title: Expression and activity of vascular endothelial growth factor and metalloproteinases in alveolar and embryonal rhabdomyosarcoma cell lines.
abstract: Rhabdomyosarcoma (RMS) is a malignant tumour of skeletal muscle origin which includes two major histological subtypes: alveolar rhabdomyosarcoma (ARMS), the more aggressive, and embryonal rhabdomyosarcoma (ERMS). In order to establish whether the higher metastatic potential of ARMS cells may depend on differential expression of specific matrix metalloproteinases (MMPs) and angiogenesis-related factors, we studied the expression of MMP-2, MT1-MMP, TIMP-2, VEGF and VEGF receptors in four ARMS (RH30, RH4, RH18, RH28), three ERMS (RD, RH36, SMS-CTR) and one undifferentiated sarcoma cell line (A204). Semi-quantitative analysis of MMP-2 revealed high levels of expression in 3 out of 4 ARMS cell lines whereas, among ERMS, only RH36 showed comparable levels of the protease. TIMP-2 and MT1-MMP showed no significant differences among cell lines. in vitro invasiveness was also evaluated. The MMP-2-overexpressing RH30 cells were more invasive than RD cells, which expressed low levels of MMP-2. Exogenous expression of the ARMS specific PAX3-FKHR chimeric protein in RD cells increased MMP-2 activity and invasiveness. Of the three main VEGF isoforms only VEGF165 and VEGF121 were detected in RMS lines: ARMS expressed both isoforms, whereas the ERMS cell line SMS-CTR and the undifferentiated sarcoma cell line A204 showed the VEGF121 isoform only. All RMS cell lines expressed VEGFR-1 at mRNA as well as at protein level. The VEGFR-2, on the contrary, was undetectable with the sole exception of the RH28 cell line. Overall, our data suggest that a high level of MMP-2 protein and VEGF/VEGFR expression may contribute to the metastatic phenotype of ARMS cells and that exogenously induced PAX3-FKHR expression increases MMP-2 secretion and invasive capability of RMS cells.
authors.affiliation: Department of Experimental Biomedical Sciences, University of Padova, I-35121 Padova, Italy. urizio.onisto@unipd.it
authors.forename: Maurizio
authors.initials: M
authors.lastname: Onisto
authors.forename: M Liliana
authors.initials: ML
authors.lastname: Slongo
authors.forename: Laura
authors.initials: L
authors.lastname: Gregnanin
authors.forename: Tamara
authors.initials: T
authors.lastname: Gastaldi
authors.forename: Modesto
authors.initials: M
authors.lastname: Carli
authors.forename: Angelo
authors.initials: A
authors.lastname: Rosolen
mesh_terms: D015153:Blotting, Western; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000235:genetics; D003094:Collagen; D017077:Culture Media, Conditioned / Q000494:pharmacology; D004268:DNA-Binding Proteins / Q000235:genetics; D004338:Drug Combinations; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D000071161:Forkhead Box Protein O1; D051858:Forkhead Transcription Factors; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D007797:Laminin; D020778:Matrix Metalloproteinase 2 / Q000235:genetics / Q000378:metabolism; D053504:Matrix Metalloproteinases, Membrane-Associated; D008666:Metalloendopeptidases / Q000235:genetics / Q000378:metabolism; D045726:Metalloproteases / Q000235:genetics* / Q000378:metabolism; D009361:Neoplasm Invasiveness; D000072041:PAX3 Transcription Factor; D051761:Paired Box Transcription Factors; D010957:Plasmids / Q000235:genetics; D011509:Proteoglycans; D011993:Recombinant Fusion Proteins / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D018232:Rhabdomyosarcoma, Alveolar / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D018233:Rhabdomyosarcoma, Embryonal / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D019716:Tissue Inhibitor of Metalloproteinase-2 / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics; D014162:Transfection; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics* / Q000378:metabolism; D040281:Vascular Endothelial Growth Factor Receptor-1 / Q000235:genetics / Q000378:metabolism; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000235:genetics / Q000378:metabolism
chemical_list: D017077:Culture Media, Conditioned; D004268:DNA-Binding Proteins; D004338:Drug Combinations; C085126:FOXO1 protein, human; D000071161:Forkhead Box Protein O1; D051858:Forkhead Transcription Factors; D007797:Laminin; D000072041:PAX3 Transcription Factor; C489409:PAX3 protein, human; D051761:Paired Box Transcription Factors; D011509:Proteoglycans; D011993:Recombinant Fusion Proteins; D014157:Transcription Factors; D042461:Vascular Endothelial Growth Factor A; C059485:matrigel; D019716:Tissue Inhibitor of Metalloproteinase-2; D003094:Collagen; D040281:Vascular Endothelial Growth Factor Receptor-1; D040301:Vascular Endothelial Growth Factor Receptor-2; D045726:Metalloproteases; D053504:Matrix Metalloproteinases, Membrane-Associated; D008666:Metalloendopeptidases; D020778:Matrix Metalloproteinase 2