目的： 通过对比分析乳头状甲状腺癌（papillary thyroid cancer,PTC）和癌旁组织中NF-κB信号通路相关基因的差异表达,以及52个实体瘤相关基因的突变情况,鉴定与PTC临床诊断治疗相关的肿瘤特异性分子靶点。方法： 分别提取20例PTC石蜡样本的癌与癌旁组织的RNA及DNA,运用定量反转录聚合酶链式反应（quantitative reverse transcription PCR,qRT-PCR）分析NF-κB信号通路目的基因CD44、BCL2、CCND2、c-FLIP、IκBα、A20及ABINs的RNA水平表达,并用免疫组化方法进行蛋白水平表达验证。同时利用靶向二代测序（next generation sequencing,NGS）对其中5例随访复发病例的肿瘤相关基因的突变情况进行全面分析筛选。结果： PTC癌组织中CD44和CCND2基因在RNA和蛋白水平的表达均显著高于癌旁组织。在有淋巴结转移和无淋巴结转移的PTC间,各NF-κB目的基因的表达量差异均无统计学意义。在基因水平,ALK、BRAF、FGFR3/4、KIT、MYC及MAPK信号通路的HRAS、KRAS、NRAS及RET是PTC中的高频突变基因。有2例病例分别含35个和40个突变基因,存在很大肿瘤负荷,结合临床数据发现均为术后复发患者。常见的BRAF V600E突变并非都是体细胞突变（64%）,也可为胚系突变（29%）;用NGS和qPCR同时检测验证V600E时,2种方法检出结果的相符率达80%。结论： CD44和CCND2基因在乳头状甲状腺癌癌组织中高表达,BRAF、RAS、FGFRs、KIT和MYC等基因的肿瘤特异性突变可能作为临床上对PTC患者实施个体化治疗所需的分子靶点。NGS和qPCR技术对BRAF V600E检测具有很高平行性,联合应用可提高检出率。

Objective: To identify the clinical molecular targets in papillary thyroid cancer (PTC) by analyzing the diffe-rential expression of the NF-κB signal pathway related genes and mutations of 52 solid tumor-related genes in PTC against normal adjacent tissue (NAT). Methods: RNA and DNA of tumor and NAT were extracted from formalin-fixed paraffin-embedded (FFPE) tissue of 20 papillary thyroid cases. The mRNA and protein expressions of NF-κB signal pathway related gene CD44, BCL2, CCND2, c-FLIP, IκBα, A20 and ABINs were detected by quantitative reverse transcription polymerase chain reaction (qRT-PCR) and immunohistochemistry, respectively. Mutations in 5 relapse cases were detected by targeted next-generation sequencing (NGS). Results: Expressions of CD44 and CCND2 were significantly higher at both RNA and protein level in PTC tumor tissue than those in NAT. The expressions of NF-κB signal pathway related genes were not significantly different between PTC having and not having lymph node metastasis. Results of NGS showed that ALK, BRAF, FGFR3/4, KIT, MYC and MAPK signal pathway genes HRAS, KRAS, NRAS, RET were mutated in PTC cases with a high frequency. Two cases had 35 and 40 mutation genes, respectively, and having high tumor burden; clinical data showed that these two were relapse cases after operation. BRAF V600E mutation was not always a tumor specific mutation (64%), but also could be a germline mutation (29%). Real-time quantitative PCR (qPCR) and NGS had a high consistent rate (80%) for the detection of V600E mutation. Conclusions: The high expressions of CD44 and CCND2 in PTC tumor tissue and the tumor specific mutations in BRAF, RAS, FGFRs, KIT and MYC genes might be used as clinical molecular markers for selection of targeted therapy of individual PTC patient. NGS and qPCR had high consistence for detection of BRAF V600E, and used jointly could improve the detection rate.