ABSTRACT
Replication timing of allelic gene pairs is strictly regulated according to expression, genome stability, and epigenetic changes, and tumorigenesis may be associated with changes in the allelic replication in various tumors. Our aim was to determine whether such alterations had a prognostic value in Ewing's family tumor (EFT) patients. The KIF14 and MDM4 / PI3KC 2ß and the centromeric satellite sequence of chromosomes 8 and 12 were used for replication timing assessments. Aneuploidy was assessed by enumerating the copy numbers of chromosomes 8 and 12. Replication timing and aneuploidy were detected cytogenetically using multicolors fluorescence in situ hybridization assay applied in 135 EFT. Patients with trisomy 8 presented an association with an asynchronous replication pattern (SD) of MDM4 / PI3KC 2ß genes ( p = 0.013). Trisomy 12 was associated with a synchronous pattern (DD) of KIF14 probe signals ( p = 0.04). The DD synchronous replication pattern of KIF14 showed a correlation with age ( p < 0.0001), and the SS synchronous replication pattern of the same locus showed a correlation with lung metastatic ( p = 0.012). The subgroup of patients presenting with multiplet signals of MDM4 / PI3KC 2ß showed an association with treatment response ( p = 0.045) and age ( p = 0.033). Replication pattern of KIF14 may, significantly, be associated with chromosomal instability as MDM4 / PI3KC 2ß may be a considerably new marker of poor treatment response in EFT patients.
ABSTRACT
Metastatic spread is the single-most powerful predictor of poor outcome in Ewing sarcoma (ES). Therefore targeting pathways that drive metastasis has tremendous potential to reduce the burden of disease in ES. We previously showed that activation of the ERBB4 tyrosine kinase suppresses anoikis, or detachment-induced cell death, and induces chemoresistance in ES cell lines in vitro. We now show that ERBB4 is transcriptionally overexpressed in ES cell lines derived from chemoresistant or metastatic ES tumours. ERBB4 activates the PI3K-Akt cascade and focal adhesion kinase (FAK), and both pathways contribute to ERBB4-mediated activation of the Rac1 GTPase in vitro and in vivo. ERBB4 augments tumour invasion and metastasis in vivo, and these effects are blocked by ERBB4 knockdown. ERBB4 expression correlates significantly with reduced disease-free survival, and increased expression is observed in metastatic compared to primary patient-matched ES biopsies. Our findings identify a novel ERBB4-PI3K-Akt-FAK-Rac1 pathway associated with aggressive disease in ES. These results predict that therapeutic targeting of ERBB4, alone or in combination with cytotoxic agents, may suppress the metastatic phenotype in ES.
Subject(s)
Bone Neoplasms/pathology , Bone and Bones/pathology , ErbB Receptors/genetics , ErbB Receptors/metabolism , Gene Expression Regulation, Neoplastic , Sarcoma, Ewing/pathology , Bone Neoplasms/genetics , Bone Neoplasms/metabolism , Bone and Bones/metabolism , Cell Line, Tumor , Cell Movement , Enzyme Activation , Humans , Neoplasm Metastasis/genetics , Neoplasm Metastasis/pathology , Phosphatidylinositol 3-Kinases/metabolism , Proto-Oncogene Proteins c-akt/metabolism , Receptor, ErbB-4 , Sarcoma, Ewing/genetics , Sarcoma, Ewing/metabolism , Signal Transduction , Up-Regulation , rac1 GTP-Binding Protein/metabolismABSTRACT
Introdução: A fusão EWS-ETS é um evento genético envolvido na patogênesis dos tumores de Ewing (TE); entretanto, a progressão dessa doença é devido à adição de outras alterações genéticas. Objetivo: Analisar o impacto prognóstico de regiões genéticas específicas em um série de pacientes com TE. Material e Métodos: Através de análises in silico usando o banco de dados Progenetix, os cromossomos 7, 8 e 12 e as regiões 1q e 16q foram identificadas como alterações genéticas mais recorrentes na progressão de TE, mas sem consistência quanto ao prognóstico. Lâminas de TMA, incluindo 132 pacientes com TE do Hopsital A.C. Camargo, foram construídas para análises de alterações no número de cópias gênicas (ANC) dos cromossomos 7, 8 12 e regiões 1q32.1 (KIF14/MDM4/PIK3C2B/DDX59) e 16q21 (CDH11), por hidridação por fluorescência in situ em núcleos interfásicos (iFISH). Essas análises foram feitas através da construção de sondas caseiras (BAC) (laboratório de citogenética molecular, Canadá) para as regiões 1q32.1 e 16q21 e através de sondas comerciais de centrômeros 7, 8 e 12. Ao final, a frequência de ANC foram correlacionadas com as características clínico-patológicas e sobrevida dos pacientes. Resultados: Análises de ANC mostraram ganhos em 1q32.1 em 15/121 (12.4%) pacientes. Anormalidades em 16q21 foram vistas em 24/115 (20.9%) pacientes: um grupo com 11/115 (9.6%) pacientes apresentaram deleções por hemizigose, e o outro, 13/115 (11.3%) pacientes apresentaram ganhos em 16q21. Aneussomias dos cromossomos 7, 8 e 12 foram vistas em 9.7%, 21.8% e 14.5%, respectivamente. Análises univariadas mostraram associação entre ganhos de 16q21 (p=0.029) ou 1q32.1 (p=0.005) e recorrência da doença em pacientes portadores de TE com doença localizada. As curvas de sobrevida global mostraram ganhos em 16q21 como prognóstico desfavorável (p=0.04) para doença localizada. ...
Subject(s)
Humans , Chromosome Aberrations , Bone Neoplasms , Prognosis , Sarcoma, EwingABSTRACT
UNLABELLED: We associated clinical-pathological features of 142 OSCC with the expression pattern of GLUT1 and GLUT3 in order to estimate their prognostic value. METHODS: Clinical-pathological features and overall survival data of 142 patients with Oral Squamous Cell Carcinoma (OSCC) were retrospectively reviewed from A.C.Camargo hospital records. A tissue microarray (TMA) was built for the immunohistochemical (IHC) analysis of GLUT 1 and GLUT 3. IHC results were evaluated according to the staining pattern and number of positive cells. RESULTS: GLUT 1 was over expressed in 50.3% of OSSC cases showing membrane staining pattern. However, nuclear expression was observed in 49.7% of the analyzed cases. GLUT 3 over expression was detected in 21.1% of OSCC cases. The pattern of GLUT 1 expression showed significant association with alcohol consumption (p = 0.004). Positive cell membrane GLUT 3 protein expression was associated with advanced clinic-staging of tumours (p = 0.005) as well as with vascular embolization (p = 0.005). Positive expression of GLUT 3 was associated with unfavorable free-disease survival (p = 0.021). CONCLUSION: GLUT1 and GLUT3 protein expression evaluated by immunohistochemistry are, significantly, indicators of poor prognosis outcome in oral squamous cell carcinoma, probably due to the enhanced glycolytic metabolism of more aggressive neoplastic cells.
