Expression of galectin-3 in fine-needle aspirates as a diagnostic marker differentiating benign from malignant thyroid neoplasms.

BACKGROUND: Galectin-3 is a beta-galactoside-binding protein that has been reported to be expressed preferentially in thyroid malignancies. The current study was designed to substantiate this finding further and to establish a presurgical diagnostic modality of differentiating between benign and malignant thyroid neoplasms by analyzing galectin-3 expression in fine-needle aspirates. METHODS: The expression of galectin-3 was examined immunohistochemically in total of 172 specimens: 45 primary and 20 metastatic papillary carcinomas, 8 primary and 2 metastatic follicular carcinomas, 5 primary and 3 metastatic anaplastic carcinomas, 3 primary medullary carcinomas, 25 follicular adenomas, 3 goiters, and 58 adjacent normal thyroid tissue. Alternatively, epithelial cells were isolated from the fine- needle aspirates of 14 thyroid nodules and subjected to immunoblotting analysis of galectin-3. RESULTS: Immunohistochemical analysis revealed that all thyroid malignancies of follicular cell origin (including papillary, follicular, and anaplastic carcinomas) showed high and diffuse expression of galectin-3, whereas one of the three medullary carcinomas of parafollicular cell origin displayed weaker and focal expression of galectin-3. In contrast, neither benign thyroid adenomas, goiters, nor normal thyroid tissues expressed galectin-3. Immunoblot analysis of the isolated epithelial cells detected galectin-3 in nine thyroid nodules that were proven histologically to be malignant ( eight papillary carcinomas and one follicular carcinoma) after surgical intervention, whereas galectin-3 was not detected in five nodules proven to be benign follicular adenomas. CONCLUSIONS: Galectin-3 serves as a marker of thyroid malignancy of follicular cell origin. Analysis of galectin-3 expression in fine-needle aspirates enhances the differential diagnostic accuracy between benign and malignant thyroid neoplasms.     

血型抗原Lewis A和Lewis X在甲状腺乳头状癌中的表达

目的 :探讨血型抗原LewisA和LewisX在甲状腺乳头状癌中表达的意义。方法 :采用免疫组织化学EnVision方法 ,测定血型抗原LewisA和LewisX在 70例甲状腺乳头状癌和 70例癌旁正常甲状腺滤泡上皮 ,以及 16例甲状腺滤泡性腺瘤、17例结节性甲状腺肿和 14例桥本氏甲状腺炎中的表达。结果 :LewisA和LewisX在甲状腺乳头状癌中的表达阳性率分别为 94 3% ( 66/ 70 )和 85 7% ( 60 / 70 ) ,癌旁正常甲状腺滤泡上皮中的表达阳性率分别为 2 9% ( 2 / 70 )和 5 7% ( 4 / 70 )。肿瘤直径≥ 1cm组LewisX的表达强度较肿瘤直径 <1cm组明显增高 ,P <0 0 1。淋巴结转移组LewisX的表达强度较非转移组显著增高 ,P <0 0 1。结论 :LewisA和LewisX可作为诊断甲状腺乳头状癌的参考指标。LewisX的表达强度与肿瘤浸润及转移有关     

Nuclear Localization of Immunoreactive β-Catenin Is Specific to Familial Adenomatous Polyposis in Papillary Thyroid Carcinoma

Thyroid carcinoma is the first symptom in some patients with familial adenomatous polyposis (FAP). We evaluated the cellular localization of β-catenin in thyroid carcinomas associated (n=4) or not associated (n=173) with FAP, since loss of functional protein of the adenomatous polyposis coli (APC) gene leads to nuclear accumulation of β-catenin in adenomas and carcinomas of the FAP colon. Immunoreactive β-catenin was demonstrated at the cell membrane of glandular cells of the non-neoplastic thyroid and non-FAP carcinomas. On the other hand, cytoplasmic and nuclear accumulation of β-catenin is specific to FAP-associated papillary carcinomas. The abnormality in the APC/β-catenin pathway is thus also important in FAP-associated thyroid carcinoma, and β-catenin immunohistochemistry is a feasible screening method to identify occult FAP in young patients with thyroid tumors.     

Nuclear localization of immunoreactive beta-catenin is specific to familial adenomatous polyposis in papillary thyroid carcinoma.

Thyroid carcinoma is the first symptom in some patients with familial adenomatous polyposis (FAP). We evaluated the cellular localization of beta-catenin in thyroid carcinomas associated (n = 4) or not associated (n = 173) with FAP, since loss of functional protein of the adenomatous polyposis coli (APC) gene leads to nuclear accumulation of beta-catenin in adenomas and carcinomas of the FAP colon. Immunoreactive beta-catenin was demonstrated at the cell membrane of glandular cells of the non-neoplastic thyroid and non-FAP carcinomas. On the other hand, cytoplasmic and nuclear accumulation of beta-catenin is specific to FAP-associated papillary carcinomas. The abnormality in the APC / beta-catenin pathway is thus also important in FAP-associated thyroid carcinoma, and beta-catenin immunohistochemistry is a feasible screening method to identify occult FAP in young patients with thyroid tumors.     

Thyroid Carcinoma a Report of 206 Cases from an Area with Endemic Goitre

A retrospectiave analysis of 206 cases of thyroid carcinoma treated at a single surgical service in a region with endemic goitre is presented. In contrast to the marked female preponderance in surgically treated benign thyroid disorders, the frequency of thyroid carcinoma was almost equal in the two sexes (males/females = 100/106). The duration of symptoms was, as a rule, short and there was a predominance of advanced lesions (T3N3M1) even among the well-differentiated tumours. Papillary carcinoma was the most common histologic type (45%) but its proportion was considerably lower than usually reported from non-endemic regions. The average TSH level was significantly elevated in the carcinoma group and associated adenomatous changes were found in 40/100 cases which might suggest an etiologic role of increased TSH stimulation. Since most thyroid carcinomas had a very malignant clinical behaviour, surgical treatment aimed at near-total thyroidectomy which was performed in 62% of the cases. During a follow-up of up to eleven years recurrence in the remaining contralateral lobe occurred in 23% of patients with hemithyroidectomy and loco-regional recurrence was seen in 29% of patients with near-total thyroidectomy. Mortality was high in all histologic types, further underlining the aggressive biologic behaviour of thyroid cancers in an endemic area.     

Increased expression of insulin-like growth factor 1 (IGF-1) in multinodular non toxic goiter

Thyroid cell growth and function both in vivo and in vitro, are mainly regulated by TSH. Recent studies have shown that growth factors including insulin-like growth factors (IGF-1) have an important role in the control of thyrocyte proliferation and differentiation. The aim of this study was to evaluate the expression of the IGF-1 gene by Northern analysis and the IGF-1 tissue protein by radioimmunoassay in multinodular euthyroid goiters. The study population consisted of 20 patients with multinodular goiter (14 females and 6 males) living in a non endemic geographic area. All patients were euthyroid at the time of surgery and submitted to total or subtotal thyroidectomy. Samples of normal thyroid tissue were obtained from three patients who were operated due to laryngeal carcinomas. The IGF-1 protein content was increased in non toxic multinodular goiter, 22 ng/g vs. 14 in controls (p<0.03), as was IGF-1 gene expression (p<0.05). The increase in the steady state mRNA content correlated with the increase in the protein content (r=0.665; p<0.005). These results suggest that IGF-1 may play a role in proliferation events involved in benign hyperplastic thyroid diseases.     

甲状腺乳头状癌组织中DAPK1与RARβ的表达及临床意义

目的：通过检测新疆地区甲状腺乳头状癌组织及甲状腺良性病变组织中DAPK1和RARβ的蛋白表达特点,探讨其在甲状腺乳头状癌发生发展中的意义。方法：采用免疫组织化学S-P法,检测DAPK1与RARβ在新疆地区38例甲状腺乳头癌、21例甲状腺良性腺瘤、13例桥本氏甲状腺炎、6例结节性甲状腺肿和6例癌旁正常甲状腺组织中的蛋白表达,并分析其与临床病理因素间的关系。结果：DAPK1在甲状腺乳头状癌中的阳性表达明显低于甲状腺腺瘤、桥本氏甲状腺炎、结节性甲状腺肿和癌旁正常甲状腺组织（P〈0.01）;RARβ在甲状腺乳头状癌中的阳性表达明显高于在甲状腺良性腺瘤、桥本氏甲状腺炎、结节性甲状腺肿和癌旁正常甲状腺组织（P〈0.01）;DAPK1与RARβ在甲状腺乳头状癌中的表达与患者的年龄、性别、民族无明显相关关系（P〉0.05）。结论：DAPK1可能参与了甲状腺乳头状癌的发生发展过程。检测DAPK1和RARβ的表达水平可作为判断甲状腺乳头状癌转移和预后的参考指标。     

UbcH10 overexpression may represent a marker of anaplastic thyroid carcinomas

The hybridisation of an Affymetrix HG_U95Av2 oligonucleotide array with RNAs extracted from six human thyroid carcinoma cell lines and a normal human thyroid primary cell culture led us to the identification of the UbcH10 gene that was upregulated by 150-fold in all of the carcinoma cell lines in comparison to the primary culture cells of human normal thyroid origin. Immunohistochemical studies performed on paraffin-embedded tissue sections showed abundant UbcH10 levels in thyroid anaplastic carcinoma samples, whereas no detectable UbcH10 expression was observed in normal thyroid tissues, in adenomas and goiters. Papillary and follicular carcinomas were only weakly positive. These results were further confirmed by RT-PCR and Western blot analyses. The block of UbcH10 protein synthesis induced by RNA interference significantly reduced the growth rate of thyroid carcinoma cell lines. Taken together, these results would indicate that UbcH10 overexpression is involved in thyroid cell proliferation, and may represent a marker of thyroid anaplastic carcinomas.     

CD26 (dipeptidyl peptidase IV/DPP IV) as a novel molecular marker for differentiated thyroid carcinoma

In this report we show that CD26 (dipeptidyl peptidase IV/DPP IV) is a novel molecular marker for differentiated thyroid carcinoma. Northern-blot analysis of 22 various thyroid tissues revealed that CD26 is a more specific marker of differentiated thyroid carcinoma than 3 proto-oncogenes previously reported to increase mRNA expression in thyroid carcinomas: c-met, c-erbB-2 and EGF-R. A comparative study of 3 CD26 assays, Northern blotting, immunohistochemical staining and activity staining clearly showed that CD26 enzyme activity staining is the most specific assay for differentiated thyroid carcinoma, yet the easiest to perform. Activity staining of 216 thyroid tissues detected CD26 in all 52 papillary carcinomas and all 5 follicular carcinomas, while all 58 cases of Graves' disease were CD26-negative. Among benign neoplasms, 54 of 55 adenomatous goiters and 29 of 33 follicular adenomas were CD26-negative. Staining intensity of the enzyme activity was relative to the degree of CD26 mRNA expression. Southern-blot study showed no gene amplification or major translocation of the CD26 gene in 7 papillary carcinomas examined. Based on this study, ectopic expression of CD26 in differentiated thyroid carcinomas is thought to be mainly caused by increased CD26 mRNA expression. In conclusion, CD26 activity staining is a simple, specific assay which should be added to the usual pathological examinations in order to distinguish differentiated thyroid carcinomas from benign thyroid diseases. © 1995 Wiley-Liss, Inc.     

RET、HBME-1和Ki67在甲状腺良、恶性肿瘤中的表达

目的 探讨甲状腺良、恶性肿瘤中RET、HBME-1和Ki67基因蛋白的表达.方法 利用组织芯片技术采用二步法(PV-9000)进行免疫组化染色,检测22例结节性甲状腺肿、7例甲状腺滤泡性腺瘤,47例甲状腺乳头状癌、7例未分化癌、5例髓样癌、4例滤泡性癌、1例低分化岛状癌及1例鳞状细胞癌中上述3种蛋白的表达情况.结果 良性病变与恶性病变中3种蛋白的表达差异均有显著意义(P＜0.01),而良性病变中的不同类型间的差异无显著性(P＞0.05).HBME-1在甲状腺乳头状癌的阳性率高于滤泡性癌、髓样癌及未分化癌的表达,之间均有显著性差异(P＜0.01).Ki67在未分化癌阳性率高于乳头状癌的表达,二者比较差异显著(P＜0.05).结论 RET、HBME-1和Ki67的同时检测可作为良恶性甲状腺肿瘤鉴别诊断的指标,其中以HBME-1的敏感性最佳,RET次之,Ki67高表达可能是去分化、浸润和转移的标志.在甲状腺良性肿瘤中RET、HBME-1和Ki67同时阳性应视为恶变的高危状态.     

Critical role of cyclin D3 in TSH-dependent growth of thyrocytes and in hyperproliferative diseases of the thyroid gland

We report that cyclin D3 is rate limiting for G1 progression in thyroid follicular cells and that its constitutive upregulation by chronic stimulation of the TSH/cAMP pathway plays a role in human and experimental hyperproliferative diseases of the thyroid gland. These conclusions are supported by in vitro and in vivo studies. In rat thyrocytes (PC Cl 3 cells), cyclin D3 expression is enhanced in response to activation of the TSH/cAMP pathway. Interference with the expression of G1 cyclins (in particular cyclin D3) by the antisense methodology strongly reduced TSH-dependent proliferation of PC Cl 3 cells, indicating that proper progression through G1 requires cyclin D3. Accordingly, PC Cl 3 cells engineered to overexpress cyclin D3 (PC-D3 cells) show enhanced growth rate and elude hormone-dependence and contact inhibition. Using an animal experimental model of thyroid stimulation, we demonstrate that cyclin D3 is a key mediator of TSH-dependent proliferation of thyroid follicular cells also in vivo. Cyclin D3 protein levels were higher in the thyrocytes from glands of propylthiouracil-treated rats compared with control animals. The increase in cyclin D3 expression occurred after the propylthiouracil-induced increase in TSH levels and preceded the burst of cell proliferation. Finally, we found that cyclin D3 protein is expressed in a fraction of human goiters but it is strongly overexpressed in most follicular adenomas.     

Immunohistochemical detection of estrogen receptors in paraffin sections of human thyroid tissues.

The optimal demonstration of estrogen receptor binding in thyroid tissues was made under conditions of 10% protease in 50 mM Tris-HCl buffer (pH 7.6) for 10 min as the pretreatment digestion step, incubation of primary antibody (ER-ICA monoclonal kit; Abbott Laboratories) at 37 degrees C for 2 h and incubation of secondary antibody (ABC kit; Vector) at 37 degrees C for 40 min. Thyroid tissues used for assessing the reaction were 17 cases of goiter, 25 adenoma cases, 27 cases of papillary carcinoma, 14 cases of follicular carcinoma and 10 latent cancer cases. Incidences of positive estrogen receptor reaction were 22% (11/51) for all thyroid cancers, 20% (5/25) for the thyroid adenomas and 59% (10/17) for goiters. 15% (4/27) of papillary carcinomas, 21% (3/14) of follicular carcinomas and 40% (4/10) of latent cancers proved positive, the estrogen receptor reaction being limited to the nuclei of thyroid follicular/papillary type cells.     

Biological significance of aminopeptidase N/CD13 in thyroid carcinomas

Aminopeptidase N (APN)/CD13 is a transmembrane ectopeptidase expressed on a wide variety of cells. However, the precise function of APN/CD13 in tumor cells and the relationship of APN/CD13 to thyroid cancer remain unclear. In our study, we quantified the expression of APN/CD13 and additionally dipeptidyl peptidase IV (DPIV)/CD26 in thyroid carcinoma cell lines and in tissues of patients with thyroid carcinomas. Undifferentiated anaplastic thyroid carcinomas expressed more APN/CD13 than differentiated thyroid carcinomas. DPIV/CD26 showed an opposite expression pattern. We detected higher levels of DPIV/CD26 in follicular thyroid carcinomas (FTCs) and papillary thyroid carcinomas than in undifferentiated anaplastic thyroid carcinomas. In the undifferentiated thyroid carcinoma cell line 1736, APN/CD13 mRNA expression could be increased by epidermal growth factor, basic fibroblast growth factor, interleukin-6, and tumor necrosis factor alpha. FTC-133 cells stably transfected with an expression vector for APN-enhanced green fluorescent protein showed a higher migration rate than FTC-133 cells transfected with the enhanced green fluorescent protein-control plasmid. Overexpression of APN/CD13 in stably transfected cells is associated with down-regulation of N-myc down-regulated gene (NDRG)-1, melanoma-associated antigen ME491/CD63, and DPIV/CD26 gene expression. Inhibition of APN/CD13 mRNA expression by small interfering RNA induced NDRG-1, ME491/CD63, and DPIV/CD26 mRNA expression in cells of the undifferentiated thyroid carcinoma cell line C643. We conclude that APN/CD13-associated down-regulation of NDRG-1, ME491/CD63, and DPIV/CD26 in thyroid carcinoma cells is an important step of tumor progression to more malignant phenotypes, and we underline the important role of APN/CD13 as mediator in a multimolecular process regulating cell migration.     

Dissociation in the response of the adenylate cyclase system to thyrotropin and prostaglandin E2 in human thyroid carcinoma tissue

Because the existence of a damaged thyrotropin (TSH) receptor in thyroid tumors may be relevant in the perspective of a correct postsurgical therapy, the effect of TSH on cAMP intracellular accumulation in thyroid carcinoma (N = 16), follicular adenoma (N = 27) and normal tissue (N = 30) slices was studied and compared with that of nonspecific stimulus of thyroid adenylate cyclase-cAMP system, such as prostaglandin E2 (PGE2). While in all follicular adenomas a normal behavior of basal and post-TSH and -PGE2 stimulated cAMP accumulation was observed, basal cAMP levels were generally higher than in controls in 14 differentiated carcinomas, responses to TSH were reduced or absent, and response to PGE2 was close to normal. On the contrary, in two anaplastic carcinomas, both TSH and PGE2 produced a negligible modification of cAMP levels. Thus, in undifferentiated carcinomas, the adenylate cyclase-cAMP system seems to be altered; in differentiated carcinomas, the catalytic part of the system appears unaffected as it is PGE2-responsive. Therefore, some hypotheses are ruled out as an explanation for decreased sensitivity to TSH of differentiated carcinomatous cells.     

Thymosin beta-10 gene expression as a possible tool in diagnosis of thyroid neoplasias

Overexpression of thymosin beta-10 (TB10) has been shown in rat thyroid transformed cell lines, and in human thyroid carcinoma tissues and cell lines. To investigate whether TB10 detection could be a valid tool in the diagnosis of human thyroid neoplasias, we extended the analysis of TB10 expression to a large number of thyroid hyperproliferative and neoplastic tissues using an immunohistochemical assay. Our analyses showed a TB10 positive staining in all human thyroid carcinomas particularly in the anaplastic histotypes, whereas no TB10 immunostaining was observed in normal thyroid, in adenomas and the majority of the goiters. These results suggest that the evaluation of TB10 gene expression may be considered a promising means of diagnosis of human thyroid hyperproliferative disorders.     

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.