Abstract：
Thyroid cancer has emerged as the most rapidly proliferating solid neoplasm. In this study, we included a cohort of patients who underwent sonographic assessment and surgical intervention at the Sir Run Run Shaw Hospital, associated with the School of Medicine at Zhejiang University, spanning from January 2019 to June 2020. Stratification of cases was based on a combination of preoperative ultrasonographic evaluations and postoperative histopathological diagnoses, resulting in three distinct groups: high-risk papillary thyroid carcinoma (PTC) labeled as C1, low-risk PTC designated as C2, and a control group (N) composed of benign thyroid tissue adjacent to the carcinoma. Proteomic and phosphoproteomic analyses were conducted on PTC specimens. The comparative assessment revealed that proteins up-regulated in the C1/N and C2/N groups were predominantly involved in functions such as amino acid binding, binding of phosphorylated compounds, and serine protease activity. Notably, proteins like NADH dehydrogenase, ATP synthase, oxidoreductases, and iron ion channels were significantly elevated in the C1 versus C2 comparative group. Through meticulous analysis of differential expression multiples, statistical significance, and involvement in metabolic pathways, this study identified eight potential biomarkers pertinent to PTC metastasis diagnostics, encompassing phosphorylated myosin 10, phosphorylated proline-directed protein kinase, leucine tRNA synthetase, 2-oxo-isovalerate dehydrogenase, succinic semialdehyde dehydrogenase, ADP/ATPtranslocase, pyruvate carboxylase, and fibrinogen. Therapeutic assays employing metformin, an AMP-activated protein kinase (AMPK) activator, alongside the phosphorylation-specific inhibitor ML-7 targeting Myosin10, demonstrated attenuated cellular proliferation, migration, and invasion capabilities in thyroid cancer cells, accompanied by a reduction in amino acid pools. Cellular colocalization and interaction studies elucidated that AMPK activation imposes an inhibitory influence on Myosin10 levels. The findings of this research corroborate the utility of proteomic and phosphoproteomic platforms in the identification of metastatic markers for PTC and suggest that modulation of AMPK activity, coupled with the inhibition of Myosin10 phosphorylation, may forge novel therapeutic avenues in the management of thyroid carcinoma. SIGNIFICANCE: The significance of our research lies in its potential to transform the current understanding and management of thyroid papillary carcinoma (PTC), particularly in its metastatic form. By integrating both proteomic and phosphoproteomic analyses, our study not only sheds light on the molecular alterations associated with PTC but also identifies eight novel biomarkers that could serve as indicators of metastatic potential.
摘要：
甲状腺癌已成为增殖最快的实体瘤。在这项研究中,我们纳入了一组在SirRunRunShaw医院接受超声检查和手术干预的患者，与浙江大学医学院有关，从2019年1月到2020年6月。病例的分层是基于术前超声评估和术后组织病理学诊断的组合，结果分为三个不同的组：标记为C1的高风险甲状腺乳头状癌（PTC），标记为C2的低风险PTC和由邻近癌的良性甲状腺组织组成的对照组（N）。对PTC标本进行蛋白质组学和磷酸蛋白质组学分析。比较评估显示，在C1/N和C2/N组中上调的蛋白质主要参与氨基酸结合等功能。磷酸化化合物的结合，和丝氨酸蛋白酶活性。值得注意的是,像NADH脱氢酶这样的蛋白质，ATP合成酶,氧化还原酶，和铁离子通道在C1和C2比较组中显著升高。通过对差异表达倍数的细致分析,统计意义，参与代谢途径，这项研究确定了八个潜在的生物标志物相关的PTC转移诊断，包括磷酸化肌球蛋白10，磷酸化脯氨酸定向蛋白激酶，亮氨酸tRNA合成酶，2-氧代-异戊酸脱氢酶，琥珀酸半醛脱氢酶，ADP/ATP转位酶，丙酮酸羧化酶，和纤维蛋白原。使用二甲双胍的治疗试验，AMP激活的蛋白激酶（AMPK）激活剂，与针对Myosin10的磷酸化特异性抑制剂ML-7一起，证明细胞增殖减弱，迁移,和甲状腺癌细胞的侵袭能力，伴随着氨基酸池的减少。细胞共定位和相互作用研究阐明了AMPK激活对Myosin10水平具有抑制作用。这项研究的结果证实了蛋白质组学和磷酸蛋白质组学平台在鉴定PTC转移标志物中的实用性，并表明AMPK活性的调节，再加上抑制Myosin10磷酸化，可能为甲状腺癌的治疗开辟新的治疗途径。意义：我们的研究的意义在于它有可能改变目前对甲状腺乳头状癌（PTC）的理解和管理，特别是其转移形式。通过整合蛋白质组学和磷酸化蛋白质组学分析，我们的研究不仅揭示了与PTC相关的分子改变,而且还确定了8种可作为转移潜能指标的新型生物标志物.
