Characterization of Progesterone Receptor Subunits in Breast Cancer Cell Line, CAMA-1N

Abstract

In order to study whether cytosolic progesterone receptor (PRc) is involved in progesterone-induced proliferative arrest of breast cancer cells (CAMA-1N), submit composition of PRc in these cells was characterized. In the TSK G3000 high-performance liquid chromatography (HPLC) column, molybdate-stabilized PRc eluted as a single sharp peak immediately following the void volume. PRc from this peak had a sedimentation coefficient of 7.6 S and was eluted at 0.22 M salt from a linear gradient (0.1 to 0.4 M NaCl) in a DEAE column. When covalently labeled with [3H]R5020, analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and visualized byfluorography, PRc from crude cytosol exhibited two major protein bands with molecular weights of 105 ± 2.1 and 83 ± 3.2 kD, respectively. However, HPLC-purified PRc showed only one major band of 105 kD. These results show that PRc in CAMA-1N cells has steroid-binding components similar to other target tissues and provides a basis for further quantitative and qualitative analyses of PRc during cell proliferation and progesterone-induced growth arrest of CAMA cells.