Proteomic analysis identified N-cadherin, clusterin, and HSP27 as mediators of SPARC (secreted protein, acidic and rich in cysteines) activity in melanoma cells
Secreted protein, acidic and rich in cysteines (SPARC) is a secreted protein associated with increased aggressiveness of different human cancer types. In order to identify downstream mediators of SPARC activity, we performed a 2-DE proteomic analysis of human melanoma cells following antisense-media...
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paper:paper_16159853_v7_n22_p4123_Sosa2023-06-08T16:25:25Z Proteomic analysis identified N-cadherin, clusterin, and HSP27 as mediators of SPARC (secreted protein, acidic and rich in cysteines) activity in melanoma cells Sosa, María Girotti, María Romina Salvatierra Colussi, Edgardo Enrique Prada, Federico Podhajcer, Osvaldo Luis Melanoma Osteonectin Secreted protein SPARC Two-dimensional gel electrophoresis clusterin complementary RNA cysteine heat shock protein 27 nerve cell adhesion molecule osteonectin protein sparc unclassified drug article cell growth controlled study down regulation human human cell immunoblotting melanoma melanoma cell priority journal protein analysis protein expression protein function proteomics Cadherins Cell Line, Tumor Clusterin Heat-Shock Proteins Humans Melanoma Neoplasm Proteins Osteonectin Proteomics Tumor Cells, Cultured Secreted protein, acidic and rich in cysteines (SPARC) is a secreted protein associated with increased aggressiveness of different human cancer types. In order to identify downstream mediators of SPARC activity, we performed a 2-DE proteomic analysis of human melanoma cells following antisense-mediated downregulation of SPARC expression. We found 23/504 differential spots, 15 of which were identified by peptide fingerprinting analysis. Three of the differential proteins (N-cadherin (N-CAD), clusterin (CLU), and HSP27) were validated by immunoblotting, confirming decreased levels of N-CAD and CLU and increased amounts of HSP27 in conditioned media of cells with diminished SPARC expression. Furthermore, transient knock down of SPARC expression in melanoma cells following adenoviral-mediated transfer of antisense RNA confirmed these changes. We next developed two different RNAs against SPARC that were able to inhibit in vivo melanoma cell growth. Immunoblotting of the secreted fraction of RNAi-transfected melanoma cells confirmed that downregulation of SPARC expression promoted decreased levels of N-CAD and CLU and increased secretion of HSP27. Transient re-expression of SPARC in SPARC-downregulated cells reverted extracellular N-CAD, CLU, and HSP27 to levels similar to those in the control. These results constitute the first evidence that SPARC, N-CAD, CLU, and HSP27 converge in a unique molecular network in melanoma cells. © 2007 WILEY-VCH Verlag GmbH & Co. KGaA. Fil:Sosa, M.S. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Girotti, M.R. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Salvatierra, E. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Prada, F. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Podhajcer, O.L. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 2007 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_16159853_v7_n22_p4123_Sosa http://hdl.handle.net/20.500.12110/paper_16159853_v7_n22_p4123_Sosa |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
Melanoma Osteonectin Secreted protein SPARC Two-dimensional gel electrophoresis clusterin complementary RNA cysteine heat shock protein 27 nerve cell adhesion molecule osteonectin protein sparc unclassified drug article cell growth controlled study down regulation human human cell immunoblotting melanoma melanoma cell priority journal protein analysis protein expression protein function proteomics Cadherins Cell Line, Tumor Clusterin Heat-Shock Proteins Humans Melanoma Neoplasm Proteins Osteonectin Proteomics Tumor Cells, Cultured |
| spellingShingle |
Melanoma Osteonectin Secreted protein SPARC Two-dimensional gel electrophoresis clusterin complementary RNA cysteine heat shock protein 27 nerve cell adhesion molecule osteonectin protein sparc unclassified drug article cell growth controlled study down regulation human human cell immunoblotting melanoma melanoma cell priority journal protein analysis protein expression protein function proteomics Cadherins Cell Line, Tumor Clusterin Heat-Shock Proteins Humans Melanoma Neoplasm Proteins Osteonectin Proteomics Tumor Cells, Cultured Sosa, María Girotti, María Romina Salvatierra Colussi, Edgardo Enrique Prada, Federico Podhajcer, Osvaldo Luis Proteomic analysis identified N-cadherin, clusterin, and HSP27 as mediators of SPARC (secreted protein, acidic and rich in cysteines) activity in melanoma cells |
| topic_facet |
Melanoma Osteonectin Secreted protein SPARC Two-dimensional gel electrophoresis clusterin complementary RNA cysteine heat shock protein 27 nerve cell adhesion molecule osteonectin protein sparc unclassified drug article cell growth controlled study down regulation human human cell immunoblotting melanoma melanoma cell priority journal protein analysis protein expression protein function proteomics Cadherins Cell Line, Tumor Clusterin Heat-Shock Proteins Humans Melanoma Neoplasm Proteins Osteonectin Proteomics Tumor Cells, Cultured |
| description |
Secreted protein, acidic and rich in cysteines (SPARC) is a secreted protein associated with increased aggressiveness of different human cancer types. In order to identify downstream mediators of SPARC activity, we performed a 2-DE proteomic analysis of human melanoma cells following antisense-mediated downregulation of SPARC expression. We found 23/504 differential spots, 15 of which were identified by peptide fingerprinting analysis. Three of the differential proteins (N-cadherin (N-CAD), clusterin (CLU), and HSP27) were validated by immunoblotting, confirming decreased levels of N-CAD and CLU and increased amounts of HSP27 in conditioned media of cells with diminished SPARC expression. Furthermore, transient knock down of SPARC expression in melanoma cells following adenoviral-mediated transfer of antisense RNA confirmed these changes. We next developed two different RNAs against SPARC that were able to inhibit in vivo melanoma cell growth. Immunoblotting of the secreted fraction of RNAi-transfected melanoma cells confirmed that downregulation of SPARC expression promoted decreased levels of N-CAD and CLU and increased secretion of HSP27. Transient re-expression of SPARC in SPARC-downregulated cells reverted extracellular N-CAD, CLU, and HSP27 to levels similar to those in the control. These results constitute the first evidence that SPARC, N-CAD, CLU, and HSP27 converge in a unique molecular network in melanoma cells. © 2007 WILEY-VCH Verlag GmbH & Co. KGaA. |
| author |
Sosa, María Girotti, María Romina Salvatierra Colussi, Edgardo Enrique Prada, Federico Podhajcer, Osvaldo Luis |
| author_facet |
Sosa, María Girotti, María Romina Salvatierra Colussi, Edgardo Enrique Prada, Federico Podhajcer, Osvaldo Luis |
| author_sort |
Sosa, María |
| title |
Proteomic analysis identified N-cadherin, clusterin, and HSP27 as mediators of SPARC (secreted protein, acidic and rich in cysteines) activity in melanoma cells |
| title_short |
Proteomic analysis identified N-cadherin, clusterin, and HSP27 as mediators of SPARC (secreted protein, acidic and rich in cysteines) activity in melanoma cells |
| title_full |
Proteomic analysis identified N-cadherin, clusterin, and HSP27 as mediators of SPARC (secreted protein, acidic and rich in cysteines) activity in melanoma cells |
| title_fullStr |
Proteomic analysis identified N-cadherin, clusterin, and HSP27 as mediators of SPARC (secreted protein, acidic and rich in cysteines) activity in melanoma cells |
| title_full_unstemmed |
Proteomic analysis identified N-cadherin, clusterin, and HSP27 as mediators of SPARC (secreted protein, acidic and rich in cysteines) activity in melanoma cells |
| title_sort |
proteomic analysis identified n-cadherin, clusterin, and hsp27 as mediators of sparc (secreted protein, acidic and rich in cysteines) activity in melanoma cells |
| publishDate |
2007 |
| url |
https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_16159853_v7_n22_p4123_Sosa http://hdl.handle.net/20.500.12110/paper_16159853_v7_n22_p4123_Sosa |
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