title: Loss of HSulf-1 expression enhances autocrine signaling mediated by amphiregulin in breast cancer.
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.
authors.affiliation: Department of Laboratory Medicine and Experimental Pathology, Mayo Clinic Cancer Center, Rochester, MN 55905, USA.
authors.forename: Keishi
authors.initials: K
authors.lastname: Narita
authors.forename: Jeremy
authors.initials: J
authors.lastname: Chien
authors.forename: Sally A
authors.initials: SA
authors.lastname: Mullany
authors.forename: Julie
authors.initials: J
authors.lastname: Staub
authors.forename: Xiang
authors.initials: X
authors.lastname: Qian
authors.forename: Wilma L
authors.initials: WL
authors.lastname: Lingle
authors.forename: Viji
authors.initials: V
authors.lastname: Shridhar
mesh_terms: D066258:Amphiregulin; D019898:Autocrine Communication; D001943:Breast Neoplasms / Q000378:metabolism* / Q000473:pathology; D018270:Carcinoma, Ductal, Breast / Q000378:metabolism / Q000473:pathology; D018275:Carcinoma, Lobular / Q000378:metabolism / Q000473:pathology; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D002470:Cell Survival; D056741:Cyclin D; D016213:Cyclins / Q000378:metabolism; D015536:Down-Regulation; D066255:EGF Family of Proteins; D004797:Enzyme-Linked Immunosorbent Assay; D004815:Epidermal Growth Factor / Q000037:antagonists & inhibitors / Q000276:immunology / Q000378:metabolism / Q000494:pharmacology; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D006023:Glycoproteins / Q000037:antagonists & inhibitors / Q000276:immunology / Q000378:metabolism*; D066257:Heparin-binding EGF-like Growth Factor; D006801:Humans; D017403:In Situ Hybridization; D036341:Intercellular Signaling Peptides and Proteins / Q000276:immunology / Q000378:metabolism*; D008875:Middle Aged; D009361:Neoplasm Invasiveness / Q000473:pathology; D009364:Neoplasm Recurrence, Local / Q000378:metabolism / Q000473:pathology; D034741:RNA, Small Interfering / Q000494:pharmacology; D012190:Retroviridae; D016196:S Phase; D015398:Signal Transduction; D015238:Sulfotransferases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D046888:Tissue Array Analysis
chemical_list: C587848:AREG protein, human; D066258:Amphiregulin; D056741:Cyclin D; D016213:Cyclins; D066255:EGF Family of Proteins; D006023:Glycoproteins; C587883:HBEGF protein, human; D066257:Heparin-binding EGF-like Growth Factor; D036341:Intercellular Signaling Peptides and Proteins; D034741:RNA, Small Interfering; D004815:Epidermal Growth Factor; C470791:SULF1 protein, human; D015238:Sulfotransferases