Ras, Protein Kinase Cζ, and IκB Kinases 1 and 2 Are Downstream Effectors of CD44 During the Activation of NF-κB by Hyaluronic Acid Fragments in T-24 Carcinoma Cells

Fitzgerald, Katherine A.; Bowie, Andrew; Skeffington, Barbara Sheehy; O’Neill, Luke A.J.

doi:10.4049/jimmunol.164.4.2053

Cited by 136 publications

(73 citation statements)

References 59 publications

(54 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…It has also been reported that gene expression of MMP-1, MMP-3, and MMP-9 is regulated by the activation of protein kinase C in chondrocytes (38,39). Protein kinase C was shown to be a downstream effector of CD44 during the activation of NF-B by HA fragments in T-24 carcinoma cells (40). Thus, HA 6 treatment seems to disturb the stabilization of cytoskeletal conformation accomplished by high molecular mass HA-CD44 interactions, consequently leading to the activation of transcription factors affecting catabolism.…”

Section: Discussionmentioning

confidence: 94%

Hyaluronan oligosaccharide–induced activation of transcription factors in bovine articular chondrocytes

Ohno¹,

Im²,

Knudson³

et al. 2005

Arthritis & Rheumatism

View full text Add to dashboard Cite

Objective. To document the activity profile of transcription factors following chondrocyte stimulation with hyaluronan (HA) hexasaccharides (HA 6 ) and to determine the expression of genes whose transcriptional activation is tightly associated with the transcription factors.Methods. Nuclear extracts from bovine articular chondrocytes treated with HA 6 were subjected to transcription factor protein-DNA array analysis. Electrophoretic mobility shift assay (EMSA) analyses were performed to confirm the results of protein-DNA array. The gene expressions of matrix metalloproteinase 3 (MMP-3), type II collagen, and cartilage oligomeric matrix protein (COMP) were examined by quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR), and protease activity was assessed by casein zymography.Results. In the protein-DNA array analysis, 12 transcription factors were up-regulated and 2 transcription factors were down-regulated in the chondrocytes treated with HA 6 . The transcription factors retinoic acid receptor (RAR), retinoid X receptor (RXR), and Sp-1 exhibited >2-fold increased activity by HA 6 treatment, as confirmed by EMSA. RT-PCR analysis showed that the expression levels of MMP-3, type II collagen, and COMP messenger RNA, which are tightly associated with the activation of RAR, RXR, or Sp-1, were upregulated by treatment with HA 6 . Addition of high molecular mass HA after HA 6 treatment resulted in abrogation of the MMP-3 induction. Conclusion.These results suggest that HA 6 increase the activity of multiple transcription factors in chondrocytes and signal the enhanced expression of key genes involved in cartilage-matrix remodeling and turnover. The data also demonstrate that high molecular mass HA has a potential to suppress the signaling activated by HA 6 .The glycosaminoglycan hyaluronan (HA) is a critical component of the extracellular matrix of articular cartilage. HA serves as a scaffold for the aggregation of cartilage proteoglycan (1) and, in this manner, forms a continuum of structure throughout the extracellular matrix. In addition, we have shown that HA facilitates the anchorage of these proteoglycan aggregates to the chondrocyte cell surface via its interaction with the membrane HA receptor CD44 (2-4). Thus, it is, in part, the interaction of chondrocytes directly with HA that constitutes the cell-matrix interaction that serves to regulate many aspects of chondrocyte metabolism (5). We have shown previously that one method of interference with the cell-matrix interactions of chondrocytes is through the use of excess small HA oligosaccharides, such as HA hexasaccharides (HA 6 ) (2,4,6). HA 6 compete with high molecular mass HA for CD44 binding; yet, they are too small in size to interfere with HA aggrecan or HA-link protein interactions (1,7,8).The consequences of this loss of cell-matrix interactions can be dramatic. In a previous study, we demonstrated that the addition of small HA oligosaccharides to human or bovine articular cartilage slices resulted in the apparent activation o...

show abstract

Section: Discussionmentioning

confidence: 94%

Hyaluronan oligosaccharide–induced activation of transcription factors in bovine articular chondrocytes

Ohno¹,

Im²,

Knudson³

et al. 2005

Arthritis & Rheumatism

View full text Add to dashboard Cite

show abstract

“…Following functional validation of this tet-regulated CD44 expression system, RNA was extracted from HA-stimulated MCF7F-B5 cells in the absence and presence of CD44 expression and the transcriptional targets of HA/CD44 signaling determined by microarray analysis. Prior studies have indicated that the fragment size of HA impacts on the signaling competency of CD44 (McKee et al, 1996;Fitzgerald et al, 2000). Accordingly, RNA samples were harvested from cells stimulated with HA of an approximate molecular weight of 220 kDa.…”

Section: Discussionmentioning

confidence: 99%

Cortactin underpins CD44-promoted invasion and adhesion of breast cancer cells to bone marrow endothelial cells

et al. 2006

View full text Add to dashboard Cite

Using a validated tetracycline (tet)-regulated MCF7-founder (MCF7F) expression system to modulate expression of CD44 standard form (CD44s), we report the functional importance of CD44s and that of a novel transcriptional target of hyaluronan (HA)/CD44s signaling, EMS1/cortactin, in underpinning breast cancer metastasis. In functional experiments, tet-regulated induction of CD44s potentiated the migration and invasion of MCF7F cells through HA-supplemented Matrigel. EMS1/cortactin was identified by expression profiling as a novel transcriptional target of HA/CD44 signaling, an association validated by quantitative PCR and immunoblotting experiments in a range of breast cancer cell lines. The mechanistic basis underpinning CD44-promoted transcription of EMS1/cortactin was shown to be dependent upon a NFjB mechanism, since pharmacological inhibition of IjKinase-2 or suppression of p65 Rel A expression attenuated CD44-induced increases in cortactin mRNA transcript levels. Overexpression of a c-myc tagged murine cortactin construct in the weakly invasive, CD44-deficient MCF7F and T47D cells potentiated their invasion. Furthermore, the functional importance of cortactin to CD44s-promoted metastasis was demonstrated by selective suppression of cortactin in CD44-expressing MCF7F-B5 and MDA-MB-231 breast cancer cells using RNAi, which was shown to result in attenuated CD44-promoted invasion and CD44-promoted adhesion to bone marrow endothelial cells (BMECs).

show abstract

“…Thus, the observation that HA 6 and not HA 4 have the potential to activate MMP-13 transcription is consistent with our hypothesis that the cell signaling in chondrocytes is initiated by the displacement of HMW-HA, resulting in the declustering of CD44 in the cell membrane and destabilization of the cortical cytoskeletal conformation. Cytoskeletal rearrangements are known to be involved in the induction of MMPs (65,66) and include activation of protein kinase C and NFB (67).…”

Section: Discussionmentioning

confidence: 99%

Hyaluronan Oligosaccharides Induce Matrix Metalloproteinase 13 via Transcriptional Activation of NFκB and p38 MAP Kinase in Articular Chondrocytes

Ohno

Knudson

et al. 2006

Journal of Biological Chemistry

114

View full text Add to dashboard Cite

Hyaluronan exerts a variety of biological effects on cells including changes in cell migration, proliferation, and matrix metabolism. However, the signaling pathways associated with the action of hyaluronan on cells have not been clearly defined. In some cells, signaling is induced by the loss of cell-hyaluronan interactions. The goal of this study was to use hyaluronan oligosaccharides as a molecular tool to explore the effects of changes in cell-hyaluronan interactions and determine the underlying molecular events that become activated. In this study, hyaluronan oligosaccharides induced the loss of extracellular matrix proteoglycan and collagen from cultured slices of normal adult human articular cartilage. This loss was coincident with an increased expression of matrix metalloproteinase (MMP)-13. MMP-13 expression was also induced in articular chondrocytes by hyaluronan (HA) hexasaccharides but not by HA tetrasaccharides nor high molecular weight hyaluronan. MMP-13 promoterreporter constructs in CD44-null COS-7 cells revealed that both CD44-dependent and CD44-independent events mediate the induction of MMP-13 by hyaluronan oligosaccharides. Electromobility gel shift assays demonstrated the activation of chondrocyte NFB by hyaluronan oligosaccharides. NFB activation was also documented in C-28/I2 immortalized human chondrocytes by luciferase promoter assays and phosphorylation of IKK-␣/␤. The link between activation of NFB and MMP-13 induction by HA oligosaccharides was further confirmed through the use of the NFB inhibitor helenalin. Inhibition of MAP kinases also demonstrated the involvement of p38 MAP kinase in the hyaluronan oligosaccharide induction of MMP-13. Our findings suggest that hyaluronan-CD44 interactions affect matrix metabolism via activation of NFB and p38 MAP kinase.In many tissues cell-matrix interactions serve to regulate cellular homeostasis (1, 2). Interference with these associations typically signal cells to initiate matrix repair, cell proliferation, or even cell migration. Such interactions are especially important in tissues such as articular cartilage, a tissue rich in extracellular matrix but with limited vascular access for systemic control of homeostasis. Chondrocytes are thus highly dependent on cell-matrix interactions as a primary means to "sense" changes in the extracellular environment (3). Most investigations in this area focus on cell signaling induced through the interaction of integrin receptors with collagens, fibronectin, laminin, matrilins, etc.(3-7). However, cell-matrix interactions involving hyaluronan (HA), 2 once thought to be predominately structural in nature, are now beginning to receive increasing attention as initiators of cell signaling.HA is a high molecular weight polysaccharide consisting of repeating disaccharide units glucuronic acid and N-acetylglucosamine (8). In cartilage and many other connective tissues, HA serves as a central filamentous scaffold to which proteoglycans such as aggrecan and link protein become bound (9). This complex matrix networ...

show abstract

Ras, Protein Kinase Cζ, and IκB Kinases 1 and 2 Are Downstream Effectors of CD44 During the Activation of NF-κB by Hyaluronic Acid Fragments in T-24 Carcinoma Cells

Cited by 136 publications

References 59 publications

Hyaluronan oligosaccharide–induced activation of transcription factors in bovine articular chondrocytes

Hyaluronan oligosaccharide–induced activation of transcription factors in bovine articular chondrocytes

Cortactin underpins CD44-promoted invasion and adhesion of breast cancer cells to bone marrow endothelial cells

Hyaluronan Oligosaccharides Induce Matrix Metalloproteinase 13 via Transcriptional Activation of NFκB and p38 MAP Kinase in Articular Chondrocytes

Contact Info

Product

Resources

About