Minocycline inhibits PDGF-BB-induced human aortic smooth muscle cell proliferation and migration by reversing miR-221- and -222-mediated RECK suppression
Introduction
Dysregulated proliferation and migration of arterial smooth muscle cells lead to vascular complications, including intimal hyperplasia. Among various growth factors, the Platelet-derived growth factor (PDGF) family exerts potent mitogenic and migratory effects on vascular smooth muscle cells, ultimately resulting in intimal hyperplasia [1,2]. There are four members in PDGF family, PDGF-A, –B, –C and –D, that dimerize and signal via PDGF receptors alpha and beta. Both PDGFRα and PDGFRβ are class III receptor tyrosine kinases, and form either homo or heterodimers depending upon the ligand [3]. Among various homo- or heterodimers of members of the PDGF family, the PDGF-BB homodimer appears to be highly potent in exerting mitogenic and migratory effects [3] via activation of multiple kinase-dependent signaling cascades, including activation of PI3K/AKT and mitogen activated protein kinases [4,5]. Furthermore, a significant role for oxidative stress and oxidative stress-sensitive NF-κB (nuclear factor kappa-light-chain-enhancer of activated B cells)-dependent signaling and its downstream effectors microRNA (miR)-221 and miR-222 have also been implicated. However, the potential targets of these miRs in the context of human arterial smooth muscle cell proliferation and migration have yet to be fully identified.
Minocycline (7-dimethylamino-6-dimethyl-6-deoxytetracycline) is a US Food and Drug Administration (FDA)-approved second-generation, semi-synthetic, orally active tetracycline antibiotic. However, its antibiotic-independent effects have also been described, including vasculoprotection. In fact, minocycline has been shown to inhibit vascular endothelial growth factor (VEGF)-induced aortic smooth muscle cell (SMC) migration by suppressing ERK (Extracellular Signal-Regulated Kinase)- and PI3K (Phosphoinositide 3-Kinase)/AKT (V-Akt Murine Thymoma Viral Oncogene Homolog)-dependent signaling and matrix metallopeptidase (MMP)-9 induction [6]. In fact, in a rat model of balloon injury, minocycline blunted neointima formation by inhibiting MMP expression and SMC migration [6,7]. Furthermore, minocycline reduced atherosclerotic plaque size in apolipoprotein E-deficient mice, a murine model of atherosclerosis, via a PARP1 (Poly[ADP-Ribose]Polymerase 1) and p27Kip1-dependent suppression of SMC proliferation [8]. These reports suggest that minocycline may have therapeutic potential in vascular proliferative diseases.
RECK (Reversion Inducing Cysteine Rich Protein with Kazal Motifs) is first described as a tumor suppressor gene [[9], [10], [11], [12]]. It is a glycosylphosphatidylinositol (GPI) anchored membrane protein known to inhibit secretion and activation of MMPs. It is expressed in various cell types, including adipocytes, vascular endothelial cells, smooth muscle cells, cardiomyocytes and cardiac fibroblasts [13]. We have previously reported that ectopic expression of RECK suppresses angiotensin II-mediated cardiac fibroblast migration in vitro [[14], [15], [16]].
Many types of cancer cells express low levels of RECK, possibly contributing to their malignant potential. Supporting this hypothesis, forced expression of RECK has been shown to suppress cancer cell migration and proliferation [17,18], suggesting that RECK inducers may have therapeutic potential in lowering tumor growth and metastasis. In an attempt to identify small molecular compounds that can induce Reck gene expression, Noda et al. have identified minocycline as a potent activator of RECK promoter activity using a reporter gene assay in RAS-transformed fibroblasts [19]. Here, we hypothesized that by upregulating RECK, minocycline inhibits PDGF-induced human ASMC (SMC) proliferation and migration. Our results show that PDGF-BB differentially regulates RECK and MMPs; suppresses RECK, but induces MMPs activation; resulting ultimately in increased SMC proliferation and migration. Further supporting our hypothesis, minocycline reversed PDGF-BB-induced RECK suppression and inhibited SMC proliferation and migration. To our knowledge, this is the first report demonstrating the role of RECK in minocycline-induced suppression of PDGF-BB-mediated SMC proliferation and migration. Our data suggest a possible therapeutic role for minocycline and other RECK inducers in vascular proliferative diseases.
Section snippets
Materials
Carrier-free recombinant human PDGF-BB protein was purchased from R&D Systems (#220-BB; Minneapolis, MN). The gp91 ds-tat (YGRKKRRQRRRCSTRIRRQL - NH2; #AS-63818) and its scrambled peptide control sgp91 ds-tat (YGRKKRRQRRRCLRITRQSR - NH2; #AS-63821) were purchased from AnaSpec (Fremont, CA, USA). One of the major sources of superoxide generation is NADPH oxidase (NOX). SMC express Nox2 (gp91phox) along with its cell membrane and cytosolic components [20]. Binding of gp91phox to the p47-p67-p40
Minocycline inhibits PDGF-BB-mediated PI3K/AKT- and ERK-dependent SMC migration and proliferation without affecting cell viability
Multiple signal transduction pathways are implicated in PDGF-induced SMC proliferation and migration, including AKT and ERK activation. Therefore, we investigated whether PDGF-BB induces SMC migration and proliferation via AKT and ERK activation, and whether minocycline blunts these responses. Indeed, PDGF-BB induced time dependent AKT activation, as evidenced by the increased levels of phosphor-AKT (Ser473) levels (Fig. 1A), an effect inhibited by the PI3K inhibitor Wortmannin, the AKT
Discussion
The molecular mechanisms underlying growth factor-induced smooth muscle cell migration and proliferation have yet to be fully investigated. Here we report for the first time that minocycline, a tetracycline antibiotic, inhibits PDGF-BB-mediated human aortic smooth muscle cell (SMC) migration and proliferation by reversing RECK suppression. RECK is an MMP inhibitor, and PDGF-BB suppressed its expression via AKT and ERK activation, ROS generation, NF-κB and AP-1 activation, and miR-221 and
Conflicts of interest
The authors declare no conflict of interest.
Acknowledgments
BC is a recipient of the Department of Veterans AffairsResearch Career Scientist award (#IK6BX004016-01), and is supported by the U.S. Department of Veterans Affairs, Office of Research and Development-Biomedical Laboratory Research and Development (ORD-BLRD) Service Award VA-I01-BX002255. Work in SM's lab is supported by NIH/NIAID R01AI119131. SS is supported by NIHR21-HL113705, PD by HL080682 and 1-U54 GM104940, and TY by American Heart Association15SDG25240022.
References (51)
- et al.
Platelet-derived growth factors and their receptors: structural and functional perspectives
Biochim. Biophys. Acta
(2013) - et al.
Minocycline reduces plaque size in diet induced atherosclerosis via p27(Kip1)
Atherosclerosis
(2011) - et al.
The membrane-anchored MMP inhibitor RECK is a key regulator of extracellular matrix integrity and angiogenesis
Cell
(2001) - et al.
A ras-related gene with transformation suppressor activity
Cell
(1989) - et al.
Angiotensin II stimulates cardiac fibroblast migration via the differential regulation of matrixins and RECK
J. Mol. Cell. Cardiol.
(2013) - et al.
Docosahexaenoic acid reverses angiotensin II-induced RECK suppression and cardiac fibroblast migration
Cell. Signal.
(2014) - et al.
CIKS (Act1 or TRAF3IP2) mediates high glucose-induced endothelial dysfunction
Cell. Signal.
(2013) - et al.
OxLDL induces endothelial dysfunction and death via TRAF3IP2: inhibition by HDL3 and AMPK activators
Free Radic. Biol. Med.
(2014) - et al.
CIKS (Act1 or TRAF3IP2) mediates Angiotensin-II-induced Interleukin-18 expression, and Nox2-dependent cardiomyocyte hypertrophy
J. Mol. Cell. Cardiol.
(2012) - et al.
Aldosterone-induced cardiomyocyte growth, and fibroblast migration and proliferation are mediated by TRAF3IP2
Cell. Signal.
(2015)
Induction of microRNA-221 by platelet-derived growth factor signaling is critical for modulation of vascular smooth muscle phenotype
J. Biol. Chem.
miR-221&222 regulate TRAIL resistance and enhance tumorigenicity through PTEN and TIMP3 downregulation
Cancer Cell
Minocycline hepatotoxicity: Clinical characterization and identification of HLA-B *35:02 as a risk factor
J. Hepatol.
Platelet-derived growth factor promotes smooth muscle migration and intimal thickening in a rat model of balloon angioplasty
J. Clin. Invest.
Inhibition of neointimal smooth muscle accumulation after angioplasty by an antibody to PDGF
Science
Protein kinase A antagonizes platelet-derived growth factor-induced signaling by mitogen-activated protein kinase in human arterial smooth muscle cells
Proc. Natl. Acad. Sci. U. S. A.
Different proliferative properties of smooth muscle cells of human arterial and venous bypass vessels: role of PDGF receptors, mitogen-activated protein kinase, and cyclin-dependent kinase inhibitors
Circulation
Minocycline exerts multiple inhibitory effects on vascular endothelial growth factor-induced smooth muscle cell migration: the role of ERK1/2, PI3K, and matrix metalloproteinases
Circ. Res.
Minocycline inhibits smooth muscle cell proliferation, migration and neointima formation after arterial injury
J. Cardiovasc. Pharmacol.
Detection of genes with a potential for suppressing the transformed phenotype associated with activated ras genes
Proc. Natl. Acad. Sci. U. S. A.
Characterization of a human MSX-2 cDNA and its fragment isolated as a transformation suppressor gene against v-Ki-ras oncogene
Oncogene
A gene atlas of the mouse and human protein-encoding transcriptomes
Proc. Natl. Acad. Sci. U. S. A.
Involvement of the SKP2-p27(KIP1) pathway in suppression of cancer cell proliferation by RECK
Oncogene
Regulation of matrix metalloproteinase-9 and inhibition of tumor invasion by the membrane-anchored glycoprotein RECK
Proc. Natl. Acad. Sci. U. S. A.
RECK: a novel suppressor of malignancy linking oncogenic signaling to extracellular matrix remodeling
Cancer Metastasis Rev.
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