Identification of TGFβ-induced proteins in non-endocrine mouse pituitary cell line TtT/GF by SILAC-assisted quantitative mass spectrometry
| Autor: | Yukio Kato, Takehiro Tsukada, Yukinobu Isowa, Seina Toneri, Takako Kato, Takashi Yashiro, Kotaro Horiguchi, Keiji Kito, Saishu Yoshida, Ken Fujiwara |
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| Rok vydání: | 2019 |
| Předmět: |
Proteomics
0301 basic medicine Histology Cell Plasticity DNA-Directed DNA Polymerase Mass Spectrometry Pathology and Forensic Medicine Extracellular matrix Mice 03 medical and health sciences 0302 clinical medicine Multienzyme Complexes Pituitary Gland Anterior Transforming Growth Factor beta Cell Line Tumor Stable isotope labeling by amino acids in cell culture medicine Animals Progenitor cell Cell adhesion Extracellular Matrix Proteins biology Chemistry Cell Biology Transforming growth factor beta Actin cytoskeleton Cell biology Endothelial stem cell Cytoskeletal Proteins 030104 developmental biology medicine.anatomical_structure Isotope Labeling biology.protein Pericyte Pericytes Cell Adhesion Molecules 030217 neurology & neurosurgery |
| Zdroj: | Cell and Tissue Research. 376:281-293 |
| ISSN: | 1432-0878 0302-766X |
| Popis: | TtT/GF is a mouse cell line derived from a thyrotropic pituitary tumor and has been used as a model of folliculostellate cells. Our previous microarray data indicate that TtT/GF possesses some properties of endothelial cells, pericytes and stem/progenitor cells, along with folliculostellate cells, suggesting its plasticity. We also found that transforming growth factor beta (TGFβ) alters cell motility, increases pericyte marker transcripts and attenuates endothelial cell and stem/progenitor cell markers in TtT/GF cells. The present study explores the wide-range effect of TGFβ on TtT/GF cells at the protein level and characterizes TGFβ-induced proteins and their partnerships using stable isotope labeling of amino acids in cell culture (SILAC)-assisted quantitative mass spectrometry. Comparison between quantified proteins from TGFβ-treated cells and those from SB431542 (a selective TGFβ receptor I inhibitor)-treated cells revealed 51 upregulated and 112 downregulated proteins (|log2| > 0.6). Gene ontology and STRING analyses revealed that these are related to the actin cytoskeleton, cell adhesion, extracellular matrix and DNA replication. Consistently, TGFβ-treated cells showed a distinct actin filament pattern and reduced proliferation compared to vehicle-treated cells; SB431542 blocked the effect of TGFβ. Upregulation of many pericyte markers (CSPG4, NES, ACTA, TAGLN, COL1A1, THBS1, TIMP3 and FLNA) supports our previous hypothesis that TGFβ reinforces pericyte properties. We also found downregulation of CTSB, EZR and LGALS3, which are induced in several pituitary adenomas. These data provide valuable information about pericyte differentiation as well as the pathological processes in pituitary adenomas. |
| Databáze: | OpenAIRE |
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