Dual effects of the membrane-anchored MMP regulator RECK on chondrogenic differentiation of ATDC5 cells
ABSTRACT Extracellular matrix (ECM) undergoes continuous remodeling during mammalian development. Although involvement of matrix metalloproteinases (MMPs) in ECM degradation has been well documented, how this process is regulated to allow proper ECM accumulation remains unclear. We previously showed the involvement of a membrane-anchored MMP regulator, RECK (reversion-inducing cysteine-rich protein with Kazal motifs), in vascular development in mice. Here we report that Reck mRNA can be detected in developing cartilage in E13.5 approximately 16.5 mouse embryos and is progressively upregulated during differentiation of a chondrogenic cell line ATDC5 in vitro. In the early phase of ATDC5 differentiation, RECK expression stays low, multiple MMPs are upregulated, and there is ECM degradation at the sites of cellular condensation. In the later phase, RECK is upregulated inside the expanding cartilaginous nodules where type II collagen is accumulated while active ECM degradation persists along the rim of the nodules. Constitutive RECK expression suppressed initial cellular condensation, whereas RECK knockdown suppressed the later ECM accumulation in the cartilaginous nodules. These results suggest that RECK expression at the right place (in the core of the nodules) and at the right time (only in the later phase) is important for proper chondrogenesis and that RECK, together with MMPs, plays a crucial role in regulating dynamic processes of tissue morphogenesis.
- SourceAvailable from: Emmanuel João Nogueira Leal Silva
[Show abstract] [Hide abstract]
- "In contrast to normal epithelium, EMT reflects the loss of cellular adherence and contact growth, involving dysregulation of adhesion molecules and increased breakdown of the extracellular matrix by matrix metalloproteinases (MMPs) (Guarino et al., 2007; Neth et al., 2007). MMPs are a family of zinc-dependent proteinases that degrade most extracellular matrix components (Kondo et al., 2007). Approximately 20 members of the MMP family have been identified, sharing common structural and functional elements (Zucker and Vacirca, 2004). "
ABSTRACT: Intracellular calprotectin (S100A8/A9) functions in the control of the cell cycle checkpoint at G2/M. Dysregulation of S100A8/A9 appears to cause loss of the checkpoint, which frequently characterizes head and neck squamous cell carcinoma (HNSCC). In the present study, we analyzed carcinoma cells for other S100A8/A9-directed changes in malignant phenotype. Using a S100A8/A9-negative human carcinoma cell line (KB), transfection to express S100A8 and S100A9 caused selective down-regulation of MMP-2 and inhibited in vitro invasion and migration. Conversely, silencing of endogenous S100A8 and S100A9 expression in TR146 cells, a well-differentiated HNSCC cell line, increased MMP-2 activity and in vitro invasion and migration. When MMP-2 expression was silenced, cells appeared to assume a less malignant phenotype. To more closely model the architecture of cell growth in vivo, cells were grown in a 3D collagen substrate, which was compared to 2D. Growth on 3D substrates caused greater MMP-2 expression. Whereas hypermethylation of CpG islands occurs frequently in HNSCC, S100A8/A9-dependent regulation of MMP-2 could not be explained by modification of the upstream promoters of MMP2 or TIMP2. Collectively, these results suggest that intracellular S100A8/A9 contributes to the cancer cell phenotype by modulating MMP-2 expression and activity to regulate cell migration and mobility.The International Journal of Biochemistry & Cell Biology 09/2014; 55. DOI:10.1016/j.biocel.2014.09.007 · 4.24 Impact Factor
[Show abstract] [Hide abstract]
- "The RECK gene encodes a membrane-anchored glycoprotein capable of regulating several members of the MMP family (MMP2, MMP7, MMP9, and MT1-MMP)6789 and some other extracellular metalloproteinases such as CD13/aminopeptidase N and ADAM10810. RECK is expressed in multiple normal tissues, including developing vasculature, skeletal muscles, neuromuscular junctions, cartilage, fibroblasts, and neural precursor cells710111213. In cultured mouse embryo fibroblasts, RECK was found to be required for proper cell-substrate adhesion and persistent directional migration14. "
ABSTRACT: Expression of a mesenchymal phenotype is often associated with invasive/metastatic behaviors of carcinoma cells. Acquisition of a mesenchymal phenotype by a carcinoma cell is known as epithelial-mesenchymal transition (EMT). The membrane-anchored matrix metalloproteinase-regulator RECK is abundant in normal mesenchymal cells. In aggressive carcinomas, however, RECK expression is often downregulated. This apparent paradox prompted us to clarify the relationship between EMT and RECK. We found that TGFβ-induced E-cadherin downregulation, a hallmark of EMT, is accompanied by RECK-upregulation in a non-tumorigenic epithelial cell line (MCF10A). In contrast, the loss of E-cadherin expression is uncoupled from RECK-upregulation in carcinoma-derived cell lines (MCF7, MDA-MB-231, and A549). When RECK was artificially expressed in A549 cells, it showed little effect on EMT but elevated the level of integrin α5 and attenuated cell proliferation and migration. These findings implicate RECK in the regulation of proliferation and migration of normal epithelial cells after EMT and suggest how the uncoupling between EMT and RECK-upregulation impacts on the fates and behaviors of carcinoma cells.Scientific Reports 04/2014; 4:4568. DOI:10.1038/srep04568 · 5.58 Impact Factor
[Show abstract] [Hide abstract]
- "Finally, the underlying mesenchymal cells and the ECM appear to have decreased integrity compared to those underlying the DE of the control limb buds (Fig. 2D panels 12 and 11; supplementary material Fig. S2). Our previous study implicated Reck in chondrocyte differentiation in vitro (Kondo et al., 2007). To test whether reduced Reck expression in developing cartilage is responsible for the skeletal phenotype of low-Reck mice, we generated mutant mice (R1/-;Col2a1-Cre) in which Reck is inactivated selectively in the late-stage mesenchymal cells committed to the chondrocyte lineage. "
ABSTRACT: The membrane-anchored metalloproteinase-regulator RECK has been characterized as a tumor suppressor. Here we report that mice with reduced Reck-expression show limb abnormalities including right-dominant, forelimb-specific defects in postaxial skeletal elements. The forelimb buds of low-Reck mutants have an altered dorsal ectoderm with reduced Wnt7a and Igf2 expression, and hypotrophy in two signaling centers (i.e., ZPA and AER) that are essential for limb outgrowth and patterning. Reck is abundantly expressed in the anterior mesenchyme in normal limb buds; mesenchyme-specific Reck inactivation recapitulates the low-Reck phenotype; and some teratogens downregulate Reck in mesenchymal cells. Our findings illustrate a role for Reck in the mesenchymal-epithelial interactions essential for mammalian development.Biology Open 05/2012; 1(5):458-66. DOI:10.1242/bio.2012638 · 2.42 Impact Factor