No abstract

Estrogens play an important role in regulating the growth and differentiation of normal, premalignant and malignant cell types, especially breast epithelial cells, through interaction with two nuclear estrogen receptors (ER and ERIn this review, we present a brief overview of the actions of estrogens in the different steps of breast carcinogenesis, including cancer progression to metastasis, and of their clinical consequences in the prevention, prognosis and treatment of the disease. The requirement of estrogen receptors, mainly of the alpha subtype, in normal mammary gland differentiation and growth has been evidenced by estrogen receptor deficiency in animals. The promotion of breast cancer carcinogenesis by prolonged exposure to estrogens is well-documented and this has logically led to the use of antiestrogens as potentially chemopreventive agents. In breast cancer progression, however, the exact roles of estrogen receptors have been less well established but they may possibly be dual. Estrogens are mitogenic in ER-positive cells and antiestrogens are an efficient adjuvant therapy for these tumors. On the other hand, the fact that estrogens and their receptors protect against cancer cell invasiveness through distinct mechanisms in experimental models may explain why the presence of ER is associated with well-differentiated and less invasive tumors.2

Cathepsin-D is an independent marker of poor prognosis in human breast cancer. We previously showed that human wild-type cathepsin-D, as well as its mutated form devoid of proteolytic activity stably transfected in 3Y1-Ad12 cancer cells, stimulated tumor growth. To investigate the mechanisms by which human cathepsin-D and its catalytically-inactive counterpart promoted tumor growth in vivo, we quantified the expression of proliferating cell nuclear antigen, the number of blood vessels and of apoptotic cells in 3Y1-Ad12 tumor xenografts. We first verified that both human wild-type and mutated cathepsin-D were expressed at a high level in cathepsin-D xenografts, whereas no human cathepsin-D was detected in control xenografts. Our immunohistochemical studies then revealed that both wild-type cathepsin-D and catalytically-inactive cathepsin-D, increased proliferating cell nuclear antigen expression and tumor angiogenesis. Interestingly, wild-type cathepsin-D significantly inhibited tumor apoptosis, whereas catalytically-inactive cathepsin-D did not. We therefore propose that human cathepsin-D stimulates tumor growth by acting -directly or indirectly -as a mitogenic factor on both cancer and endothelial cells independently of its catalytic activity. Our overall results provide the first mechanistic evidences on the essential role of cathepsin-D at multiple tumor progression steps, affecting cell proliferation, angiogenesis and apoptosis.

With 2 figures in the text)We describe food selection by roe deer (Cuprealus capreolus) in relation to the food quality of the plants available (the concentrations of fibres, sugars, crude protein, and of phenolics and terpenes). Seven tame roe deer feeding in an oak-beech woodland edge used the majority (SO-94%) of the plant species available: they were therefore generalist feeders. However, they preferred only a small number of plant species in the different seasons: ivy (Hederu helix) in winter and autumn, dogwood (Cornus spp.) in summer, hornbeam (Curpinus befulus), hawthorn (Crutuegus spp.) and bluebells (Hyacinthoides non-scriptu) in spring. No preference or avoidance could be demonstrated for 74-85% of the available species, but 7-12 species were avoided according to the season. Though the avoided species were sometimes common, they never comprised an important part of the diet. In contrast, the preferred species made up only 4-13% of the available green matter but composed a large part of the diet (22-49%). Although generalists, the roe deer were therefore also highly selective feeders. The use of the different plant species was influenced by their availability, but to a different extent in the different seasons. Use of the plant species correlated negatively with the fibre content, as predicted from the digestive morphology of this concentrate selector. Preference/avoidance was related to the concentrations of soluble sugars, but not the crude protein content. Contrary to expectation, the roe deer preferred plants with a high concentration of protein-binding phenolics, suggesting that this deer has specific mechanisms for de-activating these compounds. Among the nutritional consequences of the feeding strategy of roe were a 17-35% higher soluble sugar content of the diet compared with the available green plant tissues, and an even greater increase in the phenolics. The crude protein contents and the dry matter digestibility of the diets tended to be slightly higher, and the fibre contents were lower, than those of the available green plant tissues.

In this study, we have analyzed the expression of TRIM24/TIF-1␣, a negative regulator of various transcription factors (including nuclear receptors and p53) at the genomic, mRNA, and protein levels in human breast tumors. In breast cancer biopsy specimens, TRIM24/TIF-1␣ mRNA levels (assessed by Real-Time Quantitative PCR or microarray expression profiling) were increased as compared to normal breast tissues. At the genomic level, array comparative genomic hybridization analysis showed that the TRIM24/TIF-1␣ locus (7q34) exhibited both gains and losses that correlated with mRNA levels. By re-analyzing a series of 238 tumors, high levels of TRIM24/TIF-1␣ mRNA significantly correlated with various markers of poor prognosis (such as the molecular subtype) and were associated with worse overall survival. By using a rabbit polyclonal antibody for immunochemistry, the TRIM24/TIF-1␣ protein was detected in nuclei of normal luminal epithelial breast cells, but not in myoepithelial cells. Tissue microarray analysis confirmed that its expression was increased in epithelial cells from normal to breast infiltrating duct carcinoma and correlated with worse overall survival. Altogether, this work is the first study that shows that overexpression of the TRIM24/TIF-1␣ gene in breast cancer is associated with poor prognosis and worse survival, and it suggests that this transcription coregulator may play a role in mammary carcinogenesis and represent a novel prognostic marker.

The effects of long‐term water stress on water and terpene contents of the foliage of Cupressus sempervirens were studied. A great deal of water was lost over 2 months before a remarkable stabilization. A strong decrease of all the classes of terpenes accompanied this dehydration. Mono‐ and sesquiterpene hydrocarbons and free terpenols were almost entirely metabolized, whereas esters and terpene glycosides rose slightly and remained at a constant level when the water content had stabilized. Although a significant part of the mono‐ and sesquiterpene hydrocarbons was emitted in the early stage of stress application, the major part was used by the plant in response to the drought conditions.

The biosynthesis of terpene hydrocarbons has been investigated in maritime pine (Pinus pinaster Ait.) seedling primary leaves under light and darkness and with different precursors. Impossible in darkness, the synthesis of monoterpenes (mainly α‐ and β‐pinene) is strongly activated by light. Only 14C‐carbonate and 14C‐acetate can be incorporated into monoterpenes. Activation by light is comparatively much more effective for seedling leaves previously cultivated under short days than in leaves from seedlings given long days. The spectral bands which are efficient for the synthesis of monoterpenes are located around 480 and 685 nm with 14C‐carbonate and 480 and 630 nm with l‐14C‐acetate. Furthermore, this light activation does not occur if leaf pieces instead of whole leaves are used for the incorporation experiments. When 2‐14C‐mevalonic acid and 1‐14C‐isopentenyl pyrosphosphate are applied as precursors, no radioactivity is recorded in monoterpene hydrocarbons even after light exposures. In contrast, sesquiterpene hydrocarbons (caryophyllene and humulene) are easily synthesized under light or darkness in intact or fragmented leaves from the different precursors of photosynthetic or exogenous origin. From these results the compartmentalization in the synthesis of C10 and C15 hydrocarbons appears clear. There is a metabolic cooperation between the photosynthetic tissues and the specific site of elaboration of C10 hydrocarbons, which site is located in the parts where the epithelial cells of resin ducts are functional. The synthesis of sesquiterpene hydrocarbons takes place in the whole leaf without activation by light.

Intact plastids from cell suspensions of Vitis vinifera L. cv. Muscat de Frontignan, free of detectable contamination by other particles as judged by the distribution of organelle-specific marker enzymes and by electron microscopy, exhibit geranyl-diphosphate synthase activity (EC 2.5.1.1). This synthase activity remains stable after tryptic digestion of unlysed organelles and is enhanced by plastid disruption. We conclude that the enzyme is located within the organelle. The possibility of an isopentenyl diphosphate/dimethylallyl diphosphate translocating system which would play a major role in the regulation of monoterpene metabolism is discussed.