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Participation of miR-200a in TGF-β1-mediated hepatic stellate cell activation

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Abstract

Hepatic stellate cell (HSC) activation is a pivotal event in the initiation and progression of hepatic fibrosis since it mediates transforming growth factor beta 1 (TGF-β1)-driven extracellular matrix (ECM) deposition. MicroRNAs (miRNAs), small non-coding RNAs modulating messenger RNA (mRNA) and protein expression, have emerged as key factors to regulate cell proliferation, differentiation, and apoptosis. Although the function of miR-200a has been discussed in many cancers and fibrotic diseases, its role in hepatic fibrosis is still poorly understood. The aim of this study is to investigate whether miR-200a could attenuate hepatic fibrosis partly through Wnt/β-catenin and TGF-β-dependant mechanisms. Our study found that the expression of endogenous miR-200a was decreased in vitro in TGF-β1-induced HSC activation as well as in vivo in CCl4-induced rat liver fibrosis. Overexpression of miR-200a significantly inhibited α-SMA activity and further affected the proliferation of TGF-β1-dependent activation of HSC. In addition, we identified β-catenin and TGF-β2 as two functional downstream targets for miR-200a. Interestingly, miR-200a specifically suppressed β-catenin in the protein level, whereas miR-200a-mediated suppression of TGF-β2 was shown on both mRNA and protein levels. Our results revealed the critical regulatory role of miR-200a in HSC activation and implied miR-200a as a potential candidate for therapy by deregulation of Wnt/β-catenin and TGFβ signaling pathways, at least in part, via decreasing the expression of β-catenin and TGF-β2.

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Abbreviations

HSC:

Hepatic stellate cell

ECM:

Extracellular matrix

α-SMA:

α-Smooth muscle actin

TGF-β:

Transforming growth factor-β

β-catenin:

Cadherin-associated protein beta

3′-UTR:

3′-Untranslated region

PBS:

Phosphate-buffered saline

SDS:

Sodium dodecyl sulfate

Wt:

Wild type

ZEB:

Zinc-finger E-box-binding homeobox

One-step qRT-PCR:

One-step quantitative real-time PCR

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Acknowledgments

This project was supported by the National Science Foundation of China (Nos: 81072686, 81273526, and 81202978) and the Natural Science Foundation of Anhui Province (KJ2010A178).

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Authors and Affiliations

  1. School of Pharmacy, Anhui Key Laboratory of Bioactivity of Natural Products, Anhui Medical University, Mei Shan Road, Hefei, 230032, Anhui Province, People’s Republic of China

    Xu Sun, Yong He, Tao-Tao Ma, Cheng Huang, Lei Zhang & Jun Li

  2. The Key Laboratory of Anti-inflammatory and Immune Medicine, Ministry of Education, Hefei, People’s Republic of China

    Xu Sun, Yong He, Tao-Tao Ma, Cheng Huang, Lei Zhang & Jun Li

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  1. Xu Sun
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  2. Yong He
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Correspondence to Jun Li.

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Sun, X., He, Y., Ma, TT. et al. Participation of miR-200a in TGF-β1-mediated hepatic stellate cell activation. Mol Cell Biochem 388, 11–23 (2014). https://doi.org/10.1007/s11010-013-1895-0

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