Abstract
Heparan sulfate (HS) glycosaminoglycans are the oligosaccharide chains of heparan sulfate proteoglycans. The sulfation of HS glycosaminoglycan residues is required for its interaction with various heparin-binding growth factors to promote their biological activities to activate their high affinity receptor tyrosine kinases. We have identified HS glycosaminoglycan-6-O-endosulfatase HSulf-1 as a down-regulated gene in ovarian, breast, and several other cancer cell lines. Here we have shown that HSulf-1 inhibits autocrine activation of the EGFR-ERK (epidermal growth factor receptor-extracellular signal-regulated kinase) pathway induced by serum withdrawal in MDA-MB-468 breast cancer cells. Short hairpin RNA-mediated down-regulation of HSulf-1 in HSulf-1 clonal lines of MDA-MB-468 led to a significant increase in autocrine activation of ERK compared with vector only control. The autocrine signaling was also inhibited with neutralization antibodies against amphiregulin and HB-EGF, the heparin-binding growth factor family of the EGF superfamily. Furthermore, HSulf-1-mediated inhibition of autocrine signaling was associated with reduced cyclin D1 levels, leading to decreased S phase fraction and increased G(2)-M fraction, as well as increased cell death. Finally, evaluation of HSulf-1 expression levels in primary invasive breast tumors by RNA in situ hybridization indicated that HSulf-1 is down-regulated in the majority (60%) of tumors, with a predominant association with lobular histology. These data suggest a potential role of HSulf-1 down-regulation in mammary carcinogenesis.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
MeSH terms
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Amphiregulin
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Autocrine Communication*
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Breast Neoplasms / metabolism*
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Breast Neoplasms / pathology
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Carcinoma, Ductal, Breast / metabolism
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Carcinoma, Ductal, Breast / pathology
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Carcinoma, Lobular / metabolism
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Carcinoma, Lobular / pathology
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Cell Line, Tumor
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Cell Proliferation
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Cell Survival
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Cyclin D
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Cyclins / metabolism
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Down-Regulation
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EGF Family of Proteins
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Enzyme-Linked Immunosorbent Assay
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Epidermal Growth Factor / antagonists & inhibitors
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Epidermal Growth Factor / immunology
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Epidermal Growth Factor / metabolism
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Epidermal Growth Factor / pharmacology
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Female
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Gene Expression Regulation, Neoplastic
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Glycoproteins / antagonists & inhibitors
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Glycoproteins / immunology
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Glycoproteins / metabolism*
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Heparin-binding EGF-like Growth Factor
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Humans
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In Situ Hybridization
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Intercellular Signaling Peptides and Proteins / immunology
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Intercellular Signaling Peptides and Proteins / metabolism*
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Middle Aged
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Neoplasm Invasiveness / pathology
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Neoplasm Recurrence, Local / metabolism
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Neoplasm Recurrence, Local / pathology
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RNA, Small Interfering / pharmacology
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Retroviridae
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S Phase
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Signal Transduction
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Sulfotransferases / antagonists & inhibitors
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Sulfotransferases / genetics
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Sulfotransferases / metabolism*
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Tissue Array Analysis
Substances
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AREG protein, human
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Amphiregulin
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Cyclin D
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Cyclins
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EGF Family of Proteins
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Glycoproteins
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HBEGF protein, human
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Heparin-binding EGF-like Growth Factor
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Intercellular Signaling Peptides and Proteins
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RNA, Small Interfering
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Epidermal Growth Factor
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SULF1 protein, human
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Sulfotransferases