目的:探讨细胞周期蛋白D2(CCND2)在甲状腺乳头状癌(PTC)中的表达及其与临床病理特征的关系。
方法:公共数据库TCGA,用TIMER2.0和UALCAN探讨CCND2在PTC和邻近组织中的表达水平,用ROC曲线分析其对PTC的诊断价值。对PTC中CCND2相关差异表达基因(DEGs)进行GO富集分析,使用TIMER数据库和CIBERSORT数据源分析甲状腺癌中CCND2的肿瘤免疫浸润。RT-qPCR和Westernblot检测CCND2在正常人甲状腺细胞系Nthy-ori-3-1和人PTC细胞系TPC-1和BCPAP中的表达。用免疫组化法检测PTC临床标本及癌旁组织中CCND2的表达,并分析其与患者临床病理特征的相关性。
结果:信息学分析显示,甲状腺癌中CCND2mRNA的表达明显高于癌旁组织(P<0.001),与肿瘤分期密切相关。性别,年龄,病理亚型,淋巴结受累(P<0.05)。ROC曲线分析表明,在截断值4.983时,特异性,PTC的CCND2表达准确率为83.6%,94.9%,78.5%,分别。CCND2表达与B细胞呈正相关,CD4+T细胞,与巨噬细胞(P<0.001)和CD8+T细胞(P<0.01),还与记忆B细胞浸润有关,CD4+T细胞记忆激活,M2巨噬细胞,静息的肥大细胞,肥大细胞活化(P<0.05)。RT-qPCR,Westernblot和免疫组化显示,与肿瘤大小相关的PTC细胞中CCND2的表达明显高于Nthy-ori-3-1细胞(P<0.01),临床PTC组织中CCND2的表达也明显高于癌旁组织(P<0.05)。淋巴结转移和TNM分期(P<0.05)。
结论:CCND2过表达与PTC患者肿瘤进展和免疫细胞浸润密切相关。.
OBJECTIVE: To investigate cyclin D2 (CCND2) expression in papillary thyroid carcinoma (PTC) and its association with the clinicopathological features.
METHODS: The public databases TCGA, TIMER 2.0 and UALCAN were used to explore CCND2 expression level in PTC and adjacent tissues, and its diagnostic value for PTC was analyzed using ROC curves. GO enrichment analysis of CCND2-related differentially expressed genes (DEGs) in PTC was performed, and tumor immune infiltration of CCND2 in thyroid cancer was analyzed using TIMER database and CIBERSORT data source. RT-qPCR and Western blot were used to detect CCND2 expression in normal human thyroid cell line Nthy-ori-3-1 and human PTC cell lines TPC-1 and BCPAP. CCND2 expression was also detected in clinical specimens of PTC and adjacent tissues by immunohistochemistry, and its correlation with clinicopathological features of the patients were analyzed.
RESULTS: Informatic analysis revealed significantly higher CCND2 mRNA expression in thyroid cancer than in the adjacent tissues (P < 0.001) in close correlation with tumor stage, gender, age, pathological subtype, and lymph node involvement (P < 0.05). ROC curve analysis showed that at the cutoff value of 4.983, the diagnostic sensitivity, specificity, and accuracy of CCND2 expression for PTC was 83.6%, 94.9%, and 78.5%, respectively. CCND2 expression was positively correlated with B cells, CD4+ T cells, and macrophages (P < 0.001) and negatively with CD8+ T cells (P < 0.01), and also correlated with memory B-cell infiltration, CD4+ T-cell memory activation, M2 macrophages, resting mast cells, and mast cell activation (P < 0.05). RT-qPCR, Western blot and immunohistochemistry showed significantly higher CCND2 expression in the PTC cells than in Nthy-ori-3-1 cells (P < 0.01) and also in clinical PTC tissues than in the adjacent tissues (P < 0.05) in correlation with tumor size, lymph node metastasis and TNM stage (P < 0.05).
CONCLUSIONS: CCND2 overexpression is closely correlated with tumor progression and immune cell infiltration in PTC patients..