Abstract
Smad4 is a tumor-suppressor gene that is lost or mutated in 50% of pancreatic carcinomas. Smad4 is also an intracellular transmitter of transforming growth factor-β (TGF-β) signals. Although its tumor-suppressor function is presumed to reside in its capacity to mediate TGF-β-induced growth inhibition, there seems to be a Smad4-independent TGF-β signaling pathway. Here, we succeeded in establishing Smad4 knockdown (S4KD) pancreatic cancer cell lines using stable RNA interference. Smad4 protein expression and TGF-β-Smad4 signaling were impaired in S4KD cells, and we compared the proteomic changes with TGF-β stimulation using two-dimensional gel electrophoresis (2-DE) and mass spectrometry. We identified five proteins that were up-regulated and seven proteins that were down-regulated; 10 of them were novel targets for TGF-β. These proteins function in processes such as cytoskeletal regulation, cell cycle, and oxidative stress. Introducing siRNA-mediated gene silencing into proteomics revealed a novel TGF-β signal pathway that did not involve Smad4.
Original language | English (US) |
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Pages (from-to) | 289-296 |
Number of pages | 8 |
Journal | Biochemical and Biophysical Research Communications |
Volume | 318 |
Issue number | 1 |
DOIs | |
State | Published - May 21 2004 |
Externally published | Yes |
Keywords
- 2-DE
- Proteomics
- Smad4
- Stable RNAi
- TGF-β
ASJC Scopus subject areas
- Biophysics
- Biochemistry
- Molecular Biology
- Cell Biology