Proteomic analysis of differentially expressed proteins in normal human thyroid cells transfected with PPFP
Proteomic analysis of differentially expressed proteins in normal human thyroid cells transfected with PPFP
复制标题
转染 PPFP 的正常人甲状腺细胞差异表达蛋白的蛋白质组学分析。
DOI：
10.1530/erc-12-0156
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发表时间：
2012-10-01
影响因子：
3.9
通讯作者：
Li, Cui
中科院分区：
文献类型：
--
作者：
Li, Xinying;Wang, Zhiming;Li, Cui
编码甲状腺特异性转录因子 PAX8 和过氧化物酶体增殖物激活受体 γ (PPAR γ (PPARG)) 的融合基因（称为 PPFP 基因）具有致癌性，与滤泡性甲状腺癌 (FTC) 的发展有关。通过MTT法、集落形成法、软琼脂集落形成法、划痕伤口愈合试验以及流式细胞术研究PPFP转染对Nthy-ori 3-1细胞生物学特性的影响。此外，在 2-DE 图谱上分析差异表达的蛋白质，并通过 MALDI-TOF-MS 进行鉴定。通过蛋白质印迹分析确定五种已鉴定蛋白质（抑制素、半乳糖凝集素-1、细胞角蛋白 8 (CK8)、CK19 和 HSP27）的验证。 PPFP不仅显着增加Nthy-ori 3-1细胞的活力、增殖和迁移率，而且显着抑制细胞凋亡。鉴定出28个差异表达蛋白，其中Nthy-ori 3-1(PPFP)（转染PPFP的Nthy-ori 3-1细胞）中19个蛋白上调，9个蛋白下调。 Western blot结果与蛋白质组分析结果一致，显示Nthy-ori 3-1（PPFP）中prohibitin下调，而galectin-1、CK8、CK19和HSP27上调。我们的结果表明 PPFP 在甲状腺恶性转化中发挥重要作用。对PPFP转染细胞中差异表达蛋白进行蛋白质组学分析，为进一步研究FTC中PPFP的致癌机制提供了重要信息。内分泌相关癌症（2012）19 681-694
The fusion gene encoding the thyroid-specific transcription factor PAX8 and peroxisome proliferator-activated receptor gamma (PPAR gamma (PPARG)) (designated as the PPFP gene) is oncogenic and implicated in the development of follicular thyroid carcinoma (FTC). The effects of PPFP transfection on the biological characteristics of Nthy-ori 3-1 cells were studied by MTT assay, colony formation, soft-agar colony formation, and scratch wound-healing assays as well as by flow cytometry. Furthermore, the differentially expressed proteins were analyzed on 2-DE maps and identified by MALDI-TOF-MS. Validation of five identified proteins (prohibitin, galectin-1, cytokeratin 8 (CK8), CK19, and HSP27) was determined by western blot analysis. PPFP not only significantly increased the viability, proliferation, and mobility of the Nthy-ori 3-1 cells but also markedly inhibited cellular apoptosis. Twenty-eight differentially expressed proteins were identified, among which 19 proteins were upregulated and nine proteins were downregulated in Nthy-ori 3-1(PPFP) (Nthy-ori 3-1 cells transfected with PPFP). The western blot results, which were consistent with the proteome analysis results, showed that prohibitin was downregulated, whereas galectin-1, CK8, CK19, and HSP27 were upregulated in Nthy-ori 3-1(PPFP). Our results suggest that PPFP plays an important role in malignant thyroid transformation. Proteomic analysis of the differentially expressed proteins in PPFP-transfected cells provides important information for further study of the carcinogenic mechanism of PPFP in FTCs. Endocrine-Related Cancer (2012) 19 681-694