Identification of TGFβ-induced proteins in non-endocrine mouse pituitary cell line TtT/GF by SILAC-assisted quantitative mass spectrometry
Identification of TGFβ-induced proteins in non-endocrine mouse pituitary cell line TtT/GF by SILAC-assisted quantitative mass spectrometry
复制标题
SILAC 定量辅助质谱法鉴定非内分泌小鼠垂体细胞系 TtT/GF 中 TGFβ 诱导的蛋白
DOI：
10.1007/s00441-018-02989-2
复制
发表时间：
2019
影响因子：
3.6
通讯作者：
Kato Yukio
中科院分区：
文献类型：
--
作者：
Tsukada Takehiro;Isowa Yukinobu;Kito Keiji;Yoshida Saishu;Toneri Seina;Horiguchi Kotaro;Fujiwara Ken;Yashiro Takashi;Kato Takako;Kato Yukio
TtT/GF是一种来源于促甲状腺垂体瘤的小鼠细胞系，已被用作滤泡星状细胞的模型。我们先前的芯片数据表明TtT/GF具有内皮细胞、周细胞和干/祖细胞的某些特性，沿着有滤泡星状细胞，提示其具有可塑性。我们还发现转化生长因子β（TGFβ）改变TtT/GF细胞中的细胞运动性，增加周细胞标志物转录，并减弱内皮细胞和干/祖细胞标志物。本研究在蛋白水平上探索TGFβ对TtT/GF细胞的广泛影响，并使用细胞培养中氨基酸的稳定同位素标记（SILAC）辅助定量质谱来表征TGFβ诱导的蛋白及其伙伴关系。比较TGFβ处理的细胞和SB 431542（一种选择性TGFβ受体I抑制剂）处理的细胞的定量蛋白质，发现51种蛋白质上调，112种蛋白质下调。|log2|> 0.6）。基因本体和STRING分析表明，这些与肌动蛋白细胞骨架，细胞粘附，细胞外基质和DNA复制有关。一致地，TGFβ处理的细胞显示出独特的肌动蛋白丝模式，并且与媒介物处理的细胞相比增殖减少; SB 431542阻断TGFβ的作用。许多周细胞标志物（CSPG 4、内斯、ACTA、TAGLN、COL 1A 1、THBS 1、TIMP 3和FLNA）的上调支持我们先前的假设，即TGFβ增强周细胞性质。我们还发现CTSB、EZR和LGALS 3的下调，这在几种垂体腺瘤中被诱导。这些数据提供了有价值的信息，周细胞分化以及垂体腺瘤的病理过程。
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.