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Isolation and Chemical Characterization of Cell Surface Sialoglycopeptide Fractions during Progression of Rat Ascites Hepatoma AS-30D¹

The University of Texas Graduate School of Biomedical Sciences at Houston, and Department of Biochemistry, The University of Texas M. D. Anderson Hospital and Tumor Institute at Houston, Houston, Texas 77025

In order to ascertain whether chemical alterations in cell surface glycopeptides may be associated with tumor progression, we isolated sialogylcopeptide fractions from the surface of AS-30D rat ascites hepatoma cells before and after tumor progression. During the 70th to 90th transplantation generation of the AS-30D tumor, a morphological change and an increase in tumor virulence were observed. The tumor morphology progressed from a predominance of very large cell clusters to a predominance of single cells, doublets, triplets, and smaller clusters. The neuraminidase-labile, cell surface sialic acid of the AS-30D tumor decreased 27% after morphological progression of the tumor. Sialoglycopeptide fractions were prepared by papain digestion of cell suspensions obtained from the 11th to 14th, 49th to 55th, and 116th transplantation generations. These fractions were partially purified by trichloroacetic acid precipitation, dialysis, and gel filtration on Sephadex G-50. Compositional analysis and ion-exchange chromatographic behavior of the sialoglycopeptide fractions were similar except for a marked decrease in their glucose content associated with increased passage of the tumor.

¹ This research was supported by research grants from the American Cancer Society (P-451), the Paul and Mary Haas Foundation, and The Robert A. Welch Foundation (G-354).

² Recipient of a Rosalie B. Hite Predoctoral Fellowship in Cancer Research.

Received 8/18/71. Accepted 12/ 3/71.