Alpha-V-dependent outside-in signaling is required for the regulation of CD44 surface expression, MMP-2 secretion, and cell migration by osteopontin in human melanoma cells

Samanna, Venkatesababa; Hua, Wei; Ego-Osuala, D.; Chellaiah, Meenakshi A.

doi:10.1016/j.yexcr.2006.03.022

Cited by 62 publications

(61 citation statements)

References 67 publications

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“…After electrophoresis, the gels were washed and stained with Coomassie Brilliant Blue for 2-3 h as described previously (34). About 100 -150 g of protein was used for immunoblotting analysis.…”

Section: Methodsmentioning

confidence: 99%

Regulation of Sealing Ring Formation by L-plastin and Cortactin in Osteoclasts

Sadashivaiah

Chellaiah

2010

Journal of Biological Chemistry

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The aim of this study is to identify the exact mechanism(s) by which cytoskeletal structures are modulated during bone resorption. In this study, we have shown the possible role of different actin-binding and signaling proteins in the regulation of sealing ring formation. Our analyses have demonstrated a significant increase in cortactin and a corresponding decrease in L-plastin protein levels in osteoclasts subjected to bone resorption for 18 h in the presence of RANKL, M-CSF, and native bone particles. Time-dependent changes in the localization of L-plastin (in actin aggregates) and cortactin (in the sealing ring) suggest that these proteins may be involved in the initial and maturation phases of sealing ring formation, respectively. siRNA to cortactin inhibits this maturation process but not the formation of actin aggregates. Osteoclasts treated as above but with TNF-␣ demonstrated very similar effects as observed with RANKL. Osteoclasts treated with a neutralizing antibody to TNF-␣ displayed podosome-like structures in the entire subsurface and at the periphery of osteoclast. It is possible that TNF-␣ and RANKL-mediated signaling may play a role in the early phase of sealing ring configuration (i.e. either in the disassembly of podosomes or formation of actin aggregates). Furthermore, osteoclasts treated with alendronate or ␣v reduced the formation of the sealing ring but not actin aggregates. The present study demonstrates a novel mechanistic link between L-plastin and cortactin in sealing ring formation. These results suggest that actin aggregates formed by L-plastin independent of integrin signaling function as a core in assembling signaling molecules (integrin ␣v␤3, Src, cortactin, etc.) involved in the maturation process.Osteoclasts, the multinucleated and terminally differentiated giant cells, are involved in bone resorption. The adhesion of osteoclasts to the bone during bone resorption leads to the formation of the clear zone, an actin-rich ring-like adhesion zone circumscribing an area of bone resorption. Formation of a clear zone or sealing zone (also known as the sealing ring) has been considered to be a marker of osteoclast activation and is fundamental to the process of osteoclast bone resorption. Sealing ring formation is mediated by the dynamics of the actin cytoskeleton. Distinct pathways and signaling molecules have been shown to play roles in the organization of the sealing ring during bone resorption. Our previous observations in gelsolin null (Gsn Ϫ/Ϫ ) 2 osteoclasts demonstrated that deficiency of this protein blocks podosome assembly and motility. However, the cells still exhibit sealing ring and matrix resorption (1). Therefore, Gsn Ϫ/Ϫ osteoclasts are capable of resorbing bone, but the resorbed areas are small due to the absence of podosomes and the resulting hypomotile nature of osteoclasts (1). Observations in Gsn Ϫ/Ϫ osteoclasts also suggest that the organization of the sealing ring presumably reflect changes in the role of actinbinding proteins. Spatial configurations of actin fil...

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Section: Methodsmentioning

confidence: 99%

Regulation of Sealing Ring Formation by L-plastin and Cortactin in Osteoclasts

Sadashivaiah

Chellaiah

2010

Journal of Biological Chemistry

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“…The relevance and intensity in the changes of expression in genes linked with EMT in the training series led us to concentrate on validating some significantly up-regulated proteins (>2-fold change and FDR<0.2) in this category (N-cadherin, osteonectin/SPARC, and PKCa). Prompted by a literature search and the nearly significant results reported here, we also studied glypican 3, which has recently been described as a novel tumor marker for melanoma (17), and osteopontin, a protein belonging to this EMT group that has a crucial role in melanoma (18,19) and seems to be diagnostically useful in the detection of mesothelioma in serum obtained from peripheral blood (20). Figure 3 shows the results of the univariate analyses of selected proteins and representative immunohistochemistry for N-cadherin, osteonectin/SPARC, and osteopontin, which were the statistically significant markers.…”

Section: Resultsmentioning

confidence: 99%

A High-Throughput Study in Melanoma Identifies Epithelial-Mesenchymal Transition as a Major Determinant of Metastasis

Alonso¹,

Tracey²,

et al. 2007

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Metastatic disease is the primary cause of death in cutaneous malignant melanoma (CMM) patients. To understand the mechanisms of CMM metastasis and identify potential predictive markers, we analyzed gene-expression profiles of 34 vertical growth phase melanoma cases using cDNA microarrays. All patients had a minimum follow-up of 36 months. Twenty-one cases developed nodal metastatic disease and 13 did not. Comparison of gene expression profiling of metastatic and nonmetastatic melanoma cases identified 243 genes with a >2-fold differential expression ratio and a false discovery rate of <0.2 (206 up-regulated and 37 down-regulated). This set of genes included molecules involved in cell cycle and apoptosis regulation, epithelial-mesenchymal transition (EMT), signal transduction, nucleic acid binding and transcription, protein synthesis and degradation, metabolism, and a specific group of melanoma-and neural-related proteins. Validation of these expression data in an independent series of melanomas using tissue microarrays confirmed that the expression of a set of proteins included in the EMT group (N-cadherin, osteopontin, and SPARC/osteonectin) were significantly associated with metastasis development. Our results suggest that EMT-related genes contribute to the promotion of the metastatic phenotype in primary CMM by supporting specific adhesive, invasive, and migratory properties. These data give a better understanding of the biology of this aggressive tumor and may provide new prognostic and patient stratification markers in addition to potential therapeutic targets. [Cancer Res 2007;67(7):3450-60]

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“…In the present study we show upregulation of OPN and its receptors ␣ V ␤ 3 and CD44 in vivo, in adventitia of hypertensive calves, and in PH-Fibs in vitro. Since cooperation/functional interaction of CD44, ␣ V ␤ 3 , and MMPs have been shown to regulate fibroblast invasion (16,21,46), we tested the relationship between OPN expression and that of gelatinases MMP-2 and MMP-9. Our in vitro data demonstrate that the augmented proinvasive potential of PH-Fibs was regulated by OPN at least in part via MMP-2.…”

Section: Discussionmentioning

confidence: 99%

“…A well-recognized OPN receptor, ␣ V ␤ 3 integrin, has been shown to modulate several cellular functional modalities including activation of signal transduction, secretion of MMP-2, and cell motility/invasion (6,16,21,46,49,69). CD44 has been correlated with cell metastatic/invasion potential of cancer cells (10,18,27,62) and fibroblasts (30), and loss of CD44 expression in transformed cells resulted in impaired potential to form tumor nodules (19).…”

Section: Discussionmentioning

confidence: 99%

Osteopontin is an endogenous modulator of the constitutively activated phenotype of pulmonary adventitial fibroblasts in hypoxic pulmonary hypertension

Anwar

Frid

et al. 2012

American Journal of Physiology-Lung Cellular and Molecular Phys

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Osteopontin is an endogenous modulator of the constitutively activated phenotype of pulmonary adventitial fibroblasts in hypoxic pulmonary hypertension. Am J Physiol Lung Cell Mol Physiol 303: L1-L11, 2012. First published May 11, 2012; doi:10.1152/ajplung.00050.2012.-Increased cell proliferation and migration, of several cell types are key components of vascular remodeling observed in pulmonary hypertension (PH). Our previous data demonstrate that adventitial fibroblasts isolated from pulmonary arteries of chronically hypoxic hypertensive calves (termed PH-Fibs) exhibit a "constitutively activated" phenotype characterized by high proliferative and migratory potential. Osteopontin (OPN) has been shown to promote several cellular activities including growth and migration in cancer cells. We thus tested the hypothesis that elevated OPN expression confers the "activated" highly proproliferative and promigratory/invasive phenotype of PH-Fibs. Our results demonstrate that, both in vivo and ex vivo, PH-Fibs exhibited increased expression of OPN, as well as its cognate receptors, ␣ V␤3 and CD44, compared with control fibroblasts (CO-Fibs). Augmented OPN expression in PH-Fibs corresponded to their high proliferative, migratory, and invasive properties and constitutive activation of ERK1/2 and AKT signaling. OPN silencing via small interfering RNA or sequestering OPN production by specific antibodies led to decreased proliferation, migration, invasion, and attenuated ERK1/2, AKT phosphorylation in PH-Fibs. Furthermore, increasing OPN levels in CO-Fibs via recombinant OPN resulted in significant increases in their proliferative, migratory, and invasive capabilities to the levels resembling those of PH-Fibs. Thus our data suggest OPN as an essential contributor to the activated (highly proliferative, migratory, and proinvasive) phenotype of pulmonary adventitial fibroblasts in hypoxic PH. vascular remodeling; inflammation; matricellular protein; extracellular matrix protein; vascular proliferation ALL FORMS OF CHRONIC PULMONARY hypertension (PH) are characterized by structural and fibroproliferative changes in both small and large pulmonary arteries (PAs) through a process termed vascular remodeling. Remodeling can affect all layers of the vessel wall and involves changes in the phenotype and functional behavior of each of the principal cell types in the vessel wall. However, one of the most consistent findings in experimental models of PH, as well as in models of vascular injury and hypertension in the systemic circulation, is early and dramatic adventitial remodeling, characterized by fibroblast proliferation, migration, and differentiation (5, 33, 53-55, 58, 64, 65). Although the mechanisms involved in these responses remain obscure, the possibility that they are coordinated through interactions with surrounding extracellular matrix (ECM) proteins is one possible mechanism (55, 58).

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Alpha-V-dependent outside-in signaling is required for the regulation of CD44 surface expression, MMP-2 secretion, and cell migration by osteopontin in human melanoma cells

Cited by 62 publications

References 67 publications

Regulation of Sealing Ring Formation by L-plastin and Cortactin in Osteoclasts

Regulation of Sealing Ring Formation by L-plastin and Cortactin in Osteoclasts

A High-Throughput Study in Melanoma Identifies Epithelial-Mesenchymal Transition as a Major Determinant of Metastasis

Osteopontin is an endogenous modulator of the constitutively activated phenotype of pulmonary adventitial fibroblasts in hypoxic pulmonary hypertension

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