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UPTAKE AND RELEASE OF 3 H‐MITOMYCIN C IN MOUSE P‐388 CELL CULTURES
Author(s) -
ørstavik Jon
Publication year - 1974
Publication title -
acta pathologica microbiologica scandinavica section b microbiology and immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.909
H-Index - 88
eISSN - 1600-0463
pISSN - 0365-5563
DOI - 10.1111/j.1699-0463.1974.tb02322.x
Subject(s) - mitomycin c , incubation , nucleic acid , thymidine , tritium , dna , intracellular , rna , nucleic acid metabolism , in vitro , chemistry , microbiology and biotechnology , biochemistry , biology , genetics , physics , gene , nuclear physics
Mouse P‐388 cells were grown in suspension culture and exposed to 3 H‐labelled mitomycin C for 1 hour. With concentrations from 0.5 to 50 μg of 3 H‐mitomycin C per ml cell culture, about 0.01 per cent of the radioactivity added was found in washed cells. Most of the intracellular antibiotic was present as cold acid soluble material, but significant amounts of tritium were found in the nucleic acid‐ and protein fractions. The ratio of 3 H per weight unit was of the same order of magnitude for DNA and RNA, but substantially lower for cell protein. Double labelling experiments using 14 C‐thymidine and 3 H‐mitomycin C were carried out to study the specific loss of antibiotic from the cells during post‐treatment incubation. Cold TCA soluble 3 H‐mitomycin C was rapidly released from the cells during the first hours after they had been transferred to non‐radioactive growth medium. Loss of 3 H‐radioactivity from the RNA‐ and protein fractions roughly paralleled the loss of 14 C radioactivity from the DNA fraction over a period of 3 days. The content of 3 H‐radioactivity in the DNA containing cell fraction increased during the first day of post‐treatment incubation, followed by a substantial drop during the second day. This drop was far more extensive than the corresponding loss of 14 C radioactivity. During the third day after exposure to mitomycin C the drops in 3 H and 14 C in the DNA fraction were not significantly different.