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Des-γ-carboxy prothrombin stimulates human vascular endothelial cell growth and migration

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Abstract

Des-γ-carboxy prothrombin (DCP) is an aberrant prothrombin produced by hepatocellular carcinoma (HCC) cells. Serum and tissue DCP expressions are thought to reflect the biological malignant potential of HCC. However, the role of DCP in the development of angiogenesis is not well understood. Herein, we report the effects of DCP on growth and migration of human vascular endothelial cells. DCP significantly stimulated the proliferation of HUVEC (ECV304) cells in a dose and time dependent manner, as measured by the MTT assay. A continuous rapid migration of ECV304 cells was observed in the presence of DCP measured by the scratch wound assay. The continuous rapid invasive activity, measured by transwell chamber assay also showed that DCP increased endothelial cells migration through the reconstituted extracellular matrix (Matrigel). Further, the tube formation of vascular endothelial cells on 3-D Matrigel showed an increased number of branch points of ECV304 cells induced by DCP in a dose dependent manner. The levels of vascular endothelial cell growth-related angiogenic factors and matrix metalloproteinase were also examined. DCP significantly stimulated the expression levels of epidermal growth factor receptor (EGFR), vascular endothelial growth factor (VEGF), and matrix metalloproteinase (MMP)-2 (latent and active). Together, these data suggest that DCP is a novel type of vascular endothelial growth factor that possesses potent mitogenic and migrative activities in angiogenesis of HCC.

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Abbreviations

HCC:

Hepatocellular carcinoma

DCP:

Des-γ-carboxy prothrombin

HUVEC:

Human umbilical vein vascular endothelial cell line

MTT:

3-[4,5-Dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide

PBS:

Phosphate buffered saline

EDTA:

Ethylenediamine tetraacetic acid

HEPES:

Hydroethyl piperazine-ethane sulfonic acid

ELISA:

Enzyme-link immunosorbent assay

EGFR:

Epidermal growth factor receptor

VEGF:

Vascular endothelial growth factor

MMPs:

Matrix metalloproteinase

ECM:

Extracellular matrix

HGF:

Hepatocyte growth factor

KDR:

Kinase insert domain receptor

MAPK:

Mitogen-activated protein kinase

TGF-β:

Transforming growth factor-β

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Acknowledgments

This project was supported by the Natural Science Foundation of China, Shandong Provincial Foundation for Natural Science (Y2006C87) and Shandong University Student’s Innovation Program. This work was also supported in part by Grant-in-Aid from the Ministry of Education, Science, Sports and Culture of Japan (20015009). We thank Dr M. D. Lloyd (University of Bath, U. K.) for critically reading this manuscript.

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Authors and Affiliations

  1. School of Ocean Sciences, Shandong University at Weihai, 264209, Weihai, China

    Su-Bo Wang

  2. Department of Pharmacology, School of Pharmaceutical Sciences, Shandong University, 250012, Jinan, China

    Yan-Na Cheng, Li-Rui Sun, Ming-Hui Chen & Xian-Jun Qu

  3. Department of Pharmacology, Institute of Materia Medica, Shandong Academy of Medical Sciences, 250062, Jinan, China

    Shu-Xiang Cui

  4. Department of Pharmacy and Pharmacology, University of Bath, Claverton Down, Bath, BA2, 7AV, UK

    Julia Li Zhong & S. G. Ward

  5. Hepato-Biliary-Pancreatic Surgery Division, Department of Surgery, Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, 113-8655, Japan

    Norihiro Kokudo & Wei Tang

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  1. Su-Bo Wang
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Correspondence to Xian-Jun Qu.

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S. Cui and X.-J. Qu contributed equally to this work.

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Wang, SB., Cheng, YN., Cui, SX. et al. Des-γ-carboxy prothrombin stimulates human vascular endothelial cell growth and migration. Clin Exp Metastasis 26, 469–477 (2009). https://doi.org/10.1007/s10585-009-9246-y

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  • DOI: https://doi.org/10.1007/s10585-009-9246-y

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