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Pyrimidine Metabolism in Tissue Culture Cells Derived from Rat Hepatomas

I. Suspension Cell Cultures Derived from the Novikoff Hepatoma¹

Culture conditions are described for several cell lines derived from the Novikoff rat hepatoma. Stability of the stem cell line is demonstrated by a constant generation time of 10 hr for more than 360 transfer generations, and by its continued ability to produce typical animal tumors after 4 years in culture.

As in the parent tumor, no catabolism of thymine or uracil could be demonstrated. Complete inhibition of growth of the cultures by 10^-3 M thymidine and 10^-3 M 5-fluoro-2'-deoxyuridine, and complete reversal of amethopterin and 5-fluoro-2'-deoxyuridine toxicity by 10^-5 M thymidine, provide indirect evidence for high levels of thymidine kinase in these cells. Thymidine toxicity was shown to be readily reversed by 10^-5 M deoxycytidine.

Two variant strains of cells are described, one of which apparently lacks thymidine kinase, since it is irreversibly inhibited by amethopterin in the presence of thymidine and is resistant to high levels of both thymidine and 5-fluoro-2'-deoxyuridine. The 2nd variant strain possesses thymidine kinase, but is resistant to high levels of thymidine.

¹ This work was supported in part by grant CA-00646 from the National Cancer Institute, NIH, USPHS. The data reported in this paper constitute a portion of a thesis submitted in partial fulfillment of the requirements for the Ph.D. degree by P. A. Morse, Jr., University of Wisconsin, 1963.

² Present address: Department of Microbiology, The University of Mississippi Medical Center, Jackson, Miss.

Received 6/29/64. Revised 12/11/64.

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