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Proteomic analysis identified N-cadherin, clusterin, and HSP27 as mediators of SPARC (secreted protein, acidic and rich in cysteines) activity in melanoma cells

Dr.LinZhen 添加于 2010-11-28 22:22 | 995 次阅读 | 0 个评论

作者
Sosa MS, Girotti MR, Salvatierra E, Prada F, de Olmo JAL, Gallango SJ, Albar JP, Podhajcer OL, Llera AS
摘要
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.
详细资料
- 关键词: Cadherins/analysis/*metabolism; Cell Line, Tumor; Clusterin/analysis/*metabolism; HSP27 Heat-Shock Proteins; Heat-Shock Proteins/analysis/*metabolism; Humans; Melanoma/*metabolism; Neoplasm Proteins/analysis/*metabolism; Osteonectin/biosynthesis/*metabolism; Proteomics/methods; Tumor Cells, Cultured
- 文献种类:期刊
- 期刊名称: Proteomics
- 期刊缩写: Proteomics
- 期卷页: 2007年第7卷第22期 4123-4134页
- 地址: Laboratory of Molecular and Cellular Therapy, Fundacion Instituto Leloir-CONICET and Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Buenos Aires, Argentina
- ISBN: 1615-9853
- 备注:PMID:17994631
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