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Quantitative Determination of N-Glycolylneuraminic Acid Expression in Human Cancerous Tissues and Avian Lymphoma Cell Lines as a Tumor-associated Sialic Acid by Gas Chromatography-Mass Spectrometry¹

Department of Biochemistry, Faculty of Veterinary Medicine, Hokkaido University, Sapporo 060 [T. K., A. K., M. N.], and Department of Pathology, Research Institute for Microbial Disease, Osaka University, Osaka 565 [H. H., S. K.], Japan

N-Glycolylneuraminic acid (NeuGc) is distributed in most animals except humans and chickens. However, human and chicken cancerous tissues often synthesize this heterophilic sialic acid as a tumor-associated Hanganutziu-Deicher antigen [M. Naiki and H. Higashi, Adv. Exp. Med. Biol., 152: 445–456, 1982; H. Higashi et al., Cancer Res., 45: 3796–3802, 1985]. In this paper, NeuGc in humans cancerous tissues and chicken Marek's disease lymphoma cell lines was determined quantitatively with gas chromatography-mass spectrometry analysis using mass fragmentography. The detectable limit of NeuGc was 40 pg (0.12 pmol) in each injection using 5 ng of trideuteriomethyl ester trideuteriomethyl glycoside of the sialic acid as an internal standard sample when a pair of ions at m/e 386 and 389 was chosen for ion monitoring. NeuGc was detected in ganglioside-rich fractions of various human cancerous tissues from 5 of 8 patients examined but was not detected in glycosphingolipids of normal human tissues. The contents of NeuGc in these cancerous tissues ranged from 0.02 to 0.5% of the total sialic acid content. NeuGc was also detected in freeze-dried samples of 5 different cell lines from chicken Marek's disease lymphomas but was not detected in a cell line from chicken lymphoid leukosis lymphoma and normal chicken skeletal muscle tissue. The contents of NeuGc in the positive cell lines ranged from 0.03 to 0.11% of the total sialic acid content. These results indicate that NeuGc can be synthesized in both humans and chickens in some cancers.

¹ This work was supported by Grants-in-Aid for Scientific Research No. 63010023 and 63010027 from the Ministry of Education, Science and Culture of Japan.

² To whom requests for reprints should be addressed, at Department of Veterinary Science, National Institute for Health, Kamiosaki 2-10-35, Shinagawa-ku, Tokyo 141, Japan.

Received 9/ 5/90. Accepted 12/ 6/90.

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