Growth factor regulation of hyaluronan synthesis and degradation in human dermal fibroblasts: importance of hyaluronan for the mitogenic response of PDGF-BB

Li, Lingli; Asteriou, Trias; Bernert, Berit; Heldin, Carl‐Henrik; Heldin, Paraskevi

doi:10.1042/bj20061757

Cited by 112 publications

(114 citation statements)

References 45 publications

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“…A number of growth factors have been reported to stimulate hyaluronan synthesis (20,22,23), often through mitogen-activated protein kinase (MAPK) pathways (23). In line with those reports, PD98059 (2 M), an inhibitor of MEK1, decreased hyaluronan synthesis by 39% (Fig.…”

Section: M␤cd Does Not Change the Size Of Hyaluronan Produced By Mcf-7supporting

confidence: 68%

Methyl-β-cyclodextrin Suppresses Hyaluronan Synthesis by Down-regulation of Hyaluronan Synthase 2 through Inhibition of Akt

Kultti

Kärnä

Rilla

et al. 2010

Journal of Biological Chemistry

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Hyaluronan synthases (HAS1-3) are integral plasma membrane proteins that synthesize hyaluronan, a cell surface and extracellular matrix polysaccharide necessary for many biological processes. It has been shown that HAS is partly localized in cholesterol-rich lipid rafts of MCF-7 cells, and cholesterol depletion with methyl-␤-cyclodextrin (M␤CD) suppresses hyaluronan secretion in smooth muscle cells. However, the mechanism by which cholesterol depletion inhibits hyaluronan production has remained unknown. We found that cholesterol depletion from MCF-7 cells by M␤CD inhibits synthesis but does not decrease the molecular mass of hyaluronan, suggesting no major influence on HAS stability in the membrane. The inhibition of hyaluronan synthesis was not due to the availability of HAS substrates UDP-GlcUA and UDP-GlcNAc. Instead, M␤CD specifically down-regulated the expression of HAS2 but not HAS1 or HAS3. Screening of signaling proteins after M␤CD treatment revealed that phosphorylation of Akt and its downstream target p70S6 kinase, both members of phosphoinositide 3-kinase-Akt pathway, were inhibited. Inhibitors of this pathway suppressed hyaluronan synthesis and HAS2 expression in MCF-7 cells, suggesting that the reduced hyaluronan synthesis by M␤CD is due to down-regulation of HAS2, mediated by the phosphoinositide 3-kinase-Akt-mTOR-p70S6K pathway.Cholesterol is an important membrane constituent of mammalian cells, plasma membrane in particular. It decreases the fluidity of the membrane and is a major component of the detergent-resistant membrane microdomains called lipid rafts (1). Increased serum cholesterol and cholesterol accumulation in atherosclerotic lesions have been long associated to cardiovascular disease (2) and atherosclerosis (3, 4), respectively. Abnormal cholesterol accumulation has been also found in some malignancies, such as prostate tumors (5, 6). Furthermore, recent studies suggest that inhibition of the rate-limiting enzyme in cholesterol synthesis (3-hydroxy-3-methyl-glutarylcoenzyme A reductase) by long-term statin therapy decreases the incidence of certain tumor types (7,8).Cellular cholesterol content can be modulated by the cholesterol depleting agent methyl-␤-cyclodextrin (M␤CD) 2 (9). M␤CD disrupts lipid rafts (10) and activates many signaling proteins, like epidermal growth factor receptor (11), extracellular signal-regulated kinase (ERK) (12, 13), p38 (11,14), Src (15), and ␤-catenin (16). On the other hand, M␤CD decreases the growth of MCF-7 breast cancer and A2780 ovarian cancer cells implanted subcutaneously in nude mice (17). In the MCF-7 model M␤CD was even more effective than doxorubicin (17).Hyaluronan, a large glycosaminoglycan mainly present in the extracellular matrix of vertebrates, is composed of repeating disaccharide units containing glucuronic acid (GlcUA) and N-acetylglucosamine (GlcNAc). It is produced at the plasma membrane by three hyaluronan synthases (HAS1-3). HASs use UDP-GlcUA and UDP-GlcNAc as substrates and extrude the forming hyaluronan chain to the extracellu...

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Section: M␤cd Does Not Change the Size Of Hyaluronan Produced By Mcf-7supporting

confidence: 68%

Methyl-β-cyclodextrin Suppresses Hyaluronan Synthesis by Down-regulation of Hyaluronan Synthase 2 through Inhibition of Akt

Kultti

Kärnä

Rilla

et al. 2010

Journal of Biological Chemistry

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“…HASs were also detected with anti-HAS2 IgG (5 g/ml; Ref. 28) and anti-HAS3 IgG (1 g/ml). For ubiquitination studies, the anti-ubiquitin P4D1 antibodies were used at a dilution of 1:200.…”

Section: Methodsmentioning

confidence: 99%

“…Determination of Synthesized Hyaluronan and HAS2 Expression by the Transfectants-Hyaluronan synthesized de novo by untransfected COS-1 cells or COS-1 cells transiently transfected with pcDNA3 vector alone (mock) as well as with tagged HAS2 or HAS3 constructs, was determined after different time periods by using a microtiter-based assay, as described previously (28). Similarly, the hyaluronan-synthesizing capacity of CHO-KI cells stably transfected with 6myc-HAS2, was also determined.…”

Section: Methodsmentioning

confidence: 99%

“…Antibodies-The following primary antibodies were used: rabbit antisera against HAS2 (CGR; (28)) and against HAS3 (Abcam); mouse monoclonal antibodies against Flag M2 (IgG 1 , Sigma-Aldrich), c-Myc (9E10 IgG 1 , Santa Cruz Biotechnology), influenza hemagglutinin A (HA) antiserum (anti-HA, Y-11; Santa Cruz Biotechnology) and to ubiquitin (P4D1 clone IgG 1 , Santa Cruz Biotechnology); goat anti-mouse and goat anti-rabbit horseradish peroxidase IgG (GE Healthcare).…”

Section: Methodsmentioning

confidence: 99%

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The Activity of Hyaluronan Synthase 2 Is Regulated by Dimerization and Ubiquitination

Karousou

Kamiryo

Skandalis

et al. 2010

Journal of Biological Chemistry

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114

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Hyaluronan is a component of the extracellular matrix, which affects tissue homeostasis. In this study, we investigated the regulatory mechanisms of one of the hyaluronan-synthesizing enzymes, HAS2. Ectopic expression of Flag-and 6myc-HAS2 in COS-1 cells followed by immunoprecipitation and immunoblotting revealed homodimers; after co-transfection with Flag-HAS3, also heterodimers were seen. Furthermore, the expressed HAS2 was ubiquitinated. We identified one acceptor site for ubiquitin on lysine residue 190. Mutation of this residue led to inactivation of the enzymatic activity of HAS2. Interestingly, K190R-mutated HAS2 formed dimers with wt HAS2 and quenched the activity of wt HAS2, thus demonstrating a functional role of the dimeric configuration.Hyaluronan is abundantly found in tissues throughout the body and has key roles in tissue organization and homeostasis (1). Abnormal accumulation of hyaluronan in tissues and serum is associated with the progression of many diseases, such as inflammatory diseases and cancer (2-5). The biosynthesis of hyaluronan is tightly regulated by growth factors and cytokines as well as other stimuli that promote wound healing, inflammation, or transformation. External regulatory signals, such as platelet-derived growth factor (PDGF)-BB, transforming growth factor (TGF)-␤, and phorbol 12-myristate 13-acetate (PMA), regulate both the size (6) and amount of the produced hyaluronan (7-14).The hyaluronan-synthesizing enzymes (HAS) 4 were cloned and characterized first in the bacterium Streptococcus pyogenes, and then in mammals, where three different HAS isoforms were characterized, HAS1, HAS2, and HAS3 (15, 16). Studies by us and other laboratories (6, 11) revealed that growth factormediated increase of hyaluronan synthesis is often due to induction of the HAS2 gene in fibroblasts, whereas external signals have been shown to predominantly regulate HAS1 and HAS3 transcripts in synoviocytes and keratinocytes, respectively (12,14,17). In addition to the regulation of HASs at the transcriptional level (6,11,13,14), there is evidence that the activity of HAS isoforms is regulated by phosphorylation by protein kinase C (PKC), protein kinase A (PKA), calcium-dependent protein kinase, and extracellular signal-regulated kinase (10, 18 -20). Prediction from their amino acid sequences suggests that HASs are multi-pass membrane proteins (15) that occur in the plasma membrane, but there are also evidence that much of the HAS enzymes reside at intracellular localizations including perinuclear membrane, endoplasmic reticulum (ER)-Golgi pathway, and endocytic vesicles (21-23).Ubiquitination of proteins affects their stability, activity, interaction with other proteins as well as subcellular localization and trafficking (24, 25). Modification of a protein with K48-linked poly-Ub chains can trigger the proteasomal degradation of the protein. The functional consequences of protein ubiquitination by K63-linked poly-Ub chains include activation of proteins or alteration of their trafficking. Protein modi...

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“…For example, increased hyaluronan production stimulates the phosphoinositide 3-kinase, MAP kinase and COX-2 pathways whereas antagonists of hyaluronan interactions suppress these pathways Misra et al, 2003;Misra et al, 2008). Interestingly, interactions between CD44 and receptor tyrosine kinases in normal cells may lead to very different outcomes depending on hyaluronan concentration (Li et al, 2007c;. Furthermore, since emmprin stimulates hyaluronan production (Marieb et al, 2004), it should come as no surprise that it also increases activation of ERBB2 , the activities of cell survival signaling pathways (Misra et al, 2003), and the ability to grow in an anchorage-independent manner (Marieb et al, 2004), in a hyaluronan-dependent fashion.…”

Section: Regulation Of Cell Survival Signaling Pathways By Hyaluronanmentioning

confidence: 99%

Hyaluronan, CD44 and Emmprin: Partners in cancer cell chemoresistance

Toole

Slomiany

2008

Drug Resistance Updates

160

140

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Hyaluronan is not only an important structural component of extracellular matrices but also interacts with cells during dynamic cell processes such as occur in cancer. Consequently, interactions of hyaluronan with tumor cells play important cooperative roles in various aspects of malignancy. Hyaluronan binds to several cell surface receptors, including CD44, thus leading to co-regulation of signaling pathways that are important in regulation of multidrug resistance to anticancer drugs, in particular anti-apoptotic pathways induced by activation of receptor tyrosine kinases. Emmprin, a cell surface glycoprotein of the Ig superfamily, stimulates hyaluronan production and downstream signaling consequences. Emmprin and CD44 also interact with various multidrug transporters of the ABC family and monocarboxylate transporters associated with resistance to cancer therapies. Moreover, hyaluronan-CD44 interactions are critical to these properties in the highly malignant, chemotherapy-resistant cancer stem-like cells. Perturbations of the hyaluronan-CD44 interaction at the plasma membrane by various antagonists result in attenuation of receptor tyrosine kinase and transporter activities and inhibition of tumor progression in vivo. These antagonists, especially small hyaluronan oligomers, may be useful in therapeutic strategies aimed at preventing tumor refractoriness or recurrence due to drug-resistant sub-populations within malignant cancers.

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Growth factor regulation of hyaluronan synthesis and degradation in human dermal fibroblasts: importance of hyaluronan for the mitogenic response of PDGF-BB

Cited by 112 publications

References 45 publications

Methyl-β-cyclodextrin Suppresses Hyaluronan Synthesis by Down-regulation of Hyaluronan Synthase 2 through Inhibition of Akt

Methyl-β-cyclodextrin Suppresses Hyaluronan Synthesis by Down-regulation of Hyaluronan Synthase 2 through Inhibition of Akt

The Activity of Hyaluronan Synthase 2 Is Regulated by Dimerization and Ubiquitination

Hyaluronan, CD44 and Emmprin: Partners in cancer cell chemoresistance

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