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Apoptosis

, Volume 19, Issue 6, pp 975–983 | Cite as

miR-92a inhibits vascular smooth muscle cell apoptosis: role of the MKK4–JNK pathway

  • Lan Zhang
  • Mi Zhou
  • Yingjie Wang
  • Weibin Huang
  • Gangjian Qin
  • Neal L. Weintraub
  • Yaoliang TangEmail author
Original Paper

Abstract

Vascular smooth muscle cell (VSMC) apoptosis plays an important role in vascular remodeling and atherosclerotic plaque instability. Oxidative stress in diseased vessels promotes VSMC apoptosis in part by activating the c-Jun N-terminal kinase (JNK) pathway, which has been identified as a molecular target of miR-92a in macrophages. Here, we examined the expression and biological activity of miR-92a in VSMC. Quiescent VSMC exhibited a low basal expression of miR-92a, which was positively regulated by serum stimulation and negatively regulated by H2O2. Overexpression of miR-92a decreased H2O2-induced VSMC apoptosis as indicated by TUNEL assay and cleaved caspase-3 protein levels. Using 3′UTR-reporter assay, we found that miR-92a overexpression led to suppression of both mitogen-activated protein kinase kinase 4 (MKK4)- and JNK1-dependent luciferase activity. We also found that 10 mer seed match between miRNA:mRNA pair is more efficient than 8 mer seed match for us to identify authentic miRNA target. Protein levels of active phospho-JNK and phospho-c-Jun, downstream targets of the MKK4–JNK1 pathway, were also decreased by overexpressing miR-92a in VSMC under oxidative stress. Consistent with these findings, overexpression of MKK4 reversed the anti-apoptotic effects of miR-92a in oxidatively stressed VSMC. In conclusion, miR-92a overexpression inhibits H2O2-induced VSMC apoptosis by directly targeting the MKK4–JNK1 pathway.

Keywords

Vascular smooth muscle cells miR-92a JNK Apoptosis Oxidative stress 

Abbreviations

VSMC

Vascular smooth muscle cells

MAPK

Mitogen-activated protein kinase

MKK4

Mitogen-activated protein kinase (MAPK) kinase 4

JNK

C-Jun N-terminal kinase

Notes

Acknowledgments

This work was supported by the American Heart Association Beginning Grant-in-Aid 0765094Y (to Y.T.); NIH Grant HL086555 (to Y.T.), and NIH Grants HL076684 and HL62984 (to N.L.W.).

Conflict of interest

None to disclose.

Supplementary material

10495_2014_987_MOESM1_ESM.pdf (619 kb)
Supplementary Figure 1 VSMC were treated with 100 µM H2O2 in DMEM with 0%, 5%, 10% or 20% FBS for 24 h. TUNEL staining (red) showing representative images (A) and quantitative data (B); nuclei are stained with DAPI (PDF 619 kb)

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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Lan Zhang
    • 1
    • 5
  • Mi Zhou
    • 2
  • Yingjie Wang
    • 3
  • Weibin Huang
    • 1
  • Gangjian Qin
    • 4
  • Neal L. Weintraub
    • 5
  • Yaoliang Tang
    • 5
    Email author
  1. 1.Department of Vascular Surgery, Renji Hospital, School of MedicineShanghai Jiao Tong UniversityShanghaiChina
  2. 2.Department of Cardiac Surgery, Rui Jin Hospital, School of MedicineShanghai Jiao Tong UniversityShanghaiChina
  3. 3.Internal Medicine of Traditional Chinese MedicineShuguang Hospital of Shanghai University of Traditional Chinese MedicineShanghaiChina
  4. 4.Department of Medicine-Cardiology, Feinberg Cardiovascular Research InstituteNorthwestern University Feinberg School of MedicineChicagoUSA
  5. 5.Department of Medicine, Vascular Biology Center, Medical College of GeorgiaGeorgia Regents UniversityAugustaUSA

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