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Myocyte enhancer factor 2C regulation of hepatocellular carcinoma via vascular endothelial growth factor and Wnt/β-catenin signaling

Oncogene volume 34, pages 4089–4097 (2015)Cite this article

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Abstract

Hepatocellular carcinoma (HCC) is one of the leading malignancies worldwide. Myocyte enhancer factor 2C (MEF2C) was traditionally regarded as a development-associated factor and was recently reported to be an oncogene candidate. We have previously reported overexpression of MEF2C in HCC; however, the roles of MEF2C in HCC remain to be clarified. In this study, HCC cell lines and a xenograft mouse model were used to determine the functions of MEF2C in vitro and in vivo, respectively. Specific plasmids and small interfering RNA were used to upregulate and downregulate MEF2C expression, respectively. Functional assays were performed to assess the influence of MEF2C on cell proliferation, and VEGF-induced vasculogenic mimicry, migration/invasion as well as angiogenesis. Co-immunoprecipitation was conducted to identify the interaction of MEF2C and β-catenin. Human HCC tissue microarrays were used to investigate correlations among MEF2C, β-catenin and involved biomarkers. MEF2C was found to mediate VEGF-induced vasculogenic mimicry, angiogenesis and migration/invasion, with involvement of the p38 MAPK and PKC signaling pathways. However, MEF2C itself inhibited tumor growth in vitro and in vivo. MEF2C was upregulated by and directly interacted with β-catenin. The nuclear translocation of β-catenin blocked by MEF2C was responsible for MEF2C-mediated growth inhibition. The nuclear translocation of MEF2C was associated with intracellular calcium signaling induced by β-catenin. HCC microarrays showed correlations of nuclear MEF2C with the angiogenesis-associated biomarker, CD31, and cytosolic MEF2C with the proliferation-associated biomarker, Ki-67. MEF2C showed double-edged activities in HCC, namely mediating VEGF-induced malignancy enhancement while inhibiting cancer proliferation via blockade of Wnt/β-catenin signaling. The overall effect of MEF2C in HCC progression regulation was dictated by its subcellular distribution. This should be determined prior to any MEF2C-associated intervention in HCC.

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Acknowledgements

We thank Professor Xu Qiang (State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, China) for the generous gift of MEF2C overexpression plasmid. We appreciate Mr Xie Shangzhi, Mr Hu Liqiang, and Miss Chen Conglin (The Second Affiliated Hospital, Zhejiang University School of Medicine, China) for their great help in certain experiments. This study was financially supported by the National Natural Science Foundation of China (No. 81171884), the National Key Basic Research Program of China (No. 2014CB542101), the Ministry-Province Co-supportive Project of China, and Innovation and High-Level Talent Training Program of Department of Health of Zhejiang.

Author Contributions

XLB, QZ, ZZP and TBL designed the experiments. XLB, QZ, LYY, FL, XS, YC, QDH, QHF, ZC and WS performed the experiments and analyzed the data. TBL supervised the project. XLB, QZ and TBL wrote the report.

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Author notes

  1. X L Bai and Q Zhang: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Hepatobiliary and Pancreatic Surgery, The Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China

    X L Bai, Q Zhang, L Y Ye, X Sun, Q D Hu, Q H Fu, W Su & T B Liang

  2. Key Laboratory of Cancer Prevention and Intervention, The Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China

    X L Bai, Q Zhang, L Y Ye & T B Liang

  3. Department of Neurosurgery, The Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China

    F Liang

  4. Department of General Surgery, The Children’s Hospital, Zhejiang University School of Medicine, Hangzhou, China

    Y Chen

  5. Zhejiang Key Laboratory of Gastro-Intestinal Pathophysiology, Zhejiang Hospital of Traditional Chinese Medicine, Zhejiang Chinese Medical University, Hangzhou, China

    Z Chen

  6. National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, MD, USA

    Z P Zhuang

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  1. X L Bai
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Correspondence to T B Liang.

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Bai, X., Zhang, Q., Ye, L. et al. Myocyte enhancer factor 2C regulation of hepatocellular carcinoma via vascular endothelial growth factor and Wnt/β-catenin signaling. Oncogene 34, 4089–4097 (2015). https://doi.org/10.1038/onc.2014.337

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