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Research ArticleCellular and Molecular

Role of Vimentin in the Inhibitory Effects of Low-Molecular-Weight Heparin on PC-3M Cell Adhesion to, and Migration through, Endothelium

Yan Pan, Tianluo Lei, Bao Teng, Jihong Liu, Jianzhao Zhang, Yu An, Yuan Xiao, Jing Han, Xueyang Pan, Junhua Wang, Heming Yu, Hong Ren and Xuejun Li
Journal of Pharmacology and Experimental Therapeutics October 2011, 339 (1) 82-92; DOI: https://doi.org/10.1124/jpet.111.182055
Yan Pan
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Tianluo Lei
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Bao Teng
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Jihong Liu
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Jianzhao Zhang
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Yu An
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Yuan Xiao
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Jing Han
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Xueyang Pan
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Junhua Wang
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Heming Yu
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Hong Ren
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Xuejun Li
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Abstract

Low-molecular-weight heparin (LMWH) has been used in cancer patients with venous thromboembolic complications, resulting in a higher survival rate and an inhibitory action on experimental metastasis. In the present study, human umbilical vein endothelial cells (HUVECs) were treated with LMWH for 24 h. We found that the resulting HUVECs could significantly inhibit the highly metastatic human prostate cancer cell line (PC-3M) in terms of its adhesion to the endothelium and migration across the endothelium, according to scanning electron microscopy. We also determined the elevated levels of endothelial intercellular Ca2+ concentration after the adhesion of PC-3M cells to HUVECs was greatly reduced by incubation with LMWH. Using proteomics, we surveyed the global protein changes in HUVECs after LMWH treatment and identified four down-regulated proteins that were possible isoforms of cytoskeletal vimentin intermediate filaments, cartilage-derived C-type lectin, and serine/threonine protein phosphatase 1β (PP-1B). LMWH affected the morphology of vimentin and the expression levels of αv integrin and PP-1B in HUVECs bound to PC-3M cells. Vimentin assists in the adhesion of PC-3M cells, which was confirmed by short interfering RNA experiments. Furthermore, the direct binding of purified vimentin protein with LMWH was detected with surface plasmon resonance methods. However, when we used fluorescence-labeled heparin for 24 h to identify whether this binding occurred within cells, heparin was distributed principally around endothelial cells. Taken together, these findings suggest that the monoincubation of LMWH with HUVECs could inhibit PC-3M cell adhesion to, and migration through, endothelium. LMWH's regulation of vimentin plays a role in the antimetastatic action.

Footnotes

  • This work was supported by the National Natural Science Foundation of China [Grants 81020108031, 30572202, 30973558, 30772571, 30901803, 30901815]; the Major Specialized Research Fund from the Ministry of Science and Technology in China [Grant 2009ZX09103-144]; the Research Fund from Ministry of Education of China (111 Projects) [Grant B07001]; and the Scientific Research Foundation for the Returned Overseas Chinese Scholars, State Education Ministry of China.

  • Article, publication date, and citation information can be found at http://jpet.aspetjournals.org.

    doi:10.1124/jpet.111.182055.

  • ABBREVIATIONS:

    LMWH
    low-molecular-weight heparin
    CFDA
    carboxyfluorescein diacetate
    DTT
    dithiothreitol
    EC
    endothelial cell
    HUVEC
    human umbilical vein endothelial cell
    IEF
    isoelectric focusing
    IPG
    immobilized pH gradient
    MALDI-TOF-MS
    matrix-assisted laser desorption ionization time-of-flight mass spectrometry
    PBS
    phosphate-buffered saline
    RU
    response units
    PAGE
    polyacrylamide gel electrophoresis
    SEM
    scanning electron microscopy
    siRNA
    short interfering RNA
    SPR
    surface plasmon resonance
    TGF-β
    transforming growth factor-β
    PP-1B
    protein phosphatase 1β
    2D
    two-dimensional
    FITC
    fluorescein isothiocyanate
    RT-PCR
    reverse transcription-polymerase chain reaction; [Ca2+]i intracellular calcium concentration
    ns-siRNA
    nonsilencing siRNA
    siVimentin
    vimentin-targeted siRNA.

  • Received March 28, 2011.
  • Accepted June 30, 2011.
  • Copyright © 2011 by The American Society for Pharmacology and Experimental Therapeutics
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Journal of Pharmacology and Experimental Therapeutics: 339 (1)
Journal of Pharmacology and Experimental Therapeutics
Vol. 339, Issue 1
1 Oct 2011
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Research ArticleCellular and Molecular

Regulation of Vimentin by LMWH Inhibits Cell Migration

Yan Pan, Tianluo Lei, Bao Teng, Jihong Liu, Jianzhao Zhang, Yu An, Yuan Xiao, Jing Han, Xueyang Pan, Junhua Wang, Heming Yu, Hong Ren and Xuejun Li
Journal of Pharmacology and Experimental Therapeutics October 1, 2011, 339 (1) 82-92; DOI: https://doi.org/10.1124/jpet.111.182055

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Research ArticleCellular and Molecular

Regulation of Vimentin by LMWH Inhibits Cell Migration

Yan Pan, Tianluo Lei, Bao Teng, Jihong Liu, Jianzhao Zhang, Yu An, Yuan Xiao, Jing Han, Xueyang Pan, Junhua Wang, Heming Yu, Hong Ren and Xuejun Li
Journal of Pharmacology and Experimental Therapeutics October 1, 2011, 339 (1) 82-92; DOI: https://doi.org/10.1124/jpet.111.182055
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