Induction of replicative DNA synthesis in quiescent human fibroblasts by DNA damaging agents.

Abstract
A marked induction of DNA replication was observed in confluent human diploid fibroblast cultures treated with low relatively nontoxic doses of UV radiation, N-methyl-N-nitrosourea (MNU), and N-acetoxy-2-acetylaminofluorene (AAAF). Isopycnic CsCl density gradient analysis of newly synthesized DNA labeled with BrdUrd indicated that most of the synthesis was semiconservative. The rate of semiconservative DNA synthesis was maximal 24 h after damage. This induction of DNA replication was greatest at .apprxeq. 3 J/m2 UV, 0.5 mM MNU, or 1.0 .mu.M AAAF; it was inhibited by hydroxyurea and aphidicolin and also occurred in repair-deficient xeroderma pigmentosum fibroblasts. Autoradiographic examination of both confluent cultures and serum-arrested cultures showed a large increase in the fraction of densely labeled (S phase) cells after UV treatment. These densely labeled cells retain the capacity for cell division and subsequent proliferation. Low doses of at least 3 different DNA damaging agents are thus capable of recruiting quiescent cells into a state of DNA replication similar to that observed in the normal cell cycle.

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