title: A potential role for 6-sulfo sialyl Lewis X in metastasis of bladder urothelial carcinoma.
abstract: OBJECTIVE
  It is widely accepted that sialyl Lewis X (sLeX) and sialyl Lewis A (sLeA, also known as CA 19-9) glycans expressed on cancer cells function in E-selectin-mediated metastasis. Recently, it was reported that 6-sulfo sLeX glycans detected by the MECA-79 monoclonal antibody are expressed in roughly a quarter of gastric adenocarcinoma cases, and that these cases show a poorer prognosis than MECA-79-negative cases do. The present study was undertaken to assess expression of 6-sulfo sLeX glycans in bladder urothelial carcinoma and evaluate potential clinical implications.
  
  
  METHODS
  We analyzed 78 specimens representing bladder urothelial carcinoma, as well as 4 bladder urothelial carcinoma cell lines, by immunostaining with a battery of anticarbohydrate antibodies. We also undertook an E-selectin·IgM chimera binding assay to assess E-selectin binding to 6-sulfo sLeX expressed on bladder urothelial carcinoma cells and performed reverse transcription polymerase chain reaction and complementary DNA transfection to determine which N-acetylglucosamine-6-O-sulfotransferases function in 6-sulfo sLeX biosynthesis in those cells. Finally, we performed double-immunofluorescence staining for MECA-79 and either CD3 or CD8 to evaluate potential association between high endothelial venule (HEV)-like vessels and tumor-infiltrating T lymphocytes.
  
  
  RESULTS
  6-Sulfo sLeX glycans were expressed in ~20% of bladder urothelial carcinoma cases, particularly in plasmacytoid and micropapillary variants. Positive cells were also bound by E-selectin·IgM chimeras in a calcium-dependent manner. Transcripts encoding N-acetylglucosamine-6-O-sulfotransferase-2 were detected preferentially in HT-1197 bladder urothelial carcinoma cells expressing 6-sulfo sLeX, and transfection of the enzyme complementary DNA into HT-1376 cells, which do not express 6-sulfo sLeX glycans, resulted in cell surface expression of 6-sulfo sLeX. Furthermore, 6-sulfo sLeX glycans were expressed in HEV-like vessels induced in and around lymphocyte aggregates formed near carcinoma cell nests. These HEV-like vessel-associated tumor-infiltrating lymphocytes were composed primarily of CD3(+) T cells, with a fraction of CD8(+) cytotoxic T cells.
  
  
  CONCLUSIONS
  Our findings indicate that 6-sulfo sLeX glycans likely play 2 roles in bladder urothelial carcinoma progression: one in lymphocyte recruitment to enhance antitumor immune responses, and the other in E-selectin-mediated tumor cell adhesion to vascular endothelial cells, which is potentially associated with metastasis.
authors.affiliation: Division of Tumor Pathology, Department of Pathological Sciences, Faculty of Medical Sciences, University of Fukui, Eiheiji, Japan; Division of Urology, Department of Surgery, Faculty of Medical Sciences, University of Fukui, Eiheiji, Japan.
authors.forename: Minekatsu
authors.initials: M
authors.lastname: Taga
authors.affiliation: Division of Tumor Pathology, Department of Pathological Sciences, Faculty of Medical Sciences, University of Fukui, Eiheiji, Japan.
authors.forename: Hitomi
authors.initials: H
authors.lastname: Hoshino
authors.affiliation: Division of Tumor Pathology, Department of Pathological Sciences, Faculty of Medical Sciences, University of Fukui, Eiheiji, Japan.
authors.forename: Shulin
authors.initials: S
authors.lastname: Low
authors.affiliation: Division of Surgical Pathology, University of Fukui Hospital, Eiheiji, Japan.
authors.forename: Yoshiaki
authors.initials: Y
authors.lastname: Imamura
authors.affiliation: Division of Urology, Department of Surgery, Faculty of Medical Sciences, University of Fukui, Eiheiji, Japan.
authors.forename: Hideaki
authors.initials: H
authors.lastname: Ito
authors.affiliation: Division of Urology, Department of Surgery, Faculty of Medical Sciences, University of Fukui, Eiheiji, Japan.
authors.forename: Osamu
authors.initials: O
authors.lastname: Yokoyama
authors.affiliation: Division of Tumor Pathology, Department of Pathological Sciences, Faculty of Medical Sciences, University of Fukui, Eiheiji, Japan. Electronic address: motokoba@u-fukui.ac.jp.
authors.forename: Motohiro
authors.initials: M
authors.lastname: Kobayashi
mesh_terms: D000230:Adenocarcinoma / Q000235:genetics / Q000378:metabolism / Q000556:secondary*; D000368:Aged; D000369:Aged, 80 and over; D019040:E-Selectin / Q000235:genetics / Q000378:metabolism; D005260:Female; D005455:Fluorescent Antibody Technique; D005500:Follow-Up Studies; D006801:Humans; D007124:Immunoenzyme Techniques; D016256:Lewis X Antigen / Q000031:analogs & derivatives; D008297:Male; D008875:Middle Aged; D009367:Neoplasm Staging; D009844:Oligosaccharides / Q000235:genetics / Q000378:metabolism*; D011379:Prognosis; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D000080506:Sialyl Lewis X Antigen / Q000031:analogs & derivatives; D015238:Sulfotransferases / Q000235:genetics / Q000378:metabolism; D001749:Urinary Bladder Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D000097811:Carbohydrate Sulfotransferases
chemical_list: D019040:E-Selectin; D016256:Lewis X Antigen; D009844:Oligosaccharides; D012333:RNA, Messenger; D000080506:Sialyl Lewis X Antigen; D015238:Sulfotransferases; D000097811:Carbohydrate Sulfotransferases; C086949:6'-sulfated sialyl Lewis x; C546936:SELE protein, human
keywords: 6-Sulfo sialyl Lewis X; Antitumor immunity; Bladder urothelial carcinoma; E-selectin; High endothelial venule; Metastasis