CEDIE   05498
CENTRO DE INVESTIGACIONES ENDOCRINOLOGICAS "DR. CESAR BERGADA"
Unidad Ejecutora - UE
congresos y reuniones científicas
Título:
“Characterization of tumors of the specific gonadal stroma in transgenic mice: biological and molecular aspects”.
Autor/es:
CHEMES HE
Lugar:
Clamart, Francia
Reunión:
Conferencia; Conferencia en convenio colaborativo INSERM-CONICET; 2007
Institución organizadora:
Endocrinologie et Génétique de la Reproduction et du Développement (INSERM)
Resumen:
Transgenic mice bearing a construct in which the expression of the SV40 oncogene is directed by the AMH promoter (AT mice) develop testicular tumors in adult life. We studied early steps of tumor development and characterized these tumors at different ages by means of histologic, morphometric immunohistochemical and molecular techniques. 1-3 month-old AT mice depicted multifocal Leydig cell hyperplasia. The Leydig cell volume was higher in some transgenic animals in comparison with littermate controls. Between 5½ and 7 months microscopic interstitial tumors developed in some AT mice that progressively developed and at 7-14 months of life formed large confluent tumors of high mitotic index. Tumor cells had the phenotypic characteristics of Leydig cells, or formed solid cord-like structures reminiscent of those seen in Sertoli cell tumors. Leydig cell hyperplasias and tumors expressed 3b-hydroxysteroid dehydrogenase (3b-HSD) and inhibin a-subunit. AMH expression was negative in Leydig cell-like regions and variable in Sertoli cell-like areas. The large T antigen of SV40 (SV40 T) and markers of cell proliferative activity (PCNA) were intensely positive in hyperplastic cells and tumors. Control mice of similar ages showed neither hyperplasia nor tumors, and SV40 T expression was always negative. In conclusion, many adult AT mice develop large testicular tumors that are preceded by interstitial hyperplasia and microtumors. The histologic and immunohistochemical phenotype of tumors in AT mice (Leydig and Sertoli cell differentiation, positive 3b-HSD and a Inhibin and variable AMH) suggests a mixed differentiation of somatic cells of the specialized gonadal stroma. To explore the cellular origin of these tumors we have recently studied 11-14 day old male embryos at the time of testicular differentiation using markers of Sertoli cells (Wilms Tumor antigen WT1, SOX 9 and AMH), Leydig cells (3b-HSD) and the transgene SV40T. As anticipated, AMH, WT1 and SV40T were expressed in all fetal Sertoli cells, and 3b-HSD was positive in all Leydig cells of control and transgenic embryos. Unexpectedly, SV40T was also focally expressed in some Leydig cells. These SV40T-positive Leydig cells were present at all fetal ages and in hyperplasias or tumors of postnatal transgenic mice suggesting that fetal Leydig cells modified by the expression of the transgene were the origin of postnatal hyperplasias and tumors. The interesting finding that SV40T, an oncogene driven by a promoter that is specifically active in early fetal Sertoli cells, also expresses in fetal Leydig cells and induces testicular tumors of mixed differentiation is compatible either with transdifferentiation from early Sertoli cells to Leydig cells or with the existence of a common cell precursor for Leydig and Sertoli cells in the specific stroma of the gonadal ridge. Current efforts in our laboratory have resulted in the successful isolation of RNA transcripts from areas in paraffin tissue sections that depicted specific differentiation patterns both in control and transgenic mice. These RNA transcripts can be retrotranscribed and amplified by PCR to study gene expression. Further efforts to characterize the origin of tumors as well as the mechanisms of hyperplasia and tumorigenesis in these animals are currently under way. Supported by Grants from CONICET (PIP 2565/00 and 5479), ANPCyT (PICT 9591) and INSERM-CONICET International Cooperation Program.