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Emergence of Differentiated Subclones from a Human Salivary Adenocarcinoma Cell Clone in Culture after Treatment with Sodium Butyrate¹

Second Department of Oral and Maxillofacial Surgery [M. A., H. Y., T. Y., Y. Y., M. S.], Central Laboratory for Clinical Investigation [Y. H.], and Department of Biochemistry [A. U.], Tokushima University School of Dentistry, 3 Kuramoto-cho, Tokushima 770, Japan

A human salivary gland adenocarcinoma cell line, which has ultrastructure and biological markers specific to the intercalated duct cells of human salivary glands, was cultured in 5 mM sodium butyrate for 12 days; then the cells were trypsinized, subcultured for an additional 16 days, and then transferred to growth medium without sodium butyrate. Morphological changes appeared about 1 wk after return to growth medium without sodium butyrate; cells being spindle or stellate in shape appeared in the treated cells, whereas the untreated cells were polygonal in shape. This morphologically altered phenotype persists after more than 14 mo of culture in growth medium without sodium butyrate. Of 40 subclones isolated, 2 clonal cell lines were established from the subculture and characterized. The other 38 subclones were accompanied by cell death during the subcultures. The clonal lines exhibited a phenotype similar to myoepithelial cells such as myosin, ß-chain of S-100 protein, myofilaments, and oxytocin receptor in addition to decreased tumorigenicity and anchorage-independent growth. These findings indicate that commitment to differentiation into myoepithelial cells and conversion from malignant to normal phenotype occur in a human salivary gland adenocarcinoma cell line following the sodium butyrate treatment.

¹ This investigation was supported in part by a grant-in-aid for cancer research (No. 59015074) from the Ministry of Education, Science, and Culture of Japan.

² To whom requests for reprints should be addressed.

Received 8/ 6/86. Revised 10/21/85. Accepted 10/24/86.

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