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

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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.

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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

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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.

Author information

Authors and Affiliations

  1. Department of Vascular Surgery, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, 200127, China

    Lan Zhang & Weibin Huang

  2. Department of Cardiac Surgery, Rui Jin Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, 200025, China

    Mi Zhou

  3. Internal Medicine of Traditional Chinese Medicine, Shuguang Hospital of Shanghai University of Traditional Chinese Medicine, Shanghai, 201203, China

    Yingjie Wang

  4. Department of Medicine-Cardiology, Feinberg Cardiovascular Research Institute, Northwestern University Feinberg School of Medicine, Chicago, IL, USA

    Gangjian Qin

  5. Department of Medicine, Vascular Biology Center, Medical College of Georgia, Georgia Regents University, 1459 Laney Walker Blvd, Augusta, GA, 30912, USA

    Lan Zhang, Neal L. Weintraub & Yaoliang Tang

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  1. Lan Zhang
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Correspondence to Yaoliang Tang.

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10495_2014_987_MOESM1_ESM.pdf

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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Zhang, L., Zhou, M., Wang, Y. et al. miR-92a inhibits vascular smooth muscle cell apoptosis: role of the MKK4–JNK pathway. Apoptosis 19, 975–983 (2014). https://doi.org/10.1007/s10495-014-0987-y

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  • DOI: https://doi.org/10.1007/s10495-014-0987-y

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