Estrogens and anti-estrogens have striking effects on growth of target tissues in vivo. In order to examine these effects in vitro, experimental methods for the rigorous assessment of rates of net DNA synthesis in tissue culture systems are required. The quantitation of DNA synthesis with inorganic [32P]orthophosphate is described. The limitations of more conventional [3H]thymidine labeling are illustrated in this hormonally responsive system. Estrogen administration increases the specific activity of acid-soluble phosphate and increases net DNA synthesis relative to controls in MCF-7 human breast cancer cells. Stimulation is most evident after 36 hr of hormone treatment. Conversely, tamoxifen produces substantial inhibition of net DNA synthesis after 36 hr of hormone treatment. Thymidine availability modulates the effect of estrogens and anti-estrogens on DNA synthesis and constitutes an independent experimental variable.
↵2 To whom requests for reprints should be addressed, at National Cancer Institute, NIH, Building 10, Room 6B02, Bethesda, Md. 20205.
- Received April 14, 1981.
- Accepted January 7, 1982.
- ©1982 American Association for Cancer Research.