Galectin-3、CK19和TPO表达在甲状腺良恶性肿瘤病理诊断中的价值

目的比较甲状腺病变中galectin-3、CK19和甲状腺过氧化物酶（thyroid peroxidase,TPO）蛋白表达的差异,寻找有助于诊断良恶性肿瘤的标志物。方法采用免疫组化EnVision二步法检测134例甲状腺癌（123例乳头状癌、11例滤泡癌）、34例甲状腺腺瘤、20例结节性甲状腺肿和10例桥本甲状腺炎中galectin-3、CK19和TPO蛋白的表达。结果Galectin-3、CK19和TPO在甲状腺乳头状癌（包括主要的3个亚型）与良性病变中的表达差异有显著性（P〈0．01）,在乳头状癌各亚型间和各良性病变之间的表达差异均无显著性（P〉0．05）。滤泡癌和良性病变中galectin-3表达差异有显著性（P〈0．01）。滤泡癌和腺瘤中CK19、TPO的表达差异有显著性（P〈0．05,P〈0．01）。结论甲状腺恶性肿瘤特别是乳头状癌中galectin-3和CK19的表达有增高,而TPO表达缺失。3种蛋白分别是诊断甲状腺肿瘤特别是乳头状癌的有用标志物,联合使用结果更可靠。     

甲状腺乳头状癌CD151和CK19mRNA表达及意义

目的探讨甲状腺乳头状癌中CD151和CK19 mRNA表达及其意义。方法采用RT—PCR方法检测54例甲状腺的良恶性病变组织中CD151和CK19 mRNA的表达,并与临床及病理资料进行分析比较。结果CD151和CK19 mRNA在甲状腺乳头状癌组中的表达与滤泡性腺瘤组、结节性甲状腺肿组以及正常甲状腺组比较均差异具有显著性（P〈0．01）;CK19 mRNA表达与甲状腺癌淋巴结转移有关（P〈0．05）;在甲状腺乳头状癌中,CD151和CK19基因mRNA的表达呈正相关（P〈0．01）。结论CD151和CK19基因共同参与甲状腺癌的发生、发展,并对淋巴结转移的判断有意义。     

CK19、CK20在甲状腺乳头状癌诊断中的应用价值

目的：探讨甲状腺癌中CK19、CK20蛋白的表达，提高甲状腺的诊断与鉴别诊断水平。方法：应用免疫组化染色对70例甲状腺癌（15例经典型乳头状癌、34例滤泡型乳头状癌、3例Warthin乳头状癌、2例透明细胞型乳头状癌、例柱状细胞型乳头癌、15例滤泡性癌），10例甲状腺腺瘤、10例结节性甲状腺肿和5例标本甲状腺炎中CK19、CK20的表达进行观察。结果:CK19在甲状腺疾病中的表达：55例乳头状癌中，53例为中、强阳性，2例为弱阳性；15例滤泡性癌中，13例为阴性、弱阳性，2例为中、强阳性，两者之间差异存在显著性（P＜0．05）。各癌旁滤泡、10例滤泡性腺瘤、10例结节性甲状腺肿的滤泡、5例桥本甲状腺炎也主要为阴性、弱阳性，个别为中等阳性。对CK20的表达，各型甲状腺乳头状癌、滤泡性癌、癌旁滤泡及滤泡状癌和乳头状增生、多灶性分布的甲状腺泡型乳头状癌和各种滤泡性病变有帮助，可提高甲状腺良恶性病变诊断的准确率及鉴别诊断水平。CK20对鉴别诊断的帮助不大。     

细胞角蛋白和Ret蛋白在甲状腺乳头状癌中的表达

目的　探讨甲状腺乳头状癌中细胞角蛋白 (CK)和ret的表达及其在病理诊断中的价值。方法　分别采用免疫组织化学EnVision法及LSAB法检测CK19、CK17、CK8、CK2 0及ret在 6 9例甲状腺乳头状癌、14例结节性甲状腺肿和 4 2例癌旁正常滤泡中的表达。结果　CK19、ret在乳头状癌组织的阳性率 (85 5 %、6 8 1% )明显高于结节性甲状腺肿和正常滤泡组织阳性率 (2 5 0 %、5 4 % ) ,二者差异有统计学意义 (P <0 0 1)。CK17在乳头状癌中有少量表达 (15 9% ) ,主要集中在鳞化及分化差或小灶浸润区域。分别有 75 4 %及 2 6 8%的乳头状癌及良性区域CK8染色阳性 ,所有病例CK2 0阴性。结论　CK19、CK17及ret在甲状腺乳头状癌中表达增加 ,在病理诊断中有一定价值。     

甲状腺微小乳头状癌中Galectin-3、CK19、HBME-1和CD56的表达及意义

目的探讨结节性甲状腺肿合并甲状腺微小乳头状癌中Galectin-3、CK19、HBME-1及CD56的表达及意义。方法采用免疫组化SP法检测10例结节性甲状腺肿合并甲状腺微小乳头状癌中Galectin-3、CK19、HBME-1和CD56的表达水平。结果 Galectin-3、CK19和HBME-1在甲状腺微小乳头状癌中均呈中至强阳性表达,而在结节性甲状腺肿中主要呈阴性或弱阳性表达。然而在10例甲状腺微小乳头状癌中有8例CD56的表达均为阴性,2例呈轻度阳性着色;周围结节性甲状腺肿组织9例均呈中至强阳性表达,仅1例呈轻度阳性着色。结论 Galectin-3、CK19、HBME-1及CD56联合检测将进一步提高甲状腺微小乳头状癌的准确性。     

Bcl-2和ER在甲状腺乳头状癌中的表达及意义

目的：探讨细胞凋亡相关基因bcl-2和雌激素受体（ER）在甲状腺乳头状癌中的表达及意义。方法：用免疫组织化学sP法研究不同甲状腺组织中ER和bcl-2的表达，其中甲状腺乳头状癌50例、甲状腺良性腺瘤30例、腺瘤旁正常组织16例。结果：甲状腺乳头状癌组织中ER和bcl-2蛋白阳性率分别为54．0％（27／50）、66．0％（33／50），甲状腺良性腺瘤中ER和bcl-2蛋白阳性率分别为26．7％（8／30）、36．7％（11／30），正常甲状腺组织中ER阳性率为12．5％（2／16）、而bcl-2则不表达。甲状腺乳头状癌中ER的表达明显高于甲状腺良性腺瘤和正常甲状腺组织（P〈0．05），bcl-2的表达明显高于甲状腺良性腺瘤（P〈0．05）；且甲状腺乳头状癌中bcl-2和ER表达存在正相关关系（P〈0．05）。结论：雌激素在甲状腺乳头状癌的发生、发展中有促进细胞增殖的作用；对ER和bcl-2的联合检测在甲状腺乳头状癌的诊断及内分泌治疗有一定的临床意义。     

细胞角蛋白19、galectin-3、HBME-1在甲状腺病变上的表达及鉴别诊断意义

目的 研究细胞角蛋白(CK)19、galectin(Gal)-3、HBME-1在甲状腺不同病变表达的特点及鉴别诊断中的应用价值。方法 应用免疫组织化学EnVision法检测了21例结节性甲状腺肿(结甲)、14例毒性甲状腺肿(甲亢)、15例甲状腺滤泡性腺瘤(腺瘤)、13例滤泡性癌、13例滤泡型乳头状癌及48例经典型乳头状癌中单克隆抗体CK19、Gal-3、HBME-1的表达。结果 甲状腺病变中3种标记表达均位于细胞质;CK19、Gal-3、HBME-1的表达在甲状腺良性病变(结甲、甲亢、腺瘤)大多为弱阳性或阴性,而滤泡性癌阳性明显增加、乳头状癌(滤泡型及经典型)大多为中、强阳性,3种标记在甲状腺不同病变的阳性表达率结甲为52．4％(11／21)、9．5％(2／21)、19．0％(4／21),甲亢为50．0％(7／14)、7．1％(1／14)、7．1％(1／14),腺瘤为60％(9／15)、13．3％(2／15)、13．3％(2／15),滤泡性癌为76．9％(10／13)、61．5％(8／13)、53．8％(7／13),滤泡型乳头状癌为：100％(13／13)、84．6％(11／13)、92．3％(12／13),经典型乳头状癌为100％(48／48)、93．8％(45／48)、95．8％(46／48);在甲状腺良性病变(结甲、甲亢、腺瘤)与恶性病变(滤泡性癌、乳头状癌)间3种标记差异均有显著性(P均=0．000);同时3种标记在滤泡样病变即腺瘤、滤泡性癌和滤泡型乳头状癌间亦有显著差异(CK19：P=0．038,Gal-3：P=0．001,HBME-1：P=0．000)。结甲有9例,甲亢有7例,腺瘤有6例3种标记均不表达,滤泡性癌仅有1例,而乳头状癌(滤泡型及经典型)没有病例3种标记均不表达,同一病例有2种以上阳性表达在结甲、甲亢、腺瘤、滤泡性癌、滤泡型乳头状癌和经典型乳头状癌中分别为14．2％(3／21)、21．4％(3／14)、20．0％(3／15)、69．2％(9／13)、92．3％(12／13)、100．0%(48／48),在甲状腺良性病变与恶性病变间以及滤泡样病变间差异亦有显著性(P=0．000)。结论 CK19、Gal-3、HBME-1的检测尤其是联合检测对甲状腺病变的诊断、鉴别诊断具有较高的实用价值。     

甲状腺乳头状癌中AEG-1、Livin的表达及临床意义

目的检测星形细胞上调基因-1 (astrocyte elecated gene-1,AEG-1)和凋亡抑制蛋白(Livin)在甲状腺乳头状癌组织中的表达及其相关性,为甲状腺乳头状癌提供新的诊断参考指标和治疗靶向位点。方法采用免疫组化SP法检测92例甲状腺乳头状癌组织、45例结节性甲状腺肿组织及60例同期手术的癌旁正常甲状腺组织中AEG-1和Livin的表达,并分析其表达水平与临床病理特征的关系。结果AEG-1与Livin在甲状腺乳头状癌组织中的阳性率显著高于结节性甲状腺肿与癌旁正常甲状腺组织(P 0. 05),但两者在结节性甲状腺肿组织与癌旁正常甲状腺组织中的表达,差异无显著性(P 0. 05)。AEG-1蛋白表达水平与患者肿瘤的临床分期、淋巴结转移、腺外侵犯均密切相关(P 0. 05);Livin蛋白表达水平与患者肿瘤的临床分期、淋巴结转移、TG-Ab值、TPO-Ab值有相关性(P 0. 05)。甲状腺乳头状癌组织中AEG-1与Livin蛋白表达水平呈显著的正相关(rs=0. 673,P 0. 001)。结论 AEG-1、Livin与甲状腺乳头状癌的侵袭和转移关系密切,在临床中联合检测两者将更有利于甲状腺乳头状癌的早期诊断和判断患者的预后,有望为靶向治疗提供新的靶点。     

甲状腺乳头状癌中CK19、Galectin-3、HBME-1和TPO的表达及意义

目的 探讨CK19、Galectin-3、HBME-1和TPO在甲状腺良恶性病变中的表达及联合应用在甲状腺乳头状癌鉴别诊断中的价值.方法 采用免疫组化EnVision法检测68例甲状腺乳头状癌、31例甲状腺腺瘤、19例结节性甲状腺肿和15例桥本甲状腺炎中CK19、Galectin-3、HBME-1和TPO的表达.结果 在甲状腺乳头状癌中CK19、Galectin-3和HBME-1的阳性表达率分别为98.5%、98.5%、80.9%,TPO的阴性表达率为89.7%;在甲状腺良性病变中CK19、Galectin-3和HBME-1阳性表达率分别是24.6%、21.5%、1.5%,TPO的阴性表达率是1.5%.CK19、Galectin-3、HBME-1和TPO在甲状腺乳头状癌与良性病变中的表达差异有显著性(P<0.001).联合应用四种抗体在鉴别甲状腺乳头状癌与良性病变时的敏感性、特异性、准确度分别为95.6%、98.5%、97%.结论 CK19、Galectin-3、HBME-1和TPO是诊断甲状腺乳头状癌的重要标志物.联合应用四种抗体在甲状腺乳头状癌与良性病变的鉴别诊断中具有重要价值.     

能谱CT测定甲状腺组织碘含量的临床研究

目的利用宝石cT能谱成像技术（GSI）测量甲状腺乳头状癌与结节性甲状腺肿患者非瘤区甲状腺组织内碘含量,并与正常甲状腺内碘含量比较,探索甲状腺碘含量与甲状腺疾病的关系。方法收集北京大学深圳医院2011年8月—2013年7月接受甲状腺能谱CT检查并经手术病理证实的甲状腺病变患者36例,其中甲状腺乳头状癌18例、结节性甲状腺肿18例。正常甲状腺对照组15例,为怀疑颈部或颈椎疾病进行能谱CT扫描,无甲状腺功能及结构异常。使用GE公司宝石CT（GEDiscoveryCT750HI）CT）GSI模式扫描,图像采集后用GSIViewer浏览器处理,在碘基图中对结节性甲状腺肿与甲状腺乳头状癌病变外正常甲状腺组织及正常对照组甲状腺组织进行测量,采用Wilcoxon秩和检验及t检验进行统计学分析并探究其碘含量的分布特点。结果甲状腺乳头状癌中非瘤区腺体组织与正常甲状腺、结节性甲状腺肿中非瘤区腺体组织与正常甲状腺碘含量两两比较,差异均有统计学意义（P〈0．05）,甲状腺乳头状癌中非瘤区腺体组织与结节性甲状腺肿中非瘤区腺体组织的碘含量比较,差异无统计学意义（P〉0．05）,甲状腺乳头状癌中非瘤区腺体组织与结节性甲状腺肿中非瘤区腺体组织甲状腺碘含量呈“U”型分布。结论cT能谱成像技术作为一种新的方法,能够无创性地检测人体甲状腺碘含量,此技术将有助于进一步揭示甲状腺碘含量与甲状腺疾病关系。     

Overexpression of cytokeratin 17 is associated with the development of papillary thyroid carcinoma and the presence of lymph node metastasis

Cytokeratin 17 (CK17), a basal/myoepithelial cell keratin, appears to play an important role in the progression of several human malignancies. Increased CK17 expression has previously described in cases of papillary thyroid carcinoma (PTC). However, no studies to date have investigated the clinical significance of CK17 expression in patients with PTC. The aim of this study was to compare the expression of CK17 in patients with PTC with that observed in normal thyroid tissue and benign thyroid lesions, and to examine the relationship between CK17 expression and clinicopathologic characteristics of patients with PTC. CK17 protein expression was evaluated by immunohistochemistry on tissue microarrays containing thyroid tissue samples from 108 PTCs, 16 nodular goiters, and 81 healthy controls. Sixty-five of the 108 (60.2%) PTC tissue samples exhibited positive CK17 expression, whereas all nodular goiters and normal thyroid tissue samples showed a complete absence of CK17 immunoreactivity. The difference in frequency of CK17 positivity between PTC (65/108, 60.2%), normal thyroid tissue (0/81, 0.0%), and benign thyroid lesions (0/16, 0.0%) was statistically significant (P<0.001). Positive CK17 expression in PTC was significantly associated with the presence of lymph node metastasis (P=0.024) and higher pN stage (P=0.028). Expression of CK17 is significantly increased in cases of PTC compared to normal tissue and benign thyroid lesions, and CK17 overexpression is associated with the presence of lymph node metastasis in patients with PTC. These findings suggest that CK17 is involved in the development and metastasis of PTC.     

RET/PTC and CK19 expression in papillary thyroid carcinoma and its clinicopathologic correlation

Recently, the rearrangement of RET proto-oncogene has been reported to be the most common genetic change in papillary thyroid carcinoma (PTC). However, its prevalence has been reported variably and its relation to clinical outcome has been controversial. The characteristic nuclear features of PTC usually render the diagnosis, but problem arises with equivocal cytologic features that are present focally. Although there remains some controversy, CK19 has been reported to be a useful ancillary tool for diagnosis of PTC. To evaluate the expression rate of RET/PTC rearrangement and CK19 in PTCs in a Korean population, we studied 115 papillary thyroid carcinomas in 3 mm-core tissue microarray based immunohistochemical analysis. The prevalence of Ret protein expression was 62.6% and the CK19 immunoreactivity was 80.9%. There was no statistically significant association between the Ret positivity and CK19 immunoreactivity, although the percent agreement of the two was relatively high. The clinicopathological variables did not correlate with the expression of Ret. In conclusion, the prevalence of Ret protein expression and its clinicopathological implications in a Korean population are not much different from those reported in previous studies. However, its detection via immunohistochemistry can be a useful diagnostic tool for diagnosing papillary thyroid carcinoma in conjunction with CK19.     

野生型RET和RET/PTC融合基因在成人散发性甲状腺乳头状癌中的表达及其意义

目的探讨野生型BET（WT-RET）及RET／PTC1、3融合基因在成人散发性甲状腺乳头状癌（PTC）中的表达及其与临床病理学指标的关系和意义。方法用逆转录-聚合酶链反应（RT-PCR）检测102例石蜡与新鲜（43例）甲状腺病变组织（PTC66例,对照组各种良恶性肿瘤及良性病变共36例）中WT-RET和RET／PTC1、3融合基因的表达并结合临床资料进行分析。结果（1）62％（41／66）PTC患者≥40岁。38％（25／66）PTC伴淋巴细胞性甲状腺炎,59％（39／66）伴淋巴结转移,5例（7.6％）有远处转移。（2）RET原癌基因的酪氨酸激酶区（BET-TK）检出率为68．1％（45／66）。BET原癌基因断裂点（BP）与TK的同时检出率在PTC中28．8％（19／66）,腺瘤中12．5％（1／8）,表明存在WT-BET转录物。（3）RET／PTC检出率21．2％（14／66）,其中5例BET／PTC1阳性（7．6％）,9例RET／PTC3阳性（13．6％）。6例（9％）PTC同时表达BET／PTC和WT-BET。36例对照组病例中未检测到RET／PTC融合基因。（4）统计学分析,PTC病例中WT-BET与RET／PTC1融合基因的表达与性别、年龄、肿瘤大小、多灶性、伴淋巴细胞浸润及淋巴结转移等临床病理学指标无关（P〉0．05）。结论RET／PTC融合基因在散发性成人PTC中表达率低,其诊断和判断预后的价值不大。WT-BET在甲状腺肿瘤的滤泡形成过程中起一定作用。     

Overexpression of wild-type c-RET and zero prevalence of RET/PTC rearrangements are associated with papillary thyroid cancer (PTC) in Kuwait

Background

Papillary thyroid carcinoma (PTC) is common in Kuwait. The activation of the RET oncogene by DNA rearrangement (RET/PTC) is known to have an important role in PTC carcinogenesis. However, the real frequency of the RET/PTC expression in PTC is variable between different studies. This study seeks to determine the prevalence of RET/PTC and to analyze the RET oncogene expression associated with PTC in Kuwait.

Methods

RET expression and DNA rearrangements (RET/PTC 1, RET/PTC 2 and RET/PTC 3) were studied by RT–PCR in different thyroid diseases. Results were confirmed by the Southern blot and by immunohistochemistry. Quantitative real-time PCR was used to determine the level of RET mRNA expression in PTCs.

Results

Wild-type (nonrearranged) c-RET oncogene was overexpressed in 60% of PTC cases and absent in follicular thyroid carcinoma (FTC), anaplastic thyroid carcinoma (ATC), follicular adenomas (FA) or normal thyroid. No RET/PTC rearrangement was detected in any sample. The c-RET expression in Hashimoto's thyroiditis and multinodular goiter was limited to follicular cells with PTC-like nuclear changes.

Conclusions

The overexpression of wild-type c-RET is a characteristic molecular event of PTCs in Kuwait. The prevalence of RET/PTC is zero and among the lowest recorded in the world.     

Cytokeratin 19 immunoreactivity in the diagnosis of papillary thyroid carcinoma: a note of caution.

To evaluate the expression of cytokeratin (CK) 19, we stained sections obtained from formalin-fixed, paraffin tissue blocks of 35 thyroid tumors (follicular adenoma [FA], 20; papillary thyroid carcinoma [PTC], 10 follicular variant [FV] and 5 usual type) and scored the extent of staining as follows: 1+ (<5% positively stained cells), 2+ (5%-25% positively stained cells), 3+ (25%-75% positively stained cells), and 4+ (>75% positively stained cells). All 15 PTCs (including 10 FV-PTCs) were CK19 positive: 14 were 4+ and 1 (FV-PTC) was 2+. All 20 FAs also were CK19 positive: 15 were 1+, 1 was 2+, 4 were 3+, and none was 4+. In the FAs that were scored 1+, reactivity usually was confined to follicular cells lining cystically dilated atrophic follicles that lacked the typical nuclear features of PTC. The remaining FAs showed more diffuse reactivity, which was, however, less intense than that observed in the PTCs. Thus, immunoreactivity for CK19 is not specific for PTC, although we acknowledge that the extent and intensity of staining are considerably greater in this tumor than in FA. There were no significant differences in staining for CK19 between nonneoplastic follicles adjacent to PTCs and those adjacent to FAs.     

Minichromosome maintenance protein 3 is a candidate proliferation marker in papillary thyroid carcinoma

The proliferative capacity of tumor cells is a characteristic feature in the whole growing tumors. Many pathologists and clinicians have used the estimation of cell proliferation for prognostic information. Minichromosome maintenance protein 3 (MCM3) is known to have a role on the initiation and regulation of DNA replication during cell cycle. The aim of this study was to evaluate the potential applicability of one of the MCM proteins, MCM3, as a proliferation marker in papillary thyroid carcinoma (PTC) with correlation to clinicopathological parameters. We performed the immunohistochemical analysis for MCM3 and Ki-67 in 60 cases of PTC and Western blot analysis for MCM3 expression in 6 PTCs and normal thyroid tissues. The comparison of MCM3 labeling index (LI) to tumor size (P = 0.031) and extrathyroidal extension (P = 0.037) was statistically significant while that of Ki-67 LI to them was not. Moreover, a significant association was not observed between MCM3 and Ki-67, but the MCM3 LI was considerably higher. Western blot analyses revealed that the MCM3 protein expression levels were overexpressed in all PTCs. On the contrary, the levels of MCM3 were very low or absent in all normal thyroid tissues. Our results indicate that MCM3 may be a more reliable proliferation marker than Ki-67 in accessing the growth of tumor and evaluating tumor aggressiveness of PTC.     

Real-time quantitative RT-PCR identifies distinct c-RET, RET/PTC1 and RET/PTC3 expression patterns in papillary thyroid carcinoma

RET/PTC1 and RET/PTC3 are the markers for papillary thyroid carcinoma. Their reported prevalence varies broadly. Nonrearranged c-RET has also been detected in a variable proportion of papillary carcinomas. The published data suggest that a wide range in expression levels may contribute to the different frequency of c-RET and, particularly, of RET/PTC detection. However, quantitative expression analysis has never been systematically carried out. We have analyzed by real-time RT-PCR 25 papillary carcinoma and 12 normal thyroid samples for RET/PTC1, RET/PTC3 and for RET exons 10-11 and 12-13, which are adjacent to the rearrangement site. The variability in mRNA levels was marked and four carcinoma groups were identified: one lacking RET/PTC rearrangement with balanced RET exon levels similar to those of the normal samples (7/25 cases, 28%), the second (6/25 cases, 24%) with balanced RET expression and very low levels of RET/PTC1, the third with unbalanced RET exons 10-11 and 12-13 expression, high RET/PTC1 levels but no RET/PTC3 (7/25 cases, 28%), and the fourth with unbalanced RET expression, high RET/PTC1 levels and low levels of RET/PTC3 (5/25 cases, 20%). Papillary carcinomas with high RET/PTC1 expression showed an association trend for large tumor size (P=0.063). Our results indicate that the variability in c-RET and RET/PTC mRNA levels contributes to the apparent inconsistencies in their reported detection rates and should be taken into account not only for diagnostic purposes but also to better understand the role of c-RET activation in thyroid tumorigenesis.