Glycobiology, 2002, Vol. 12, No. 6 379-388
© 2002 Oxford University Press
Ectopic expression of a GlcNAc 6-O-sulfotransferase, GlcNAc6ST-2, in colonic mucinous adenocarcinoma
2 Department of Biochemistry, Sasaki Institute, 2-2, Kanda-Surugadai, Chiyoda-ku, Tokyo 101-0062, and Core Research for Evolutional Science and Technology (CREST) of the Japan Science and Technology Corporation, 2-3, Kanda-Surugadai, Chiyoda-ku, Tokyo 101-0062, Japan; and 3 Second Department of Pathology, Kagoshima University School of Medicine, 8-35–1, Sakuragaoka, Kagoshima 890-8520, Japan
The content of sulfated glycans having 6-O-sulfated GlcNAc residues alters in the course of colonic carcinogenesis. We previously characterized two GlcNAc 6-O-sulfotransferases (SulTs), SulT-a and -b, expressed in colonic normal tissues and adenocarcinomas [Seko et al. (2000) Glycobiology, 10, 919–929]. Levels of the enzymatic activities of SulT-a in normal colonic mucosa are higher than those in colonic adenocarcinomas, and the enzymatic activities of SulT-b are detected only in mucinous adenocarcinomas. To determine which GlcNAc 6-O-SulTs cloned so far correspond to SulT-a and -b, we expressed seven enzymes of a Gal/GalNAc/GlcNAc 6-O-SulT family in COS-7 cells and examined their substrate specificities in comparison with those of SulT-a and -b. GlcNAc6ST-2 (HEC-GlcNAc6ST, LSST, or GST-3) can recognize GlcNAcß1
3GalNAc
1-O-pNP as a good acceptor as well as other O-linked- and N-linked-type oligosaccharides, and its substrate specificity was similar to that of SulT-b. GlcNAc6ST-3(I-GlcNAc6ST or GST-4) preferred Galß13(GlcNAcß16)GalNAc1-O-pNP as an acceptor to the other oligosaccharides examined, and its specificity was similar to that of SulT-a. To confirm these correspondences, we further performed quantitative analyses of transcripts for GlcNAc6ST-2 and -3 genes by competitive RT-PCR. As a result, GlcNAc6ST-2 gene was expressed in almost all the mucinous adenocarcinomas examined and hardly expressed in normal colonic mucosa and nonmucinous adenocarcinoma. Expression levels of transcript for GlcNAc6ST-3 in normal mucosa were significantly higher than those in adenocarcinomas. From these results, it was indicated that GlcNAc6ST-2 corresponds to mucinous adenocarcinoma-specific SulT-b and that expression of GlcNAc6ST-3 is down-regulated in colonic adenocarcinomas.
1 To whom correspondence should be addressed; E-mail: yamashita{at}sasaki.or.jp
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