A simple method to determine whole cell uptake of radiolabelled oestrogen and progesterone and their subcellular localization in breast cancer cell lines in monolayer culture.
Specific uptake of tritiated 17 beta-oestradiol and R5020, a synthetic progestin, in breast cancer cell lines ( MCF7 and T47D) growing in monolayer culture in multiwell plates has been shown. Binding characteristics, calculated by Scatchard analysis, indicate the presence of steroid receptors of similar affinities and capacities to those already obtained with broken cell preparations. Lysis of the cells by treatment with a hypotonic buffer reveals the subcellular localization of the receptors so the method can be used to study receptor dynamics such as nuclear translocation and processing. Cell growth can be measured by DNA determination directly in the multiwell plates. Thus, the method provides a convenient way of studying the effects of steroid hormones (or any antihormone or chemotherapeutic agent) on growth and receptor content of breast cancer cells in monolayer culture.
D001943:Breast Neoplasms / Q000378:metabolism*; D002455:Cell Division; D002460:Cell Line; D004247:DNA / Q000032:analysis; D004958:Estradiol / Q000378:metabolism*; D006801:Humans; D008722:Methods; D011374:Progesterone / Q000378:metabolism*; D011397:Promegestone / Q000378:metabolism; D013347:Subcellular Fractions / Q000378:metabolism; D013997:Time Factors
D011374:Progesterone; D004958:Estradiol; D004247:DNA; D011397:Promegestone