Cardiovascular Research Advance Access first published online on March 18, 2008
This version [Corrected Proof] published online on May 7, 2008
Cardiovascular Research, doi:10.1093/cvr/cvn079
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FK506 can activate transforming growth factor-β signalling in vascular smooth muscle cells and promote proliferation
1 Invasive Cardiology Unit, Clinica Pineta Grande, Castelvolturno, Italy
2 Department of Biochemistry and Medical Biotechnology, University of Naples Federico II, via Pansini, 5, 80131 Naples, Italy
3 Institute of Endocrinology and Experimental Oncology, Italian National Research Council (CNR), Naples, Italy
* Corresponding author. Tel: +39 0817463125; fax: +39 0817463205. E-mail address: romano{at}dbbm.unina.it
Aims: FK506-binding protein (FKBP) 12 is an inhibitor of transforming growth factor (TGF)-β type I receptors. Several lines of evidence support the view that TGF-β stimulates vascular smooth muscle cell (VSMC) proliferation and matrix accumulation. We investigated the effect of FK506, also known as tacrolimus, on cellular proliferation and on matrix protein production in human VSMCs.
Methods and results: We measured cell proliferation with flow cytometry using BrdU incorporation and fluorimetrically by measuring DNA concentration with Hoechst 33258. Western blot assay of whole-cell lysates was used to measure the levels of signalling proteins involved in proliferative pathways, in particular β-catenin, pErk, pAkt, pmTOR, and cyclin D1. Collagen synthesis was also investigated by Western blotting. The TGF-β signal was studied by both Western blotting and confocal microscopy. We used the SiRNA technique for FKBP12 gene silencing. Our results show that FK506 stimulates VSMC proliferation and collagen type I production. FK506 enhanced β-catenin levels and activated the extracellular signal-regulated kinase, Akt, and mammalian target of rapamycin kinase, which are important effectors of proliferation. Accordingly, cyclin D1 expression was increased. We also demonstrate that FK506 activates the TGF-β signal in VSMCs and that, through this mechanism, it stimulates cell proliferation.
Conclusion: FK506 can act as a growth factor for VSMCs.
KEYWORDS FK506; TGF-β; VSMC; Proliferation
Time for primary review: 32 days