Immunological Analysis of Glycolipids on Cell Surfaces of Cultured Human Tumor Cell Lines1: Expression of Lactoneotetraosylceramide on Tumor Cell Surfaces

Abstract

Glycolipids of human cell lines of colonic adenocarcinoma (Colo 205 and BM 314), gastric tumor (AZ 521 and KATO-III), and lung tumor (A 549) were studied by the immunohistochemical fluorescence technique, flow cytometric analysis and immunostaining on thin layer chromatoplates with antibodies against gangliotriaosylceramide (Gg3Cer), gangliotetraosylceramide (Gg4Cer), fucogangliotetraosylceramide (Fuc-Gg4Cer), blood group B active lipid, globopentaosylceramide (Gb5Cer) and lactoneotetraosylceramide (nLc4Cer). Anti-nLc4Cer antibody was the only antibody which reacted with all the tumor cell lines used. The glycolipid fractions of each cell line separated by Iatrobeads column chromatography were immunostained with the six antibodies mentioned above on thin layer plates. The presence of nLc4Cer was detected in all cell lines. On the other hand, Gg4Cer was detected in gastric tumor cell lines, and Gg3Cer was detected in AZ 521. Based on these results, the tumor cell lines were analyzed by flow cytometry using anti-nLc4Cer antibody. About 70% of total cells in each cell line were separated as nLc4Cer-expressing cells. The present findings, together with the occurrence of nLc4Cer in ascitic fluids of cancer patients (Taki, T., Kojima, S., Seto, H., Yamada, H., & Matsumoto, M. (1984) J. Biochem. 96, 1257-1265), suggest that nLc4Cer may be a tumor-associated lipid.

Tyrosine kinases and gastric cancer

“…The results are summarized in Figure 2b, with the height of the bar depicting the intensity of the band, and hence the expression level. In this ®gure, four dierent human gastric cancer cell lines (two American gastric cancer cell lines, AGS (Barranco et al, 1983) and HR (Shimizu et al, 1991), and two Japanese gastric cancer cell lines, AZ521 (Taki et al, 1985) and KATO III (Sekiguchi et al, 1978)) are compared. Thirty tyrosine kinases were identi®ed from these cell lines including 10 non-receptor tyrosine kinases and 20 receptor tyrosine kinases.…”