环氧化酶-2在分化型甲状腺癌中的表达及意义

目的探讨环氧化酶-2（COX-2）在甲状腺乳头状癌与滤泡癌中的表达及其临床意义。方法采用免疫组织化学SP法,检测36例甲状腺乳头状癌和15例甲状腺滤泡癌中COX-2的表达。结果 COX-2表达于癌细胞的胞质,其阳性表达率乳头状癌组（72.22%）高于滤泡癌组（40.00%）,有淋巴结转移组（78.57%）高于无淋巴结转移组（43.48%）,差异均有统计学意义,COX-2表达还与肿瘤TNM分期有关。结论 COX-2对分化型甲状腺癌的转移起重要作用,其具体机制还有待进一步研究。     

Gene expression profiling associated with the progression to poorly differentiated thyroid carcinomas

Background:

Poorly differentiated thyroid carcinomas (PDTC) represent a heterogeneous, aggressive entity, presenting features that suggest a progression from well-differentiated carcinomas. To elucidate the mechanisms underlying such progression and identify novel therapeutic targets, we assessed the genome-wide expression in normal and tumour thyroid tissues.

Methods:

Microarray analyses of 24 thyroid carcinomas – 7 classic papillary, 8 follicular variants of papillary (fvPTC), 4 follicular (FTC) and 5 PDTC – were performed and correlated with RAS, BRAF, RET/PTC and PAX8-PPARG alterations. Selected genes were validated by quantitative RT–PCR in an independent set of 28 thyroid tumours.

Results:

Unsupervised analyses showed that gene expression similarity was higher between PDTC and fvPTC, particularly for tumours harbouring RAS mutations. Poorly differentiated thyroid carcinomas presented molecular signatures related to cell proliferation, poor prognosis, spindle assembly checkpoint and cell adhesion. Compared with normal tissues, PTC had 307 out of 494 (60%) genes over-expressed, FTC had 137 out of 171 (80%) genes under-expressed, whereas PDTC had 92 out of 107 (86%) genes under-expressed, suggesting that gene downregulation is involved in tumour dedifferentiation. Significant UHRF1 and ITIH5 deregulated gene expression in PDTC, relatively to normal tissues, was confirmed by quantitative RT–PCR.

Conclusion:

Our findings suggest that fvPTC are possible precursors of PDTC. Furthermore, UHRF1 and ITIH5 have a potential therapeutic/prognostic value for aggressive thyroid tumours.     

The Role of Claudin-1 Expression in Follicular and Papillary Thyroid Neoplasm

Objective: This study evaluated differences in Claudin-1 expression between follicular adenoma (FA), follicular thyroid carcinoma (FTC), follicular variant papillary thyroid carcinoma (FV-PTC), and papillary thyroid carcinoma (PTC). Material and methods: This study used a cross-sectional approach. Immunostaining using the polyclonal antibody Claudin-1 was performed on 75 samples divided into 20 samples for follicular adenoma, follicular thyroid carcinoma, papillary carcinoma, and 15 samples of follicular variant thyroid carcinoma, respectively. Results: Claudin-1 expression is detected on the cytoplasmic membrane of tumor cells and appears to be varied among thyroid neoplasms. The claudin-1 expression score revealed a statistically significant difference between FA against FV-PTC, FA versus (vs) PTC, and FTC vs PTC, with median values of 4 vs 6 (p = 0.016), 4 vs 8 (p = 0.001), and 5 vs 8 (p = 0.002), respectively. However, there was no statistically significant difference in scores between the FA and the FTC (4 vs 5), or between the FTC and the FV-PTC groups (5 vs 6 (p=1,000). Conclusion: These results suggest that Claudin-1 may be capable of discriminating follicular adenoma from classic and follicular variant of papillary thyroid carcinoma. It can also differentiate follicular thyroid carcinoma and papillary thyroid carcinoma, especially for cases challenging to assess by hematoxylin and eosin staining. It still holds promise in providing targeted cancer therapy.     

细胞角蛋白-19 波形蛋白血管内皮生长因子-C和环氧化物酶-2 在甲状腺乳头状癌和滤泡癌中的表达及意义

目的:探讨细胞角蛋白-19（CK- 19）、波形蛋白（Vimentin）、血管内皮生长因子-C（VEGF-C）和环氧化物酶-2（COX-2）表达与甲状腺乳头状癌和滤泡癌分化、浸润、转移的关系。方法:采用免疫组织化学SP法检测94例乳头状癌和54例滤泡癌中CK- 19、Vimentin、VEGF-C、COX-2 的表达。结果:CK- 19、VEGF-C、COX-2 表达于癌细胞的胞浆,Vimentin 在癌组织的间质和癌细胞的胞浆均有表达,其表达差异从阳性率和阳性强度两个方面予以统计分析。乳头状癌中CK- 19表达的阳性率（90.4%）高于滤泡癌（61.1%）,Vimentin、VEGF-C 和COX-2 表达的阳性率（100% 、86.2% 、81.9%）近似于滤泡癌（100% 、87.0% 、81.5%）。 乳头状癌无转移组和有转移组（含原发灶和转移灶）四种指标的阳性率均相近,滤泡癌高分化型组与低分化型组相比,除Vimentin 的阳性率均为100% 外,其他三种指标CK- 19、VEGF-C、COX-2 均为高分化型组阳性率（74.1% 、88.9% 和85.2%）,高于低分化型组阳性率（48.1%、85.2%和77.8%）。 Vimentin 和VEGF-C 的免疫阳性表达强度在乳头状癌中高于滤泡癌,CK- 19和COX-2 在乳头状癌和滤泡癌中的表达相近。四种免疫学指标的免疫阳性强度在乳头状癌原发灶、转移灶、滤泡癌高分化组、低分化型组间均无明显差异。乳头状癌中CK- 19与Vimentin 间免疫表达强度无相关性,VEGF-C 和COX-2 间呈正相关关系。结论:综合检测CK- 19、Vimentin、VEGF-C 和COX-2 四种免疫学指标,对探讨甲状腺乳头状癌和滤泡癌分化、浸润、转移和预后评估具有一定的参考价值。      

Expression of telomerase genes in thyroid carcinoma

Telomerase (T) is a ribonucleoprotein complex that includes the telomerase RNA component (hTR), telomerase associated protein (TP1) and the telomerase catalytic subunit (hTERT). Telomerase has been shown in stem cells and found to be activated in tumor tissues and immortalized cells. We wanted to test whether the expression of the telomerase complex subunits correlate with the enzyme activity in human thyroid tissue. Hence, we determined the expression of hTERT, hTR and TP1 mRNA by RT-PCR and compared the results to telomerase activity as detected by the telomeric repeat amplification protocol (TRAP) assay. Fifteen benign goiters (G), 11 follicular carcinomas (FTC) including 2 oncocytic follicular carcinomas (also called Hurthle cell carcinoma, oFTC), 12 papillary carcinomas (PTC) including 3 microcarcinomas (mPTC), and 12 undifferentiated anaplastic thyroid carcinomas (UTC) were investigated. Experienced pathologists performed histological and pTNM classification in each specimen. RT-PCR analysis revealed that TP1 was ubiquitously expressed in all G and carcinomas. hTR was expressed in 4 out of 15 G, in 2 out of 3 mPTC, in 5 out of 9 PTC, in 5 out of 9 FTC, in all oFTC and in 9 out of 12 UTC samples. Regarding all carcinomas, no statistically significant correlation was observed between hTR-expression and tumor stage, lymph node or distant metastasis. hTERT-expression was associated with malignancy and tumor stage. All mPTC and 13 out of 15 G did not express hTERT, whereas all samples of pT3-4 tumor stage of FTC, PTC, UTC and all oFTC were positive for hTERT. No telomerase activity could be detected in G. Telomerase activity in carcinoma was only measurable in tissues that expressed the catalytic subunit hTERT. Our data indicate that telomerase activity is up-regulated in neoplastic cells. In contrast to TP1 and hTR, hTERT and telomerase activity may be of help in identifying invasive tumors and may be additional markers for classification of benign goiter and malignant thyroid carcinoma.     

甲状腺乳头状癌BRAF基因突变及表达的临床意义研究

目的探讨BRAF基因点突变及B-raf蛋白表达在甲状腺乳头状癌发生中的临床意义。方法应用聚合酶链式反应(PCR)技术检测65例甲状腺病变石蜡组织中BRAF点突变,应用免疫组化方法检测112例甲状腺病变组织中B-raf蛋白的表达情况,并比较BRAF基因突变和B-raf蛋白表达的相关性。结果在46例甲状腺乳头状癌中有21例发生BRAF的点突变,突变率为45.7。BRAF基因突变位于第15外显子的1799位点,胸腺嘧啶突变为腺嘌呤(T1799A)。在结节性甲状腺肿和滤泡状癌中未检测到BRAF的突变。乳头状癌BRAF基因突变率与结节性甲状腺肿比较,差异具有统计学意义(P<0.05)。但是与患者的性别、年龄、组织学类型、淋巴结转移和肿瘤分期无相关性(P>0.05)。在乳头状癌、滤泡状癌和结节性甲状腺肿中B-raf蛋白表达阳性率分别为65.1、47.6和15.4。结果显示,乳头状癌B-raf蛋白阳性表达率与良性病变比较,差异具有统计学意义(P<0.05)。在乳头状癌中BRAF基因突变与B-raf蛋白表达水平呈正相关(P<0.05)。乳头状癌与滤泡状癌比较,B-raf蛋白表达水平两组间无统计学意义(P>0.05)。结论甲状腺乳头状癌BRAF基因突变率和蛋白表达水平的增高,提示BRAF基因在乳头状癌发病中可能发挥重要的作用。对甲状腺肿瘤的病理诊断也具有辅助价值。     

山东省沿海与内陆甲状腺癌患病模式变迁

目的：观察沿海城市（青岛市）和内陆城市（临沂市）11年来甲状腺乳头状癌（PTC）、滤泡型癌（FTC）、髓样癌（MTC）、未分化癌（ATC）患病率的变化趋势,并将内陆城市与沿海城市进行比较,以探讨碘营养与甲状腺癌的关系。方法：根据国际肿瘤分类标准,利用石蜡切片等方法对青岛大学医学院附属医院病理科、临沂市人民医院病理科和临沂市第二人民医院病理科1998年-2008年间收检的甲状腺恶性肿瘤进行重新分类,回顾性分析患者的平均确诊年龄、性别比例、构成比。结果：11年来4类甲状腺恶性肿瘤沿海城市（青岛）的总例数为734例,其中PTC占85.56%、FTC占8.86%、MTC占4.36%、ATC占1.22%。4类中女性发病均高于男性（男女比例1∶1.25-3.94）。PTC的平均确诊年龄最低（45.39岁）,ATC最高（61.56岁）。内陆城市（临沂）的总例数为421例,其中PTC占82.90%、FTC占9.74%、MTC占4.75%、ATC占2.61%。4类甲状腺癌中女性发病均高于男性（男女比例1∶2.33-7.20）。PTC的平均确诊年龄最低（42.09岁）,ATC最高（67.91岁）。沿海与内陆的PTC均呈逐年递增趋势,FTC、MTC和ATC递增趋势不明显。结论：11年来沿海与内陆的PTC均呈逐年递增趋势,FTC、MTC和ATC递增趋势不明显。内陆PTC患病年龄低于沿海。碘营养等因素可能参与PTC的发生,其确切机制值得进一步研究。     

High Expression of Claudin-1 Protein in Papillary Thyroid Tumor and its Regional Lymph Node Metastasis

Claudins, known as major contributors in the formation of the tight junction, are differentially expressed in malignant tumors as compared to the corresponding healthy tissues. Therefore, they are thought to play a role in carcinogenesis and tumor progression. Altered expression of claudin-1 has been reported in several tumor types including endometrial, papillary renal cell and colonic carcinoma, and increased claudin-1 mRNA levels have been observed in papillary thyroid carcinoma (PTC). In this study, we aimed at determining the pattern of claudin-1 expression in various types of thyroid lesions at the protein level and investigating the immunolocalization of β-catenin reported to regulate claudin-1 expression. Samples included 19 PTCs, ten cases of corresponding regional lymph node metastasis, eight papillary microcarcinomas (PMC), 17 follicular thyroid carcinomas (FTC) and 19 follicular adenomas (FA). All cases were evaluated by quantitative immunohistochemistry. Conspicuous claudin-1 immunostaining was detected in the majority of PTC/PMC primary tumors and lymph node metastases (19/27 and 9/10, respectively). On the other hand, we found weak or no claudin-1 expression in any of the FA and FTC cases or peritumoral non-malignant thyroid tissues. Our data prove that high claudin-1 protein expression is specific for PTC and its regional lymph node metastases, while we failed to verify that claudin-1 is regulated by β-catenin in thyroid tumors. Based on these results, claudin-1 may be a useful tumor marker for PTC.     

CD57在滤泡状甲状腺肿瘤中的表达及其临床意义

目的　观察CD5 7在滤泡状甲状腺癌 (FTC)和滤泡状甲状腺腺瘤 (FTA)中的表达。方法　采用免疫组织化学SP法 ,共检测了91例甲状腺组织中CD5 7的表达 ,其中FTC 2 5例 ,FTA 5 0例 ,瘤旁正常甲状腺组织 16例。结果　① 16例瘤旁正常的甲状腺组织无一例表达CD5 7。②CD5 7在FTC、FTA上的表达率分别为 88 0 0 % ( 2 2 /2 5 )、2 8 0 0 ( 14 /5 0 )。CD5 7在FTC与FTA间的阳性表达均有显著性差异 (P <0 0 0 1)。结论　CD5 7可作为鉴别FTC与FTA较好的肿瘤标志物     

Stratification of follicular thyroid tumours using data‐independent acquisition proteomics and a comprehensive thyroid tissue spectral library

Thyroid nodules occur in about 60% of the population. A major challenge in thyroid nodule diagnosis is to distinguish between follicular adenoma (FA) and carcinoma (FTC). Here, we present a comprehensive thyroid spectral library covering five types of thyroid tissues. This library includes 121 960 peptides and 9941 protein groups. This spectral library can be used to quantify up to 7863 proteins from thyroid tissues, and can also be used to develop parallel reaction monitoring (PRM) assays for targeted protein quantification. Next, to stratify follicular thyroid tumours, we compared the proteomes of 24 FA and 22 FTC samples, and identified 204 differentially expressed proteins (DEPs). Our data suggest altered ferroptosis pathways in malignant follicular carcinoma. In all, 31 selected proteins effectively distinguished follicular tumours. Of those DEPs, nine proteins were further verified by PRM in an independent cohort of 18 FA and 19 FTC. Together, we present a comprehensive spectral library for DIA and targeted proteomics analysis of thyroid tissue specimens, and identified nine proteins that could potentially distinguish FA and FTC.     

Differential miRNA expression profiles in variants of papillary thyroid carcinoma and encapsulated follicular thyroid tumours

Background:

Recent studies showed a significant upregulation of distinct microRNAs (miRNAs) in papillary thyroid carcinoma (PTC). The objective of this study was to explore whether this upregulation could also be assigned to distinct histomorphological variants of PTC, especially the follicular variant and other encapsulated follicular thyroid tumours.

Methods:

We used total RNA of 113 formalin-fixed paraffin-embedded tissues of 50 PTCs ((10 conventional type (PTC-CT), 10 tall cell variants (PTC-TCVs), 30 follicular variants (PTC-FVs)), 10 follicular adenomas (FAs), 10 multinodular goitres (MNGs), 21 follicular thyroid carcinomas and 22 well-differentiated tumours of unknown malignant potential (WDT-UMP) to analyse the miRNA expression pattern of five selected miRNAs (146b, 181b, 21, 221 and 222) using RT–PCR TaqMan miRNA assay to explore the diagnostic utility of this method.

Results:

The mean values of the expression pattern of all miRNAS in PTCs show a statistically significant difference from those in MNG and FA with fold changes up to 90 for miRNA 146b, P<0.001. No differences in expression pattern could be showed between MNG and FA. The PTC-FVs differ significantly from FA in all five miRNAS, from MNG in three and from WDT-UMP in one miRNA with fold changes between 1.7 and 21.2, but failed to be of diagnostic value regarding individual cases with substantial overlaps.

Conclusion:

We conclude that analysis of a set of five selected miRNAS distinguish common variants of PTC from FA/MNG but failed to be a useful diagnostic method in individual and doubtful cases, especially in the differential diagnosis of encapsulated follicular thyroid tumours.     

甲状腺滤泡性肿瘤形态定量分析

本文应用MIAS-300型图像分析系统对甲状腺良、恶性滤泡性肿瘤各30例及正常甲状腺组织8例进行了形态定量测定及比较分析,旨在探寻形态定量指标在鉴别诊断中的价值,并建立数据库,为计算机协助诊断疾病打下基础。常规制片,用个体测量法及群体测量法分别分析了每例100～145个滤泡上皮细胞,获得6项参数,包括核面积、核周长、核形状因子、等效核直径、核黑度及核质比。经统计学分析,显示上述各项参数在良恶之间比较均有高度显著性差异,表明这些参数在甲状腺滤泡性肿瘤良恶鉴别中有一定价值,为数学病理学研究提供了一定资料。