Expression analysis of 0-series gangliosides in human cancer cell lines with monoclonal antibodies generated using knockout mice of ganglioside synthase genes.
Some gangliosides, sialic acid-containing glycosphingolipids, have been considered as tumor-associated antigens. GD1α or a GD1α synthase gene ST6GalNAc5 was reported to be involved in the metastasis of murine lymphomas or human breast cancers, respectively. But expression patterns of 0-series gangliosides GD1α and its precursor GM1b in human cancers have not yet been investigated mainly due to lack of specific antibodies. We established specific monoclonal antibodies (mAbs) reactive with GD1α or GM1b using gangliosides from brain tissues of GM3 synthase (St3gal5)-deficient mice as immunogens. We used GM2/GD2 synthase (B4galnt1)-deficient mice to immunize by liposomes embedded with GD1α or acidic glycolipid fractions from brain of St3gal5-deficient mice. Specificities of established mAbs as analyzed by enzyme-linked immunosorbent assay and thin-layer chromatography-immunostaining were very high among various gangliosides. Increased expression of GD1α and reduced GM1b in the St6galnac5 cDNA-transfected RAW117 cell line also substantiated the specificities of two mAbs. Then, we analyzed expression of GD1α and GM1b, and of relevant glycosyltransferase genes in various human cancer cell lines using generated anti-GD1α mAb 122 or anti-GM1b mAb MR155A-7. A few human cancer cell lines showed significant expression of these gangliosides with reasonable expression of relevant glycosyltransferase genes.
Department of Life and Biomedical Sciences, Chubu University Faculty of Life and Health Sciences, 1200 Matsumoto-cho, Kasugai 487-8501, Japan, Department of Biochemistry II, Nagoya University Graduate School of Medicine, 65 Tsurumai, Showa-ku, Nagoya 466-0065, Japan.
Robiul Hasan
RH
Bhuiyan
Department of Biochemistry II, Nagoya University Graduate School of Medicine, 65 Tsurumai, Showa-ku, Nagoya 466-0065, Japan.
Yuji
Y
Kondo
Department of Biochemistry II, Nagoya University Graduate School of Medicine, 65 Tsurumai, Showa-ku, Nagoya 466-0065, Japan.
Tokiaki
T
Yamaguchi
Department of Biochemistry II, Nagoya University Graduate School of Medicine, 65 Tsurumai, Showa-ku, Nagoya 466-0065, Japan.
Noriyo
N
Tokuda
Department of Life and Biomedical Sciences, Chubu University Faculty of Life and Health Sciences, 1200 Matsumoto-cho, Kasugai 487-8501, Japan.
Yuki
Y
Ohkawa
Department of Biochemistry II, Nagoya University Graduate School of Medicine, 65 Tsurumai, Showa-ku, Nagoya 466-0065, Japan.
Noboru
N
Hashimoto
Department of Biochemistry II, Nagoya University Graduate School of Medicine, 65 Tsurumai, Showa-ku, Nagoya 466-0065, Japan.
Yuhsuke
Y
Ohmi
Department of Biochemistry II, Nagoya University Graduate School of Medicine, 65 Tsurumai, Showa-ku, Nagoya 466-0065, Japan.
Yoshio
Y
Yamauchi
Department of Life and Biomedical Sciences, Chubu University Faculty of Life and Health Sciences, 1200 Matsumoto-cho, Kasugai 487-8501, Japan, Department of Biochemistry II, Nagoya University Graduate School of Medicine, 65 Tsurumai, Showa-ku, Nagoya 466-0065, Japan.
Keiko
K
Furukawa
Department of Biochemistry II, Nagoya University Graduate School of Medicine, 65 Tsurumai, Showa-ku, Nagoya 466-0065, Japan.
Tetsuya
T
Okajima
Department of Life and Biomedical Sciences, Chubu University Faculty of Life and Health Sciences, 1200 Matsumoto-cho, Kasugai 487-8501, Japan, Department of Biochemistry II, Nagoya University Graduate School of Medicine, 65 Tsurumai, Showa-ku, Nagoya 466-0065, Japan koichi@isc.chubu.ac.jp.
Koichi
K
Furukawa
D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics / Q000276:immunology; D000951:Antigens, Neoplasm / Q000235:genetics; D045744:Cell Line, Tumor; D005677:G(M1) Ganglioside / Q000031:analogs & derivatives* / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D005732:Gangliosides / Q000235:genetics / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic; D006028:Glycosphingolipids / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D006801:Humans; D051379:Mice; D018345:Mice, Knockout; D017350:N-Acetylgalactosaminyltransferases / Q000235:genetics* / Q000378:metabolism; D009362:Neoplasm Metastasis; D012799:Sialyltransferases / Q000235:genetics* / Q000378:metabolism
D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm; D005732:Gangliosides; D006028:Glycosphingolipids; C064020:ganglioside GD1alpha; D005677:G(M1) Ganglioside; C029624:ganglioside M1b; D017350:N-Acetylgalactosaminyltransferases; C000590840:beta-1,4-N-acetyl-galactosaminyl transferase 1, mouse; D012799:Sialyltransferases; C038525:CMP-N-acetylneuraminate-alpha-N-acetylgalactosaminide alpha-2,6-sialyltransferase
cancer; ganglioside; glycosyltransferase; knockout mice; monoclonal antibody