Key Laboratory of Cancer Proteomics of Chinese Ministry of Health, Xiangya Hospital, Central South University, Changsha, People's Republic of China.
Endocr Relat Cancer. 2012 Sep 21;19(5):681-94. doi: 10.1530/ERC-12-0156. Print 2012 Oct.
The fusion gene encoding the thyroid-specific transcription factor PAX8 and peroxisome proliferator-activated receptor γ (PPARγ (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.
PAX8 和过氧化物酶体增殖物激活受体 γ（PPARγ（PPARG））的甲状腺特异性转录因子融合基因（命名为 PPFP 基因）是致癌的，并与滤泡甲状腺癌（FTC）的发展有关。通过 MTT 测定、集落形成、软琼脂集落形成、划痕愈合试验和流式细胞术研究了 PPFP 转染对 Nthy-ori 3-1 细胞生物学特性的影响。此外，通过 2-DE 图谱分析和 MALDI-TOF-MS 鉴定差异表达的蛋白质。通过 Western blot 分析验证了五个鉴定蛋白（抑制素、半乳糖凝集素-1、细胞角蛋白 8（CK8）、CK19 和 HSP27）。PPFP 不仅显著增加了 Nthy-ori 3-1 细胞的活力、增殖和迁移能力，而且显著抑制了细胞凋亡。鉴定出 28 种差异表达蛋白，其中 Nthy-ori 3-1（PPFP）（转染 PPFP 的 Nthy-ori 3-1 细胞）中 19 种蛋白上调，9 种蛋白下调。Western blot 结果与蛋白质组分析结果一致，表明抑制素下调，而半乳糖凝集素-1、CK8、CK19 和 HSP27 上调。我们的结果表明，PPFP 在恶性甲状腺转化中起重要作用。PPFP 转染细胞差异表达蛋白的蛋白质组学分析为进一步研究 PPFP 在 FTC 中的致癌机制提供了重要信息。
