PTRF/cavin-1 is a protein that cooperates with caveolins (Cav-1, Cav-2 and Cav-3) to the morphogenesis of membranous caveolar domains in different cell types. In this study we investigated its expression in rhabdomyosarcoma (RMS), an aggressive soft-tissue myogenic tumor that mainly affects children and adolescents. Our expression studies performed by RT-PCR and Western blot analysis showed that PTRF/cavin-1 is mainly detectable, in concomitance with Cav-1 and Cav-2, during the in vitro growth of human RMS cell lines and two primary tumor cultures established from RMS developing in conditional mouse models. In contrast, the variable and limited degree of myodifferentiation in the different RMS lineages was characterized by down-regulation of PTRF/cavin-1, Cav-1 and Cav-2 and by the increment of Cav-3. Confocal microscopy analysis showed that PTRF/cavin-1 has a broad cellular distribution in RMS cells, ranging from nuclei and cytosol to plasma membrane. In addition, immunoprecipitation analysis showed that PTRF/cavin-1 and Cav-1 co-localize and physically interact at the plasma membrane, suggesting that this close association may play an important role in the progression of RMS.
Validation of PTRF/cavin-1 expression and plasma membrane association with caveolin-1 in human cell lines and mouse primary tumor cultures of rhabdomyosarcoma
FAGGI, Fiorella;Codenotti S.;MONTI, Eugenio;FANZANI, Alessandro
2013-01-01
Abstract
PTRF/cavin-1 is a protein that cooperates with caveolins (Cav-1, Cav-2 and Cav-3) to the morphogenesis of membranous caveolar domains in different cell types. In this study we investigated its expression in rhabdomyosarcoma (RMS), an aggressive soft-tissue myogenic tumor that mainly affects children and adolescents. Our expression studies performed by RT-PCR and Western blot analysis showed that PTRF/cavin-1 is mainly detectable, in concomitance with Cav-1 and Cav-2, during the in vitro growth of human RMS cell lines and two primary tumor cultures established from RMS developing in conditional mouse models. In contrast, the variable and limited degree of myodifferentiation in the different RMS lineages was characterized by down-regulation of PTRF/cavin-1, Cav-1 and Cav-2 and by the increment of Cav-3. Confocal microscopy analysis showed that PTRF/cavin-1 has a broad cellular distribution in RMS cells, ranging from nuclei and cytosol to plasma membrane. In addition, immunoprecipitation analysis showed that PTRF/cavin-1 and Cav-1 co-localize and physically interact at the plasma membrane, suggesting that this close association may play an important role in the progression of RMS.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
