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Assay of mitogen-induced effects on cellular incorporation of precursors for scavenger, de novo, and net DNA synthesis. 1987

R B Dickson, and S Aitken, and M E Lippman

In summary we have presented data on [3H]dThd incorporation into DNA for a positive and negative growth modulator. These data clearly do not correspond to those based on net DNA synthesis (ortho[32P]phosphate incorporation into purified DNA) and previous knowledge of the effects of these hormones on cell number and cellular DNA accumulation. The paradox was resolved by directly analyzing effects of hormones on de novo pyrimidine biosynthesis. E2 stimulated both de novo and scavenger pathways in the first wave of DNA synthesis and only de novo in the second (and subsequent) waves. In contrast, tam progressively inhibited de novo pyrimidine biosynthesis. These hormonal effects on intracellulr pyrimidine pool sizes rendered [3H]dThd incorporation data by itself uninterpretable. [3H]dThd is a useful measure of DNA synthesis only if verified by independent measures of net DNA synthesis for the same time course and treatment conditions. In addition it may prove beneficial in various experimental systems to develop direct assays of de novo pyrimidine biosynthesis to assess mitogen effects. The experiments presented may also prove to be useful in evaluating the effects of mitogens and antimitogens on cells synchronized in the cell cycle by any of a variety of means.

UI MeSH Term Description Entries
D010710 Phosphates Inorganic salts of phosphoric acid. Inorganic Phosphate,Phosphates, Inorganic,Inorganic Phosphates,Orthophosphate,Phosphate,Phosphate, Inorganic
D010761 Phosphorus Radioisotopes Unstable isotopes of phosphorus that decay or disintegrate emitting radiation. P atoms with atomic weights 28-34 except 31 are radioactive phosphorus isotopes. Radioisotopes, Phosphorus
D011865 Radioisotope Dilution Technique Method for assessing flow through a system by injection of a known quantity of radionuclide into the system and monitoring its concentration over time at a specific point in the system. (From Dorland, 28th ed) Radioisotope Dilution Technic,Dilution Technic, Radioisotope,Dilution Technics, Radioisotope,Dilution Technique, Radioisotope,Dilution Techniques, Radioisotope,Radioisotope Dilution Technics,Radioisotope Dilution Techniques,Technic, Radioisotope Dilution,Technics, Radioisotope Dilution,Technique, Radioisotope Dilution,Techniques, Radioisotope Dilution
D002250 Carbon Radioisotopes Unstable isotopes of carbon that decay or disintegrate emitting radiation. C atoms with atomic weights 10, 11, and 14-16 are radioactive carbon isotopes. Radioisotopes, Carbon
D002460 Cell Line Established cell cultures that have the potential to propagate indefinitely. Cell Lines,Line, Cell,Lines, Cell
D004261 DNA Replication The process by which a DNA molecule is duplicated. Autonomous Replication,Replication, Autonomous,Autonomous Replications,DNA Replications,Replication, DNA,Replications, Autonomous,Replications, DNA
D006133 Growth Substances Signal molecules that are involved in the control of cell growth and differentiation. Mitogens, Endogenous,Endogenous Mitogens
D000085 Acetates Derivatives of ACETIC ACID. Included under this heading are a broad variety of acid forms, salts, esters, and amides that contain the carboxymethane structure. Acetate,Acetic Acid Esters,Acetic Acids,Acids, Acetic,Esters, Acetic Acid
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D013936 Thymidine A nucleoside in which THYMINE is linked to DEOXYRIBOSE. 2'-Deoxythymidine,Deoxythymidine,2' Deoxythymidine

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