Cloning of cDNA sequences of a progestin-regulated mRNA from MCF7 human breast cancer cells.
A cDNA clone corresponding to an mRNA regulated by the progestin R5020, has been isolated by differential screening of a cDNA library from the MCF7 breast cancer cell line, which contains estrogen and progesterone receptors. This probe hybridized with a single species of poly A + RNA of 8-kb molecular weight as shown by Northern blot analysis and could also be used to total RNA preparation. This recombinant clone hybridized specifically to an mRNA coding for a 250,000 daltons protein when translated in vitro. This protein was identical to the 250 kDa progestin-regulated protein that we previously described (Biochem. Biophys. Res. Commun. 121, 421-427, 1984) as shown by immunoprecipitation with specific rabbit polyclonal antibodies. Dose-response curve and specificity studies show that the accumulation of the Pg8 mRNA and that of the 250-kDa protein was increased by 5 to 30-fold following progestin treatment and that this effect was mediated by the progesterone receptor. Time course of induction indicated that the accumulation of mRNA was rapid and preceded that of the protein. This is the first report on a cloned cDNA probe of progestin-regulated mRNA in human cell lines.
D001943:Breast Neoplasms; D002460:Cell Line; D003001:Cloning, Molecular; D004247:DNA / Q000032:analysis*; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006801:Humans; D007700:Kinetics; D008715:Methionine; D008970:Molecular Weight; D009363:Neoplasm Proteins / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics; D009649:Norpregnadienes / Q000494:pharmacology*; D009693:Nucleic Acid Hybridization; D011397:Promegestone / Q000494:pharmacology*; D012333:RNA, Messenger / Q000235:genetics*; D014158:Transcription, Genetic / Q000187:drug effects*
D009363:Neoplasm Proteins; D009649:Norpregnadienes; D012333:RNA, Messenger; D004247:DNA; D011397:Promegestone; D008715:Methionine