epidermal-growth-factor and onapristone

The molecular mechanisms by which progestins and antiprogestins inhibit human breast cancer cell growth are essentially unknown. The mechanisms by which they mediate growth inhibition in human breast cancer cells and the expression of the putative autocrine/paracrine growth factors, epidermal growth factor and transforming growth factors alpha and beta-1 were studied under conditions in which progestin and antiprogestin inhibit the growth of T-47D human breast cancer cells in culture. Under the same conditions, the expression of genes such as c-myc, c-jun and c-fos, which are known to have important roles in growth and differentiation, has been measured. The results indicate that progestins and antiprogestins differentially regulate expression of these genes. The data are consistent with the conclusion that the mechanism of growth inhibition of these two agents differs, although an initial interaction with the progesterone receptor is a necessary first step in initiating the as yet ill-defined cascade of events leading to growth inhibition.

Topics: Breast Neoplasms; Cell Division; Epidermal Growth Factor; ErbB Receptors; Gene Expression Regulation, Neoplastic; Genes, fos; Genes, jun; Genes, myc; Gonanes; Humans; Medroxyprogesterone Acetate; Mifepristone; Neoplasm Proteins; Progestins; Transforming Growth Factors; Tumor Cells, Cultured

Depending on the tissue, progesterone is classified as a proliferative or a differentiative hormone. To explain this paradox, and to simplify analysis of its effects, we used a breast cancer cell line (T47D-YB) that constitutively expresses the B isoform of progesterone receptors. These cells are resistant to the proliferative effects of epidermal growth factor (EGF). Progesterone treatment accelerates T47D-YB cells through the first mitotic cell cycle, but arrests them in late G1 of the second cycle. This arrest is accompanied by decreased levels of cyclins D1, D3, and E, disappearance of cyclins A and B, and sequential induction of the cyclin-dependent kinase (cdk) inhibitors p21 and p27(Kip1). The retinoblastoma protein is hypophosphorylated and extensively down-regulated. The activity of the cell cycle-dependent protein kinase, cdk2, is regulated biphasically by progesterone: it increases initially, then decreases. This is consistent with the biphasic proliferative increase followed by arrest produced by one pulse of progesterone. A second treatment with progesterone cannot restart proliferation despite adequate levels of transcriptionally competent PR. Instead, a second progesterone dose delays the fall of p21 and enhances the rise of p27(Kip1), thereby intensifying the progesterone resistance in an autoinhibitory loop. However, during the progesterone-induced arrest, the cell cycling machinery is poised to restart. The first dose of progesterone increases the levels of EGF receptors and transiently sensitizes the cells to the proliferative effects of EGF. We conclude that progesterone is neither inherently proliferative nor antiproliferative, but that it is capable of stimulating or inhibiting cell growth depending on whether treatment is transient or continuous. We also suggest that the G1 arrest after progesterone treatment is accompanied by cellular changes that permit other, possibly tissue-specific, factors to influence the final proliferative or differentiative state.

Topics: Breast Neoplasms; CDC2-CDC28 Kinases; Cell Cycle Proteins; Cell Division; Cyclin A; Cyclin B; Cyclin-Dependent Kinase 2; Cyclin-Dependent Kinase Inhibitor p16; Cyclin-Dependent Kinase Inhibitor p21; Cyclin-Dependent Kinase Inhibitor p27; Cyclin-Dependent Kinases; Cyclins; Drug Synergism; Epidermal Growth Factor; ErbB Receptors; G1 Phase; Gene Expression Regulation; Gonanes; Hormone Antagonists; Humans; Microtubule-Associated Proteins; Mifepristone; Progesterone; Promegestone; Protein Serine-Threonine Kinases; Tumor Cells, Cultured; Tumor Suppressor Proteins

The ability of uterine stromal cells to respond to embryo attachment or an artificial decidual stimulus and the development and maintenance of decidual cells are primarily regulated by progesterone (P), yet few P-induced genes have been described. Previously, we reported that P stimulated the expression of heparin-binding epidermal growth factor-like growth factor (HB-EGF) messenger RNA (mRNA) in rat uterine stromal cells. We now present evidence that the effects of P on HB-EGF expression are blocked by treatment with ZK98299 (onapristone), indicating that they are receptor mediated. Additionally, animals treated with ZK98299 alone or together with P failed to respond to a decidual stimulus, suggesting that stromal cell sensitivity was inhibited. These results provide further evidence that HB-EGF plays a role in the development of stromal cell sensitivity to decidual stimuli. Expression of HB-EGF mRNA was also investigated in differentiating stromal cells after an artificial decidual stimulus. HB-EGF mRNA levels increased from 8-48 h after stimulation, the same interval in which stromal cells exhibit maximal mitotic activity. In situ hybridization revealed that stromal and decidual cells were the major sources of HB-EGF mRNA during this period. These results suggest that HB-EGF acts as an autocrine/paracrine factor in stimulating stromal cell proliferation after the induction of decidualization.

Topics: Animals; Decidua; Epidermal Growth Factor; Female; Gene Expression Regulation; Gonanes; Heparin; Progesterone; Rats; Rats, Sprague-Dawley; RNA, Messenger; Stromal Cells; Uterus