RESUMEN
INTRODUCTION: Puberty is a period of increased susceptibility to factors that cause increased breast cancer risk in adulthood. Mammary end buds (EBs) that develop during puberty are believed to be the targets of breast cancer initiation. Whereas the role of estrogen (E) has been extensively studied in pubertal mammary gland development, the role of progesterone (P) during puberty is less defined. METHODS: Pubertal and prepubertal ovariectomized mice were treated with vehicle control (C), E, P, or E+P. Mammary glands from these mice were analyzed for changes in morphology, proliferation, and expression of the downstream targets amphiregulin (AREG) and receptor activator of NF-κB ligand (RANKL). RESULTS: P, acting specifically through the progesterone receptor, induced increases in mammary gland proliferation and EB formation that were associated with increased AREG expression in ducts and EBs. E, acting specifically through the estrogen receptor, produced similar responses also mediated by AREG. Blocking AREG action by treatment with an EGFR inhibitor completely abrogated the effect of P on EB formation and proliferation and significantly reduced proliferation within ducts. P also increased expression of RANKL, primarily in ducts. Treatment with RANK-Fc, an inhibitor of RANKL, reduced P-dependent proliferation in ducts and to a lesser extent in EB, but did not cause EB regression. CONCLUSIONS: These results demonstrate a novel P-specific effect through AREG to cause EB formation and proliferation in the developing mammary gland both before and during puberty. Thus, hormones and/or factors in addition to E that upregulate AREG can promote mammary gland development and have the potential to affect breast cancer risk associated with pubertal mammary gland development.
Asunto(s)
Anfirregulina/biosíntesis , Estrógenos/metabolismo , Glándulas Mamarias Animales/crecimiento & desarrollo , Progesterona/metabolismo , Anfirregulina/metabolismo , Animales , Proliferación Celular/efectos de los fármacos , Estrógenos/administración & dosificación , Femenino , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Humanos , Glándulas Mamarias Animales/efectos de los fármacos , Ratones , Ovariectomía , Progesterona/administración & dosificación , Pubertad/efectos de los fármacos , Pubertad/metabolismo , Ligando RANK/biosíntesis , Factores de RiesgoRESUMEN
Risk-sensitive foraging theory (RSFT) was developed to explain a choice between a variable (risk-prone) or constant (risk-averse) option. In the RSFT literature, qualitative shifts in risk-sensitivity have been explained by fluctuations in daily caloric energy budget (DEB). The DEB rule describes foragers' choices as being based on fitness and rate of gain. If the DEB rule is correct, rewards that differ in caloric returns should cause differences in foragers' sensitivity to risk. However, few studies have explored the influence of reward quality on risk-sensitivity in mammals. The present study was designed to examine the effects of reward quality on risk-sensitivity when reward magnitude, delay to reward, body mass, and response effort were controlled. Results from the current study demonstrated that subjects rewarded with a high calorie reward (i.e., sugar) made significantly fewer choices for a variable option than subjects rewarded with a lower calorie reward (i.e., grain). These results are consistent with the predictions of the DEB rule, and add to the RSFT literature where reward quality was manipulated by describing difference in risk-sensitivity in mammals. Suggestions for future research include an examination of risk-sensitivity where flavor and caloric return are manipulated.
Asunto(s)
Ingestión de Energía/fisiología , Conducta Alimentaria/psicología , Ratas Sprague-Dawley/psicología , Recompensa , Animales , Masculino , Ratas , Factores de RiesgoRESUMEN
Epithelial-stromal cell interactions are important for normal development and function of the mouse mammary gland. The steroid hormone estrogen is required for epithelial cell proliferation and ductal development in vivo. Recent studies of estrogen receptor alpha knockout mice indicate that estrogen-induced proliferation is dependent upon the presence of estrogen receptor in mammary stromal cells, but not in epithelial cells. The purpose of the present study was to identify the underlying mechanism of estrogen-dependent stroma-derived effects on mammary epithelium. We have developed a minimally supplemented serum-free medium, collagen gel primary mammary coculture system to address the issue of stroma-derived, estrogen-dependent effects on epithelial cell proliferation. Conditioned medium from mammary fibroblasts or coculture with mammary fibroblasts caused increased epithelial cell proliferation and produced tubular/ductal morphology. Hepatocyte growth factor (HGF) was identified as the mediator of this effect, as the proliferative activity in fibroblast-conditioned medium was completely abolished by neutralizing antibody to HGF, whereas neutralizing antibodies to either epidermal growth factor or IGF-I had no effect. Treatment of mammary fibroblasts with estrogen increased the production of HGF. From these results we conclude that estrogen may indirectly mediate mammary epithelial cell proliferation via the regulation of HGF in mammary stromal cells and that HGF plays a crucial role in estrogen-induced proliferation in vivo.
Asunto(s)
División Celular/efectos de los fármacos , Medio de Cultivo Libre de Suero , Estrógenos/farmacología , Factor de Crecimiento de Hepatocito/fisiología , Glándulas Mamarias Animales/citología , Células del Estroma/química , Animales , Anticuerpos/farmacología , Células Cultivadas , Técnicas de Cocultivo , Colágeno , Medios de Cultivo Condicionados , Células Epiteliales/citología , Femenino , Fibroblastos/fisiología , Factor de Crecimiento de Hepatocito/análisis , Factor de Crecimiento de Hepatocito/inmunología , Humanos , Inmunohistoquímica , Factor I del Crecimiento Similar a la Insulina/farmacología , Glándulas Mamarias Animales/química , Ratones , Ratones Endogámicos BALB C , Receptores de Estrógenos/análisis , Proteínas Recombinantes/análisisRESUMEN
The steroid hormones, estrogen and progesterone, are required for mammary epithelial cell proliferation and alveolar morphogenesis in vivo. We have developed a minimally supplemented, serum-free medium, collagen gel primary mammary culture system to determine the mechanism of progestin-induced proliferation and alveolar morphogenesis. In epithelial cells cultured alone, treatment with progestin (R5020) alone produced a lumen within the epithelial organoids, but did not stimulate epithelial cell proliferation. The formation of lumens was associated with increased apoptosis, targeted within the organoids. We have previously reported that in our culture system hepatocyte growth factor (HGF) increases epithelial cell proliferation and induces a tubulo-ductal morphological response. In the present report we show that treatment with HGF and progestin (R5020) further increases epithelial proliferation above that with HGF alone and also produces an alveolar-like morphology similar to that observed in vivo in response to progestin treatment. To the best of our knowledge this is the first in vitro demonstration of both progestin-induced proliferation and alveolar-like morphogenesis of normal nonpregnant mouse mammary epithelial cells in vitro. These results suggest that HGF may play a crucial role in progestin-induced proliferation and morphogenesis in vivo.