Immortalized Gastric Epithelial Cell Line GSM06 Synthesizes Hyaluronan under the Influence of Simian Virus 40 Large T-Antigen Expression.

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GSM06 is a cell line established from the stomach of transgenic mouse harboring a temperature-sensitive simian virus 40 (SV40) large T-antigen gene. 3H-labeled macromolecules produced by the cells incubated with [3H]glucosamine were characterized to examine whether or not GSM06 cells synthesize mucin (mucus glycoprotein). The GSM06 cells grew until a confluent monolayer formed at 33°C (the permissive temperature for SV40 large T-antigen expression), and the 3H-labeled macromolecules appeared in both cell extract and medium during culture for at least 1 week. Unexpectedly, almost all 3H-labeled macromolecules, which were excluded from a column of Sepharose CL-4B, were identified as hyaluronan by analyses using Sepharose CL-2B chromatography, cesium trifluoroacetate equilibrium centrifugation, treatment with dithiothreitol, and trypsin, hyaluronidase, and chondroitinase ABC digestion. At a nonpermissive temperature (39°C), GSM06 cells grew only slightly, but produced much more hyaluronan than at 33°C. The results indicate that GSM06 cells produce not mucin, but hyaluronan, and that the expression of large T-antigen may influence hyaluronan synthesis in GSM06 cells.
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