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Human tissue kallikrein 1 gene delivery inhibits PDGF-BB-induced vascular smooth muscle cells proliferation and upregulates the expressions of p27Kip1 and p2lCip1

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Abstract

Tissue kallikrein 1 cleaves kininogen substrate to produce vasoactive kinin peptides that have been implicated in inhibiting neointimal hyperplasia in rat carotid arteries after balloon injury. However, its effects on the proliferation, cell cycle and its mechanisms, for example, cyclin-dependent kinase inhibitors, p27Kip1 and p2lCip1 in vascular biology are poorly understood. The objective of this study was to explore the effects of human tissue kallikrein 1 (hTK1) mediated by recombinant adenovirus (Ad-hTK1) on proliferation and cell cycle of vascular smooth muscle cells (VSMCs) derived from spontaneously hypertensive rats induced by platelet-derived growth factor-BB (PDGF-BB) in vitro. The results showed that, within a given multiplicity of infection (MOI) and time, the hTK1 gene delivery inhibited PDGF-BB-stimulating VSMCs growth in a concentration-dependent (20–100 MOI) and time-dependent (2–5 days) manner by cell counting, with a peak inhibition rate at 36.3% at 72 h (P < 0.01). In addition, hTK1 gene delivery significantly suppressed PDGF-BB-induced proliferation of VSMCs by methyl thiazolyl tetrazoliuin assay, and decreased the percentage of cells in the S phase and in DNA synthesis by flow cytometry, with a peak inhibition rate at 30.2 and 36.4%, respectively (P < 0.01). Western blot assay showed that the protein levels of p27Kip1 and p2lCip1 in cells infected with Ad-hTK1 were much more abundant than those in cells only induced by PDGF-BB, with up-modulating rates at 51.8 and 58.7%, respectively (P < 0.001). We also observed that the effects of hTK1 gene delivery in inhibiting VSMCs proliferation, arresting cell cycling in G0/G1 phase and up-regulating the expression of p27Kip1 and p2lCip1 could be blocked by icatibant (Hoe 140), a specific bradykinin B2 receptor antagonist. Taken together, these results demonstrated that hTK1 overexpressed by recombinant adenovirus potently inhibits VSMCs proliferation that is required for neointimal hyperplasia and restenosis, and may activate p27Kip1 and p2lCip1 signaling pathways via bradykinin B2 receptor.

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Acknowledgments

This study was supported by grants from Fujian Natured Science Foundation (2008J0082), Key projects of Fujian provincial Science and technology (2008KJB-02), and The Key program from Bureau of Scientific Research of Fujian Medical University (09ZD005).

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Authors and Affiliations

  1. Fujian Hypertension Research Institution, The First Affilited Hospital of Fujian Medical University, Fuzhou, 350005, China

    Hui-Zhen Yu, Liang-di Xie, Peng-li Zhu, Chang-sheng Xu & Hua-jun Wang

  2. Fujian Provincial Hospital, Fujian Institute of Clinical Geriatrics, Provincial Clinical College, Fujian Medical University, Fuzhou, 35001, China

    Hui-Zhen Yu & Peng-li Zhu

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  1. Hui-Zhen Yu
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  2. Liang-di Xie
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Correspondence to Hui-Zhen Yu or Liang-di Xie.

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Yu, HZ., Xie, Ld., Zhu, Pl. et al. Human tissue kallikrein 1 gene delivery inhibits PDGF-BB-induced vascular smooth muscle cells proliferation and upregulates the expressions of p27Kip1 and p2lCip1 . Mol Cell Biochem 360, 363–371 (2012). https://doi.org/10.1007/s11010-011-1076-y

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  • DOI: https://doi.org/10.1007/s11010-011-1076-y

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