Wei Wei, Wu Yuezhang, Chen Dong-Dong, Song Yuntao, Xu Guohui, Shi Qi, Dong Xiao-Ping
Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education), Head and Neck Surgery Department, Peking University Cancer Hospital & Institute, Beijing, 100142, China.
State Key Laboratory for Infectious Disease Prevention and Control, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases (Zhejiang University), National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Chang-Bai Rd 155, Beijing, 102206, China.
Proteome Sci. 2023 Apr 26;21(1):6. doi: 10.1186/s12953-023-00207-8.
Papillary thyroid carcinoma (PTC) is the most common endocrine malignancy cancer among the malignancies of thyroid. Despite of wide usages of proteomics in PTC, the profile of acetylated proteins in PTC remains unsettled, which is helpful for understanding the carcinogenesis mechanism and identifying useful biomarkers for PTC.
The surgically removed specimens of cancer tissues (Ca-T) and adjacent normal tissues (Ca-N) from 10 female patients pathological diagnosed as PTC (TNM stage III) were enrolled in the study. After preparing the pooled extracts of the whole proteins and the acetylated proteins from 10 cases, TMT labeling and LC/MS/MS methods were applied to the assays of global proteomics and acetylated proteomics separately. Bioinformatics analysis, including KEGG, gene ontology (GO) and hierarchical clustering were performed. Some differentially expressed proteins (DEPs) and differentially expressed acetylated proteins (DEAPs) were validated by individual Western blots.
Controlled with the normal tissues adjacent to the lesions, 147 out of 1923 identified proteins in tumor tissues were considered as DEPs in global proteomics, including 78 up-regulated and 69 down-regulated ones, while 57 out of 311 identified acetylated proteins in tumor tissues were DEAPs in acetylated proteomics, including 32 up-regulated and 25 down-regulated, respectively. The top 3 up- and down-regulated DEPs were fibronectin 1, KRT1B protein and chitinase-3-like protein 1, as well as keratin, type I cytoskeletal 16, A-gamma globin Osilo variant and Huntingtin interacting protein-1. The top 3 up- and down-regulated DEAPs were ribosomal protein L18a-like protein, alpha-1-acid glycoprotein 2 and eukaryotic peptide chain release factor GTP-binding subunit ERF3A, as well as trefoil factor 3, thyroglobulin and histone H2B. Functional GO annotation and KEGG pathway analysis based on the DEPs and DEAPs showed completely different changing pictures. Contrary to the top 10 up- and -down regulated DEPs, most of which were addressed in PTC and other types of carcinomas, changes of the majority DEAPs were not mentioned in the literatures.
Taken the profiling of the global and acetylated proteomics together will provide more broad view of protein alterations on the carcinogenesis and new direction for selecting biomarker for diagnosis of PTC.
甲状腺乳头状癌(PTC)是甲状腺恶性肿瘤中最常见的内分泌恶性肿瘤。尽管蛋白质组学在PTC研究中应用广泛,但PTC中乙酰化蛋白质的概况仍未明确,这有助于理解其致癌机制并识别PTC的有用生物标志物。
本研究纳入了10例经病理诊断为PTC(TNM III期)的女性患者手术切除的癌组织(Ca-T)和癌旁正常组织(Ca-N)标本。制备10例样本的全蛋白和乙酰化蛋白混合提取物后,分别采用TMT标记和LC/MS/MS方法进行整体蛋白质组学和乙酰化蛋白质组学分析。进行了包括KEGG、基因本体论(GO)和层次聚类在内的生物信息学分析。部分差异表达蛋白(DEPs)和差异表达乙酰化蛋白(DEAPs)通过个体蛋白质免疫印迹法进行验证。
与病变旁正常组织相比,肿瘤组织中1923个鉴定出的蛋白质中有147个被视为整体蛋白质组学中的DEPs,其中78个上调,69个下调;而肿瘤组织中311个鉴定出的乙酰化蛋白质中有57个为乙酰化蛋白质组学中的DEAPs,分别为32个上调和25个下调。上调和下调的前3个DEPs分别是纤连蛋白1、角蛋白1B和几丁质酶3样蛋白1,以及角蛋白I型细胞骨架16、A-γ球蛋白Osilo变体和亨廷顿相互作用蛋白1。上调和下调的前3个DEAPs分别是核糖体蛋白L18a样蛋白、α-1-酸性糖蛋白2和真核肽链释放因子GTP结合亚基ERF3A,以及三叶因子3、甲状腺球蛋白和组蛋白H2B。基于DEPs和DEAPs的功能GO注释和KEGG通路分析显示出完全不同的变化图景。与上调和下调的前十大DEPs相反,它们大多在PTC和其他类型的癌症中被提及,而大多数DEAPs的变化在文献中未被提及。
综合整体蛋白质组学和乙酰化蛋白质组学分析将为致癌过程中的蛋白质变化提供更全面的视角,并为PTC诊断生物标志物的选择提供新方向。