Joseph and Mildred Sonshine Family Centre for Head and Neck Diseases, Mount Sinai Hospital, Toronto, Ontario, Canada.
J Proteome Res. 2010 Nov 5;9(11):5757-69. doi: 10.1021/pr100529t. Epub 2010 Oct 15.
In search of thyroid cancer biomarkers, proteins secreted by thyroid cancer cell lines, papillary-derived TPC-1 and anaplastic-derived CAL62, were analyzed using liquid chromatography-tandem mass spectrometry. Of 46 high-confidence identifications, 6 proteins were considered for verification in thyroid cancer patients' tissue and blood. The localization of two proteins, nucleolin and prothymosin-α (PTMA), was confirmed in TPC-1 and CAL62 cells by confocal microscopy and immunohistochemically in xenografts of TPC-1 cells in NOD/SCID/γ mice and human thyroid cancers (48 tissues). Increased nuclear and cytoplasmic expression of PTMA was observed in anaplastic compared to papillary and poorly differentiated carcinomas. Nuclear expression of nucleolin was observed in all subtypes of thyroid carcinomas, along with faint cytoplasmic expression in anaplastic cancers. Importantly, PTMA, nucleolin, clusterin, cysteine-rich angiogenic inducer 61, enolase 1, and biotinidase were detected in thyroid cancer patients' sera, warranting future analysis to confirm their potential as blood-based thyroid cancer markers. In conclusion, we demonstrated the potential of secretome analysis of thyroid cancer cell lines to identify novel proteins that can be independently verified in cell lines, xenografts, tumor tissues, and blood samples of thyroid cancer patients. These observations support their potential utility as minimally invasive biomarkers for thyroid carcinomas and their application in management of these diseases upon future validation.
为了寻找甲状腺癌生物标志物，我们使用液相色谱-串联质谱法分析了甲状腺癌细胞系（乳头状来源的 TPC-1 和间变性来源的 CAL62）分泌的蛋白质。在 46 种高可信度鉴定中，有 6 种蛋白质被认为可在甲状腺癌患者的组织和血液中进行验证。通过共聚焦显微镜和免疫组织化学，在 NOD/SCID/γ 小鼠异种移植 TPC-1 细胞和人甲状腺癌（48 种组织）中，证实了核仁蛋白和前胸腺素-α（PTMA）这两种蛋白质在 TPC-1 和 CAL62 细胞中的定位。与乳头状和低分化癌相比，间变性癌中观察到 PTMA 的核内和细胞质表达增加。核仁蛋白在所有甲状腺癌亚型中均有表达，间变性癌中则有微弱的细胞质表达。重要的是，PTMA、核仁蛋白、簇蛋白、富含半胱氨酸的血管生成诱导因子 61、烯醇酶 1 和生物素酶在甲状腺癌患者的血清中被检测到，这需要进一步分析来确认它们作为基于血液的甲状腺癌标志物的潜力。总之，我们证明了甲状腺癌细胞系分泌组分析鉴定新蛋白的潜力，这些蛋白可以在细胞系、异种移植瘤、肿瘤组织和甲状腺癌患者的血液样本中独立验证。这些观察结果支持它们作为甲状腺癌微创生物标志物的潜在用途，并支持在未来验证后将其应用于这些疾病的管理。
