title: Enhancive effects of Lewis y antigen on CD44-mediated adhesion and spreading of human ovarian cancer cell line RMG-I.
abstract: BACKGROUND
  This study aimed to investigate the molecular structural relationship between cell adhesive molecule CD44 and Lewis y antigen, and determine the effects of Lewis y antigen on CD44-mediated adhesion and spreading of ovarian cancer cell line RMG-I and the Lewis y antigen-overexpressed cell line RMG-I-H.
  
  
  METHODS
  The expression of CD44 in RMG-I and RMG-I-H cells before and after treatment of Lewis y monoclonal antibody was detected by immunocytochemistry; the expression of Lewis y antigen and CD44 was detected by Western Blot. The structural relationship between Lewis y antigen and CD44 was determined by immunoprecipitation and confocal laser scanning microscopy. The adhesion and spreading of RMG-I and RMG-I-H cells on hyaluronic acid (HA) were observed. The expression of CD44 mRNA in RMG-I and RMG-I-H cells was detected by real-time RT-PCR.
  
  
  RESULTS
  Immunocytochemistry revealed that the expression of CD44 was significantly higher in RMG-I-H cells than in RMG-I cells (P < 0.01), and its expression in both cell lines was significantly decreased after treatment of Lewis y monoclonal antibody (both P < 0.01). Western Blot confirmed that the content of CD44 in RMG-I-H cells was 1.46 times of that in RMG-I cells. The co-location of Lewis y antigen and CD44 was confirmed by co-immunoprecipitation. The co-expression of CD44 and Lewis y antigen in RMG-I-H cells was 2.24 times of that in RMG-I cells. The adhesion and spreading of RMG-I-H cells on HA were significantly enhanced as compared to those of RMG-I cells (P < 0.01), and this enhancement was inhibited by Lewis y monoclonal antibody (P < 0.01). The mRNA level of CD44 in both cell lines was similar (P > 0.05).
  
  
  CONCLUSIONS
  Lewis y antigen strengthens CD44-mediated adhesion and spreading of ovarian cancer cells.
authors.affiliation: Department of Obstetrics and Gynecology, Shengjing Hospital Affiliated to China Medical University, Shenyang, 110004, P R of China.
authors.forename: Lili
authors.initials: L
authors.lastname: Gao
authors.forename: Limei
authors.initials: L
authors.lastname: Yan
authors.forename: Bei
authors.initials: B
authors.lastname: Lin
authors.forename: Jian
authors.initials: J
authors.lastname: Gao
authors.forename: Xiuyun
authors.initials: X
authors.lastname: Liang
authors.forename: Yanyan
authors.initials: Y
authors.lastname: Wang
authors.forename: Juanjuan
authors.initials: J
authors.lastname: Liu
authors.forename: Shulan
authors.initials: S
authors.lastname: Zhang
authors.forename: Masao
authors.initials: M
authors.lastname: Iwamori
mesh_terms: D000911:Antibodies, Monoclonal; D002448:Cell Adhesion; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D005260:Female; D006801:Humans; D018960:Hyaluronan Receptors / Q000235:genetics / Q000378:metabolism*; D007983:Lewis Blood Group Antigens / Q000235:genetics / Q000378:metabolism*; D010051:Ovarian Neoplasms / Q000378:metabolism / Q000473:pathology*
chemical_list: D000911:Antibodies, Monoclonal; C497483:CD44 protein, human; D018960:Hyaluronan Receptors; D007983:Lewis Blood Group Antigens; C094162:Lewis Y antigen