Rong Lulu, Wang Jie, Wang Qian, Zhu Yanli, Ren Wenhao
Department of Pathology, Beijing Chao-Yang Hospital, Capital Medical University, Beijing, China.
Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education), Department of Pathology, Peking University Cancer Hospital and Institute, Beijing, China.
Cytojournal. 2025 Feb 14;22:18. doi: 10.25259/Cytojournal_162_2024. eCollection 2025.
The presence of central lymph node metastasis (CLNM) represents a critical determinant in ascertaining the necessity for surgical intervention in patients with papillary thyroid microcarcinoma (PTMC). However, the predominant current methodologies for confirming the central lymph node status in clinical practice are hampered by the low predictive accuracy of preoperative ultrasound examination and the high risk of preoperative fine needle aspiration (FNA). Consequently, the objective of this study is to investigate and identify specific immunocytochemical biomarkers for predicting CLNM in PTMC patients based on preoperative thyroid FNA samples.
In this study, the messenger ribonucleic acid sequencing data of pathological tumor stage 1 (pT1) papillary thyroid carcinoma (PTC) accompanied by pathological node stage information were initially retrieved from The Cancer Genome Atlas database. The differential expression genes (DEGs) between the pT1N1-PTC group and the pT1N0-PTC group were ascertained through bioinformatics methodology. Subsequently, these DEGs were imported into Cytoscape software to identify hub genes. Ultimately, immunohistochemical and immunocytochemical staining were employed to validate whether the biomarkers corresponding to the main hub genes demonstrated statistical significance in predicting CLNM within propensity score-matched PTMC samples.
In this study, a total of 292 DEGs and 10 hub genes were successfully identified. Subsequently, immunohistochemical and immunocytochemical staining were conducted on 208 PTMC cases selected through propensity score matching. Among these 208 cases, the biomarkers (Cytokeratin 5/6 [CK5/6], Chromogranin A [CgA], and Pair box gene 2 [Pax-2]) corresponding to the main hub genes (Cytokeratin 5 [KRT5], Cytokeratin 6 [KRT6A], Chromogranin A [CHGA], and PAX2) were subjected to immunohistochemical staining in postoperative thyroidectomy specimens, the immunohistochemical staining results revealed a statistically significant difference in CK5/6 expression between PTMCs with and without CLNM ( = 0.002). Subsequently, CK5/6 immunocytochemical staining performed on preoperative thyroid FNA liquid-based samples further corroborated that CK5/6 expression was more prone to being positive in PTMCs with CLNM ( = 0.010).
CK5/6 is a valuable immunocytochemical biomarker capable of predicting the occurrence of CLNM in PTMC patients prior to surgery.
中央区淋巴结转移（CLNM）的存在是确定甲状腺微小乳头状癌（PTMC）患者是否需要手术干预的关键决定因素。然而，目前临床实践中用于确认中央区淋巴结状态的主要方法存在术前超声检查预测准确性低和术前细针穿刺活检（FNA）风险高的问题。因此，本研究的目的是基于术前甲状腺FNA样本，研究并确定预测PTMC患者CLNM的特异性免疫细胞化学生物标志物。
在本研究中，首先从癌症基因组图谱数据库中检索出伴有病理淋巴结分期信息的病理肿瘤1期（pT1）甲状腺乳头状癌（PTC）的信使核糖核酸测序数据。通过生物信息学方法确定pT1N1-PTC组和pT1N0-PTC组之间的差异表达基因（DEGs）。随后，将这些DEGs导入Cytoscape软件以识别核心基因。最后，采用免疫组织化学和免疫细胞化学染色来验证与主要核心基因对应的生物标志物在倾向评分匹配的PTMC样本中预测CLNM是否具有统计学意义。
在本研究中，共成功鉴定出292个DEGs和10个核心基因。随后，对通过倾向评分匹配选择的208例PTMC病例进行了免疫组织化学和免疫细胞化学染色。在这208例病例中，对与主要核心基因（细胞角蛋白5 [KRT5]、细胞角蛋白6 [KRT6A]、嗜铬粒蛋白A [CHGA]和配对盒基因2 [PAX2]）对应的生物标志物（细胞角蛋白5/6 [CK5/6]、嗜铬粒蛋白A [CgA]和配对盒基因2 [Pax-2]）在术后甲状腺切除标本中进行免疫组织化学染色，免疫组织化学染色结果显示，有CLNM和无CLNM的PTMC之间CK5/6表达存在统计学显著差异（ = 0.002）。随后，对术前甲状腺FNA液基样本进行的CK5/6免疫细胞化学染色进一步证实，CLNM的PTMC中CK5/6表达更易呈阳性（ = 0.010）。
CK5/6是一种有价值的免疫细胞化学生物标志物，能够在手术前预测PTMC患者CLNM的发生。
