Método de obtención de datos útiles para evaluar la respuesta al tratamiento con 5-fluorouracilo (5-FU)

Método in vitro de obtención de datos útiles para evaluar la respuesta al tratamiento con 5-fluorouracilo (5-FU). La invención describe un Método in vitro de obtención de datos útiles para evaluar la respuesta al tratamiento con 5-fluorouracilo (5-FU) sólo o en combinación con otros principios activos como el IFN alpha de pacientes con cáncer de colon y mama.Peer reviewedServicio Andaluz de Salud, Universidad de Granada, Consejo Superior de Investigaciones Científicas (España)B1 Patente sin examen previ

Modulation of myogenic differentiation in a human rhabdomyosarcoma cell line by a new derivative of 5-fluorouracil (QF-3602)

The in vitro study of mechanisms involved in drug-induced maturation has made it possible to use differentiation-based therapy in clinical practice. The goal of this new therapy is the development of specific agents to induce cancer cells to stop proliferating and express characteristics of normal cells. Recently, by structural modifications of 5-fluorouracil (5-FU), we synthesized a new pyrimidine acyclonucleoside-like compound, 1-{[3-(3-chloro-2-hydroxypropoxy)-1-methoxy]propyl}-5-fluorouracil (QF-3602), which showed in rhabdomyosarcoma cells a low toxicity and time-dependent growth inhibition. In this work, we compared the degree of myogenic differentiation of RD rhabdomyosarcoma (RMS) cells after treatment with QF-3602 and 5-FU. Scanning and transmission electron microscopy (SEM and TEM) and immunocytochemical analyses showed that QF-3602 induced the appearance of myofilaments along the myotube-like giant RD cells, an increase in fibronectin and a decrease in vimentin expression. In contrast, only minor changes were observed with 5-FU. Moreover, polymerase chain reaction (PCR) analyses showed that QF-3602 did not induce overexpression of the mdr 1 gene, a resistance mechanism that frequently appears in classical cytotoxic therapy in these tumors. Compounds obtained by structural modifications of 5-FU may be useful in differentiation therapy as a new approach to the treatment of RMS

Método de obtención de datos útiles para evaluar, predecir y/o pronosticar la respuesta al tratamiento con análogos de pirimidina

[EN]The invention relates to an in vitro method for obtaining data that can be used to predict and prognosticate response to treatment with pyrimidine analogues and, preferably, with 5-fluorouracil (5-FU). The invention also relates to a kit and to the uses thereof.[ES] Método in vitro de obtención de datos útiles para predecir y pronosticar la respuesta al tratamiento conanálogos de pirimidina, y más preferiblemente con 5-fluorouracilo (S-FU), kit Yusos.Peer reviewedServicio Andaluz de Salud, Universidad de Granada, Consejo Superior de Investigaciones CientíficasA1 Solicitud de patente con informe sobre el estado de la técnic

E phage gene transfection associated to chemotherapeutic agents increases apoptosis in lung and colon cancer cells

The limited ability of conventional therapies to achieve the long-term survival of metastatic lung and colon cancer patients suggests the need for new treatment options. In this respect, genes encoding cytotoxic proteins have been proposed as a new strategy to enhance the activity of drugs, and combined therapies involving such genes and classical antitumoral drugs have been studied intensively. The E gene from phiX174 encodes a membrane protein with a toxic domain that leads to a decrease in tumour cell growth rates. Therefore, in order to improve the anti-tumour effects of currently used chemotherapeutic drugs on cancer cells, we investigated the association of the E suicide gene with these antineoplastic drugs. The E gene has antitumoral effects in both lung and colon cancer cells. In addition, expression of this gene induces ultrastructural changes in lung cancer transfected cells (A-549), although the significance of these changes remains unknown. The effect of combined therapy (gene and cytotoxic therapy) enhances the inhibition of tumour cell proliferation in comparison to single treatments. Indeed, our in vitro results indicate that an experimental therapeutic strategy based on this combination of E gene therapy and cytotoxic drugs may result in a new treatment strategy for patients with advanced lung and colon cancer

Indirect immunofluorescence study on the cytoskeleton of normal (FG) and neoplastic (SGS/3A) cadmium treated fibroblasts

Cadmium is a heavy metal dangerous for the environment and for the human health, with well shown carcinogenic potentiality. Many studies have related the professional exposure of the Cadmium with human pulmonary, prostatic and renal tumors, and it would be a role also in the tumors of the liver, of the hemopoietic system, of the bladder and of the stomach. The aim of the current study is to examine in normal and neoplastic fibroblasts culture cells the modifications induced by the Cadmium at cellular level, in particular on the cytoskeleton, responsible not only of the intracellular transport of vesicles and cell organules, but also of their positioning and of the cellular integrity. Two fibroblastic cellular strains, normal (FG) and neoplastic (SGS/3A), have been incubated in a 5 microM Cadmium acetate added medium for 1, 8, 24 hours and studied by indirect immunofluorescence methods, particularly for the following proteins: Actin, Tubulin and Vimentin. The observations show in normal and neoplastic fibroblasts comparables modifications and anomalies of cytoskeletal shape. In both the cases the cellular morphology suffers drastic modifications, gradually evolving through intermediary shapes: from triangular and spindle-shaped in the normal fibroblasts to irregular, star-shaped, and globular in the neoplastic ones. The Cadmium action on the morphology of the normal and tumoral cells changes according to the time of incubation, producing structural alterations of the cytoskeletal. The modifications that start to be observable at the first hour of incubation are more evident after the eighth hour of exposure, reaching the maximum expression at the twenty-fourth hour, often with reduction of the total volume of the cells and loss of their ability to adhere to the substratum. Such modifications can be related to great alterations of the cellular membrane, producing the change of shape and the progressive partial separation from the substratum. The intermediary filaments seem to be less sensitive, from a morphological but not functional point of view, to the action of the Cadmium in comparison to the Actin and the microtubules that, on the contrary, seem to lose their proper morphological characteristics

Differentiation: an encouraging approach to anticancer therapy

Differentiation is a complex multistep process of cell specialization that begins with the installation of a genetic programme, named determination, specific for a cell lineage. Development of the differentiation programme includes the cell-type specific silencing of some genes and the expression of other genes, that regulate the biological functions associated with the cellular type and that distinguish the specialized cells. Terminal differentiation is the end stage of this process where the cells irreversibly lose their proliferative capacity and which represents a form of negative control of growing. Regulating molecules interact to produce the correct balance between cellular multiplication and differentiation during embryogenesis and the normal behaviour of an adult. Cancer is a process in which changes in regulating circuits are produced, such as proliferation control, the balance between cellular survival and programmed cellular death (apoptosis), the communication with neighbouring cells and with the extracellular matrix, angiogenesis, and finally, the migration of the tumoural cell, the invasion and metastasic dissemination. This process implies the progressive development of a more malign phenotype with an increase of genetic alterations involving genes at several levels of expression during long periods of time. These genetic changes uncouple the normal balance between multiplication and cellular differentiation with an increase in the rate of proliferating cells. Classic chemotherapeutical agents have been very important; nevertheless, as the mechanism of action of these drugs depends on the cytodestruction of the neoplastic cells, their beneficial effects are normally accompanied by a notable morbidity, cytotoxicity and multidrug resistance. The knowledge of the mechanisms involved in differentiation and malignant transformation has allowed the search of alternative routes for antitumoural therapy that does not imply cellular death. Differentiation therapy focuses on the development and use of specific agents designed to selectively attract the terminal differentiation process, making the elimination of tumoural cells feasible together with the establishment of normal cellular homeostasis