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Published on: September 25, 2011
Novel snail1 target proteins in human colon cancer identified by proteomic analysis
María Jesús Larriba1, Juan Casado-Vela, Natalia Pendás-Franco
1Instituto de Investigaciones Biomédicas Alberto Sols, Consejo Superior de Investigaciones Científicas-Universidad Autónoma de Madrid, Madrid, Spain. mjlarriba@iib.uam.es
Background:
The transcription factor Snail1 induces epithelial-to-mesenchymal transition (EMT), a process responsible for the acquisition of invasiveness during tumorigenesis. Several transcriptomic studies have reported Snail1-regulated genes in different cell types, many of them involved in cell adhesion. However, only a few studies have used proteomics as a tool for the characterization of proteins mediating EMT.
Methodology/Principal Findings:
We identified by proteomic analysis using 2D-DIGE electrophoresis combined with MALDI-TOF-TOF and ESI-linear ion trap mass spectrometry a number of proteins with variable functions whose expression is modulated by Snail1 in SW480-ADH human colon cancer cells. Validation was performed by Western blot and immunofluorescence analyses. Snail1 repressed several members of the 14-3-3 family of phosphoserine/phosphothreonine binding proteins and also the expression of the Proliferation-associated protein 2G4 (PA2G4) that was mainly localized at the nuclear Cajal bodies. In contrast, the expression of two proteins involved in RNA processing, the Cleavage and polyadenylation specificity factor subunit 6 (CPSF6) and the Splicing factor proline/glutamine-rich (SFPQ), was higher in Snail1-expressing cells than in controls. The regulation of 14-3-3epsilon, 14-3-3tau, 14-3-3zeta and PA2G4 by Snail1 was reproduced in HT29 colon cancer cells. In addition, we found an inverse correlation between 14-3-3sigma and Snail1 expression in human colorectal tumors.
Conclusions/Significance:
We have identified a set of novel Snail1 target proteins in colon cancer that expand the cellular processes affected by Snail1 and thus its relevance for cell function and phenotype.
Insights
This study identifies novel Snail1 target proteins in colon cancer cells, revealing new insights into how Snail1 influences cell function and invasiveness during tumorigenesis.
Area of Science:
- Oncology
- Molecular Biology
- Proteomics
Background:
- The transcription factor Snail1 drives epithelial-to-mesenchymal transition (EMT), a key process in cancer cell invasion.
- While transcriptomic studies exist, few proteomic analyses have explored proteins mediating EMT.
- Understanding Snail1's protein targets is crucial for elucidating its role in tumorigenesis.
Purpose of the Study:
- To identify novel proteins regulated by Snail1 in human colon cancer cells using a proteomic approach.
- To characterize the functional impact of Snail1-modulated proteins on cellular processes.
- To validate Snail1's regulation of specific protein targets in different colon cancer cell lines and human tumors.
Main Methods:
- Proteomic analysis using 2D-DIGE electrophoresis coupled with MALDI-TOF-TOF and ESI-linear ion trap mass spectrometry.
- Validation of protein expression changes via Western blot and immunofluorescence.
- Correlation analysis of protein expression with Snail1 levels in human colorectal tumors.
Main Results:
- Snail1 modulated the expression of various proteins in SW480-ADH colon cancer cells.
- Snail1 repressed 14-3-3 family proteins and Proliferation-associated protein 2G4 (PA2G4).
- Snail1 upregulated RNA processing factors CPSF6 and SFPQ; regulation confirmed in HT29 cells and inversely correlated with 14-3-3sigma in tumors.
Conclusions:
- Identified novel Snail1 target proteins in colon cancer, expanding understanding of Snail1's cellular targets.
- These findings highlight new cellular processes affected by Snail1, impacting cell function and phenotype.
- The study provides a proteomic basis for Snail1's role in colon cancer progression and invasiveness.
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