPCs proliferation. Similarly, transfection with the miR-214-3p inhibitor led to the decreased expression of VSMC markers under static station. Meanwhile, miR-214-3p down-regulation promoted EPC proliferation significantly. Conclusions Physiological cyclic stretch could down-regulate the expression of miR-214-3p in EPCs, depress EPC differentiation towards VSMC and promote EPC proliferation eventually. Therefore, the research findings provide a potential therapeutic strategy for treating vessel injuries.
引文
[1] KULISZEWSKI MA, KOBULNIK J, LINDNER JR, et al. Vascular gene transfer of SDF-1 promotes endothelial progenitor cell engraftment and enhances angiogenesis in ischemic muscle [J]. Mol Ther, 2011, 19(5): 895-902.
[2] DOYLE B, METHAROM P, CAPLICE NM. Endothelial progenitor cells [J]. Endothelium, 2006, 13(6): 403-410.
[3] ANTóNIO N, FERNANDES R, SOARES A, et al. Reduced levels of circulating endothelial progenitor cells in acute myocardial infarction patients with diabetes or pre-diabetes: Accompanying the glycemic continuum [J]. Cardiovasc Diabetol, 2014, 13(1): 101.
[4] WERNER N, KOSIOL S, SCHIEGL T, et al. Circulating endothelial progenitor cells and cardiovascular outcomes [J].New Engl J Med, 2005, 353(10): 999-1007.
[5] VASA M, FICHTLSCHERER S, AICHER A, et al. Number and migratory activity of circulating endothelial progenitor cells inversely correlate with risk factors for coronary artery disease [J]. Circ Res, 2001, 89(1): e1-e7.
[6] HILL JM, ZALOS G, HALCOX JPJ, et al. Circulating endothelial progenitor cells, vascular function, and cardiovascular risk [J]. New Engl J Med, 2003, 348(7): 593-600.
[7] CHEN MC, CHEN CJ, YANG CH, et al. Relationship of the percentage of circulating endothelial progenitor cell to the severity of coronary artery disease [J]. Heart Vessels, 2008, 23(1): 47-52.
[8] ZARGHAM R. Preventing restenosis after angioplasty: A multistage approach [J]. Clin Sci, 2008, 114(4): 257-264.
[9] ZHANG C, ZENG L, EMANUELI C, et al. Blood flow and stem cells in vascular disease [J]. Cardiovasc Res, 2013, 99(2): 251-259.
[10] INUI M, MARTELLO G, PICCOLO S. MicroRNA control of signal transduction [J]. Nat Rev Mol Cell Bio, 2010, 11(4): 252-263.
[11] FABIAN MR, SONENBERG N, FILIPOWICZ W. Regulation of mRNA translation and stability by microRNAs [J]. Annu Rev Biochem, 2010, 79: 351-379.
[12] GAO Y, FENG B, HAN S, et al. MicroRNA-129 in human cancers: From tumorigenesis to clinical treatment [J]. Cell Physiol Biochem, 2016, 39(6): 2186-2202.
[13] 马英英, 王璐, 包晗, 等. microRNA-133b 在低切应力诱导血管内皮细胞影响血管平滑肌细胞增殖中的作用[J]. 医用生物力学, 2016, 31 (5): 408-415.MA YY, WANG L, BAO H, et al. The role of microRNA-133b in proliferation of vascular smooth muscle cells induced by endothelial cells under low shear stress [J]. J Med Biomech, 2016, 31 (5): 408-415.
[14] KANE NM, THRASHER AJ, ANGELINI GD, et al. Concise review: MicroRNAs as modulators of stem cells and angiogenesis [J]. Stem Cells, 2014, 32(5): 1059-1066.
[15] LIN CY, TZENG HE, LI TM, et al. WISP-3 inhibition of miR-452 promotes VEGF-A expression in chondrosarcoma cells and induces endothelial progenitor cells angiogenesis [J]. Oncotarget, 2017, 8(24): 39571-39581
[16] MENG S, CAO JT, WANG LS, et al. MicroRNA 107 partly inhibits endothelial progenitor cells differentiation via HIF-1β [J]. PloS One, 2012, 7(7): e40323.
[17] LEI Z, MIL A, BRANDT MM, et al. MicroRNA-132/212 family enhances arteriogenesis after hindlimb ischaemia through modulation of the Ras-MAPK pathway [J]. J Cell Mol Med, 2015, 19(8): 1994-2005.
[18] WANG L, BAO H, WANG KX, et al. Secreted miR-27a induced by cyclic stretch modulates the proliferation of endothelial cells in hypertension via GRK6 [J]. Sci Rep, 2017, doi: 10.1038/srep41058.
[19] MONDADORI DOS SANTOS A, METZINGER L, HADDAD O, et al. miR-126 is involved in vascular remodeling under laminar shear stress [J]. Biomed Res Int, 2015, doi: 10.1155/2015/497280.
[20] CHENG BB, QU MJ, WU LL, et al. MicroRNA-34a targets Forkhead box j2 to modulate differentiation of endothelial progenitor cells in response to shear stress [J]. J Mol Cell Cardiol, 2014, doi: 10.1016/j.yjmcc.2014.04.016.
[21] CUI X, ZHANG X, GUAN X, et al. Shear stress augments the endothelial cell differentiation marker expression in late EPCs by upregulating integrins [J]. Biochem Bioph Res Co, 2012, 425(2): 419-425.
[22] ASAHARA T, MUROHARA T, SULLIVAN A, et al. Isolation of putative progenitor endothelial cells for angiogenesis [J]. Science, 1997, 275(5302): 964-966.
[23] LI R, LIANG L, DOU Y, et al. Mechanical strain regulates osteogenic and adipogenic differentiation of bone marrow mesenchymal stem cells [J]. Biomed Res Int, 2015, doi: 10.1155/2015/873251.
[24] LOHBERGER B, KALTENEGGER H, STUENDL N, et al. Effect of cyclic mechanical stimulation on the expression of osteogenesis genes in human intraoral mesenchymal stromal and progenitor cells [J]. Biomed Res Int, 2014, doi: 10.1155/2014/189516.
[25] BERGER S, LAVIE L. Endothelial progenitor cells in cardiovascular disease and hypoxia-potential implications to obstructive sleep apnea [J]. Transl Res, 2011, 158(1): 1-13.
[26] GEORGE AL, BANGALORE-PRAKASH P, RAJORIA S, et al. Endothelial progenitor cell biology in disease and tissue regeneration [J]. J Hematol Oncol, 2011, doi: 10.1186/1756-8722-4-24.
[27] BADR G, HOZZEIN WN, BADR BM, et al. Bee venom accelerates wound healing in diabetic mice by suppressing activating transcription factor-3 (ATF-3) and inducible nitric oxide synthase (iNOS)-mediated oxidative stress and recruiting bone marrow-derived endothelial progenitor cells [J]. J Cell Physiol, 2016, 231(10): 2159-2171.
[28] LERMAN A. Restenosis [J]. Circulation, 2005, 111: 8-10.
[29] CHENG BB, YAN ZQ, YAO QP, et al. Association of SIRT1 expression with shear stress induced endothelial progenitor cell differentiation [J].J Cell Biochem, 2012, 113(12): 3663-3671.
[30] BRIASOULIS A, TOUSOULIS D, ANTONIADES C, et al. The role of endothelial progenitor cells in vascular repair after arterial injury and atherosclerotic plaque development [J]. Cardiovasc Ther, 2011, 29(2): 125-139.
[31] XIA WH, YANG Z, XU SY, et al. Age-related decline in reendothelialization capacity of human endothelial progenitor cells is restored by shear stress [J]. Hypertension, 2012, 59(6): 1225-1231.
[32] ABIKO H, FUJIWARA S, OHASHI K, et al. Rho guanine nucleotide exchange factors involved in cyclic-stretch-induced reorientation of vascular endothelial cells [J]. J Cell Sci, 2015, 128(9): 1683-1695.
[33] MANTELLA LE, SINGH KK, SANDHU P, et al. Fingerprint of long non-coding RNA regulated by cyclic mechanical stretch in human aortic smooth muscle cells: Implications for hypertension [J]. Mol Cell Biochem, 2017, 435(1-2): 163-173.
[34] ZUO K, ZHI K, ZHANG X, et al. A dysregulated microRNA-26a/EphA2 axis impairs endothelial progenitor cell function via the p38 MAPK/VEGF pathway [J]. Cell Physiol Biochem, 2015, 35(2): 477-488.
[35] YE M, LI D, YANG J, et al. MicroRNA-130a targets MAP3K12 to modulate diabetic endothelial progenitor cell function [J]. Cell Physiol Biochem, 2015, 36(2): 712-726.
[36] WEI Y, NAZARI-JAHANTIGH M, NETH P, et al. MicroRNA-126,-145, and-155 [J]. Arterioscl Throm Vas, 2013, 33(3): 449-454.
[37] SPINETTI G, FORTUNATO O, CAPORALI A, et al. MicroRNA-15a and MicroRNA-16 impair human circulating pro-angiogenic cell (PAC) functions and are increased in the PACs and serum of patients with critical limb ischemia [J]. Circ Res, 2013, 112(2): 335-346.
[38] SYED DN, LALL RK, MUKHTAR H. MicroRNAs and photocarcinogenesis [J]. Photochem Photobiol, 2015, 91(1): 173-187.
[39] LIU J, LI D, DANG L, et al. Osteoclastic miR-214 targets TRAF3 to contribute to osteolytic bone metastasis of breast cancer [J]. Sci Rep, 2017, doi: 10.1038/srep40487.
[40] LI Y, LI Y, CHEN Y, et al. MicroRNA-214-3p inhibits proliferation and cell cycle progression by targeting MELK in hepatocellular carcinoma and correlates cancer prognosis [J]. Cancer Cell Int, 2017, doi: 10.1186/s12935-017-0471-1.
[2] DOYLE B, METHAROM P, CAPLICE NM. Endothelial progenitor cells [J]. Endothelium, 2006, 13(6): 403-410.
[3] ANTóNIO N, FERNANDES R, SOARES A, et al. Reduced levels of circulating endothelial progenitor cells in acute myocardial infarction patients with diabetes or pre-diabetes: Accompanying the glycemic continuum [J]. Cardiovasc Diabetol, 2014, 13(1): 101.
[4] WERNER N, KOSIOL S, SCHIEGL T, et al. Circulating endothelial progenitor cells and cardiovascular outcomes [J].New Engl J Med, 2005, 353(10): 999-1007.
[5] VASA M, FICHTLSCHERER S, AICHER A, et al. Number and migratory activity of circulating endothelial progenitor cells inversely correlate with risk factors for coronary artery disease [J]. Circ Res, 2001, 89(1): e1-e7.
[6] HILL JM, ZALOS G, HALCOX JPJ, et al. Circulating endothelial progenitor cells, vascular function, and cardiovascular risk [J]. New Engl J Med, 2003, 348(7): 593-600.
[7] CHEN MC, CHEN CJ, YANG CH, et al. Relationship of the percentage of circulating endothelial progenitor cell to the severity of coronary artery disease [J]. Heart Vessels, 2008, 23(1): 47-52.
[8] ZARGHAM R. Preventing restenosis after angioplasty: A multistage approach [J]. Clin Sci, 2008, 114(4): 257-264.
[9] ZHANG C, ZENG L, EMANUELI C, et al. Blood flow and stem cells in vascular disease [J]. Cardiovasc Res, 2013, 99(2): 251-259.
[10] INUI M, MARTELLO G, PICCOLO S. MicroRNA control of signal transduction [J]. Nat Rev Mol Cell Bio, 2010, 11(4): 252-263.
[11] FABIAN MR, SONENBERG N, FILIPOWICZ W. Regulation of mRNA translation and stability by microRNAs [J]. Annu Rev Biochem, 2010, 79: 351-379.
[12] GAO Y, FENG B, HAN S, et al. MicroRNA-129 in human cancers: From tumorigenesis to clinical treatment [J]. Cell Physiol Biochem, 2016, 39(6): 2186-2202.
[13] 马英英, 王璐, 包晗, 等. microRNA-133b 在低切应力诱导血管内皮细胞影响血管平滑肌细胞增殖中的作用[J]. 医用生物力学, 2016, 31 (5): 408-415.MA YY, WANG L, BAO H, et al. The role of microRNA-133b in proliferation of vascular smooth muscle cells induced by endothelial cells under low shear stress [J]. J Med Biomech, 2016, 31 (5): 408-415.
[14] KANE NM, THRASHER AJ, ANGELINI GD, et al. Concise review: MicroRNAs as modulators of stem cells and angiogenesis [J]. Stem Cells, 2014, 32(5): 1059-1066.
[15] LIN CY, TZENG HE, LI TM, et al. WISP-3 inhibition of miR-452 promotes VEGF-A expression in chondrosarcoma cells and induces endothelial progenitor cells angiogenesis [J]. Oncotarget, 2017, 8(24): 39571-39581
[16] MENG S, CAO JT, WANG LS, et al. MicroRNA 107 partly inhibits endothelial progenitor cells differentiation via HIF-1β [J]. PloS One, 2012, 7(7): e40323.
[17] LEI Z, MIL A, BRANDT MM, et al. MicroRNA-132/212 family enhances arteriogenesis after hindlimb ischaemia through modulation of the Ras-MAPK pathway [J]. J Cell Mol Med, 2015, 19(8): 1994-2005.
[18] WANG L, BAO H, WANG KX, et al. Secreted miR-27a induced by cyclic stretch modulates the proliferation of endothelial cells in hypertension via GRK6 [J]. Sci Rep, 2017, doi: 10.1038/srep41058.
[19] MONDADORI DOS SANTOS A, METZINGER L, HADDAD O, et al. miR-126 is involved in vascular remodeling under laminar shear stress [J]. Biomed Res Int, 2015, doi: 10.1155/2015/497280.
[20] CHENG BB, QU MJ, WU LL, et al. MicroRNA-34a targets Forkhead box j2 to modulate differentiation of endothelial progenitor cells in response to shear stress [J]. J Mol Cell Cardiol, 2014, doi: 10.1016/j.yjmcc.2014.04.016.
[21] CUI X, ZHANG X, GUAN X, et al. Shear stress augments the endothelial cell differentiation marker expression in late EPCs by upregulating integrins [J]. Biochem Bioph Res Co, 2012, 425(2): 419-425.
[22] ASAHARA T, MUROHARA T, SULLIVAN A, et al. Isolation of putative progenitor endothelial cells for angiogenesis [J]. Science, 1997, 275(5302): 964-966.
[23] LI R, LIANG L, DOU Y, et al. Mechanical strain regulates osteogenic and adipogenic differentiation of bone marrow mesenchymal stem cells [J]. Biomed Res Int, 2015, doi: 10.1155/2015/873251.
[24] LOHBERGER B, KALTENEGGER H, STUENDL N, et al. Effect of cyclic mechanical stimulation on the expression of osteogenesis genes in human intraoral mesenchymal stromal and progenitor cells [J]. Biomed Res Int, 2014, doi: 10.1155/2014/189516.
[25] BERGER S, LAVIE L. Endothelial progenitor cells in cardiovascular disease and hypoxia-potential implications to obstructive sleep apnea [J]. Transl Res, 2011, 158(1): 1-13.
[26] GEORGE AL, BANGALORE-PRAKASH P, RAJORIA S, et al. Endothelial progenitor cell biology in disease and tissue regeneration [J]. J Hematol Oncol, 2011, doi: 10.1186/1756-8722-4-24.
[27] BADR G, HOZZEIN WN, BADR BM, et al. Bee venom accelerates wound healing in diabetic mice by suppressing activating transcription factor-3 (ATF-3) and inducible nitric oxide synthase (iNOS)-mediated oxidative stress and recruiting bone marrow-derived endothelial progenitor cells [J]. J Cell Physiol, 2016, 231(10): 2159-2171.
[28] LERMAN A. Restenosis [J]. Circulation, 2005, 111: 8-10.
[29] CHENG BB, YAN ZQ, YAO QP, et al. Association of SIRT1 expression with shear stress induced endothelial progenitor cell differentiation [J].J Cell Biochem, 2012, 113(12): 3663-3671.
[30] BRIASOULIS A, TOUSOULIS D, ANTONIADES C, et al. The role of endothelial progenitor cells in vascular repair after arterial injury and atherosclerotic plaque development [J]. Cardiovasc Ther, 2011, 29(2): 125-139.
[31] XIA WH, YANG Z, XU SY, et al. Age-related decline in reendothelialization capacity of human endothelial progenitor cells is restored by shear stress [J]. Hypertension, 2012, 59(6): 1225-1231.
[32] ABIKO H, FUJIWARA S, OHASHI K, et al. Rho guanine nucleotide exchange factors involved in cyclic-stretch-induced reorientation of vascular endothelial cells [J]. J Cell Sci, 2015, 128(9): 1683-1695.
[33] MANTELLA LE, SINGH KK, SANDHU P, et al. Fingerprint of long non-coding RNA regulated by cyclic mechanical stretch in human aortic smooth muscle cells: Implications for hypertension [J]. Mol Cell Biochem, 2017, 435(1-2): 163-173.
[34] ZUO K, ZHI K, ZHANG X, et al. A dysregulated microRNA-26a/EphA2 axis impairs endothelial progenitor cell function via the p38 MAPK/VEGF pathway [J]. Cell Physiol Biochem, 2015, 35(2): 477-488.
[35] YE M, LI D, YANG J, et al. MicroRNA-130a targets MAP3K12 to modulate diabetic endothelial progenitor cell function [J]. Cell Physiol Biochem, 2015, 36(2): 712-726.
[36] WEI Y, NAZARI-JAHANTIGH M, NETH P, et al. MicroRNA-126,-145, and-155 [J]. Arterioscl Throm Vas, 2013, 33(3): 449-454.
[37] SPINETTI G, FORTUNATO O, CAPORALI A, et al. MicroRNA-15a and MicroRNA-16 impair human circulating pro-angiogenic cell (PAC) functions and are increased in the PACs and serum of patients with critical limb ischemia [J]. Circ Res, 2013, 112(2): 335-346.
[38] SYED DN, LALL RK, MUKHTAR H. MicroRNAs and photocarcinogenesis [J]. Photochem Photobiol, 2015, 91(1): 173-187.
[39] LIU J, LI D, DANG L, et al. Osteoclastic miR-214 targets TRAF3 to contribute to osteolytic bone metastasis of breast cancer [J]. Sci Rep, 2017, doi: 10.1038/srep40487.
[40] LI Y, LI Y, CHEN Y, et al. MicroRNA-214-3p inhibits proliferation and cell cycle progression by targeting MELK in hepatocellular carcinoma and correlates cancer prognosis [J]. Cancer Cell Int, 2017, doi: 10.1186/s12935-017-0471-1.