Two soluble forms of β1,4-galactosyltransferase released from ovarian cancer cells and from COS-1 cells transfected with its cDNA

Daisuke Aoki, Eiko Saitoh, Yumi Matsumoto, Eiichiro Tominaga, Akira Hirasawa, Nobuyuki Susumu, Yasuhiro Udagawa, Shiro Nozawa

Research output: Contribution to journalArticle

Abstract

Employing RMG-1 cells derived from human ovarian adenocarcinoma and COS-1 cells transfected with cDNA for β1,4-galactosyltransferase, we first investigated the intracellular localization of this enzyme and secondly assayed separately the two types of soluble forms of it (designated as normal GaIT and GAT) in their culture supernatants. As results, the binding of anti-β1,4-galactosyltransferase monoclonal antibody was strongly positive at the Golgi area surrounding nuclei and the staining pattern was the same between RMG-1 cells and COS-1 transformants. These data demonstrated that the cells were producing β1,4-galactosyltransferase and had the ability to release it into the culture medium. As for the two soluble forms released from the cells, they were simultaneously detected in the culture medium by Western blotting and enzyme immunoassay using monoclonal antibodies, i.e., Mab8628 reactive to both GAT and normal-GaIT and Mab8513 specific for GAT, indicating that both were derived from RMG-1 and from the cDNA introduced into COS-1 cells. Since serum-GAT clinically reflects tumor growth more accurately or specifically than normal GaIT does, RMG-1 and COS-1 cells having cDNA for β1,4-galactosyltransferase would be promising as sources of these enzymes in order to investigate the differences in the behaviors of normal GaIT and GAT associated with ovarian malignant tumors.

Original languageEnglish
Pages (from-to)209-214
Number of pages6
JournalActa Histochemica et Cytochemica
Volume32
Issue number3
DOIs
Publication statusPublished - 1999 Jan 1

    Fingerprint

Keywords

  • Immunofluorography
  • RMG-1
  • Soluble form
  • Transfection
  • β1,4-galactosyltransferase

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Biochemistry
  • Physiology
  • Histology
  • Cell Biology

Cite this