Expression of D2-40 in adjunct diagnosis of papillary thyroid carcinoma

The clinical pathologic criteria for nuclear features of papillary thyroid carcinoma are subjective and sometimes cannot distinguish carcinoma from adenomatous goiter and follicular neoplasms. No single antibody has demonstrated high sensitivity or specificity in making these distinctions. Using quantitative analysis of immunohistochemical staining with D2-40, a recently available monoclonal antibody used as a lymphatic endothelial marker, we examined 72 cases of papillary carcinoma. Controls included 36 follicular adenomas, 36 follicular carcinomas, and 20 adenomatous goiters with papillary hyperplasia. Cytoplasmic D2-40 immunoreactivity was present in 60 of 72 papillary carcinomas, 2 cases of follicular adenoma and 2 cases of follicular carcinoma, whereas no adenomatous goiter or normal thyroid glands contained positive epithelial cells. Overexpression of D2-40 in papillary thyroid carcinomas thus has potential diagnostic utility in differentiating these tumors from their potential histologic mimics.     

Insulin-like growth factor I in human thyroid tissue: Specific localization by immunohistochemistry andIn Situ hybridization

Insulin-like growth factor I and II (IGF-I and IGF-II) have been implicated in the replication of normal thyroid follicular cells in vitro. This study evaluates the distribution and abundance of immunoreactive IGF-I by histochemical analysis in human thyroid tissue with different histopathologic characteristics. We used two types of highly specific and sensitive polyclonal rabbit anti-IGF-I antibodies and one monoclonal antibody (MAb) with the immunoperoxidase technique on sections of 25 glands harboring adenomatous goiter; 11 glands with follicular adenoma (FA); 45 glands with thyroid carcinoma of papillary, follicular, and undifferentiated types; and 18 glands with Graves' disease. Immunoreactive IGF-I was present in some thyroid follicular cells of all thyroid tissues examined. The percentage of cells staining positively varies among the different processes, being lowest in normal thyroid tissues and highest in all thyroid carcinomas. The cytoplasmic pattern of IGF-I immunoreactivity also varied among the different thyroid conditions. Furthermore, using nonradioactivein situ hybridization (ISH) we detected IGF-I mRNA in the thyroid cells of adenomatous goiter. The expression was higher in the histologically hyperplastic areas. These findings provide further support for an autocrine and/or paracrine role of IGF-I in the function and/or growth of normal thyroid follicular cells and suggest that IGF-I may play a role in the dysfunctional growth of thyroid follicular cells in adenomatous goiter, thyroid carcinoma, and Graves' hyperthyroidism.     

The contributions of oestrogen receptor isoforms to the development of papillary and anaplastic thyroid carcinomas

Oestrogen (E2) is known to promote the proliferation of thyroid papillary carcinoma cells (KAT5). However, the molecular mechanism responsible is not well understood. In the study reported herein, the localization of ER alpha (ERalpha) and beta (ERbeta) in KAT5 and anaplastic carcinoma cells (FRO) was studied by immunofluorescence staining and by immunoblotting the proteins in subcellular fractions. Cell proliferation and apoptosis were also determined together with the expression of relevant proteins. The pattern of the subcellular localization of ERalpha and ERbeta differed between papillary and anaplastic cancer. Upon E2 treatment, the level of ERalpha increased in the nuclei of papillary cancer cells but ERbeta remained unchanged. The level of mitochondrial ERbeta surpassed that of ERalpha in anaplastic cancer cells. The different locations of ERalpha and ERbeta in KAT5 and FRO agreed with the finding that E2 promoted the proliferation of KAT5 but inhibited or did not affect that of FRO cells, and with the proposed functions of these two receptors. E2 inhibited the level of Bax in the mitochondria of papillary cancer, followed by a decrease of cytochrome c and/or apoptosis-inducing factor (AIF) release from the mitochondria into the cytosol. However, in anaplastic cancer, E2 promoted the expression of Bax in the mitochondria and the release of cytochrome c and/or AIF from mitochondria into the cytosol. Our results may explain the differences in epidemiology and responses to anti-tumour therapy between papillary and anaplastic cancer in terms of the subcellular localization of ER isoforms. In conclusion, the findings provide evidence to support the observation that E2 is an important factor in the development of thyroid cancer. The subcellular localization of ERalpha and ERbeta may account for the different pathogenesis of thyroid papillary and anaplastic cancers.     

c-Met expression of thyroid tissue with special reference to papillary carcinoma

It has become clear that papillary carclnomas of the thyroid often express the receptor for c-Met/hepatocyte growth factor (HGF) receptor, but little Is known about the role of the HGF and c-Met system In the pathogenesis of thyroid carcinoma. In this study, the expression of c-MeUHGF receptor was evaluated in thyroid tlssue by western blot and immunohistochemistry, and compared with the concentration of HGF. Clinicopathologlcal characteristics were also compared. Fmeen of 20 papillary Carcinomas (75%) showed eMet bands of 145 kDa. No or only a low frequency of c-Met expression was detected in healthy thyroid tissue (0/5), thyroiditis or Basedow's disease (0/2), adenomatous goiters (0/8), follicular adenomas (119, 11%) and undifferentiated carcinomas (0/2). These results were confirmed by immunohistochemistry, but a relatively higher frequency of c-Met expression was detected in adenomatous goiters (25%), follicular adenoma (44%) ancl papillary Carcinoma (100%) using formalin-fixed and paraffin-embedded materials. A strong immunoreaction for c-Met was observed In the tumor cytoplasm of paplllary carcinomas among the fibrous tissues situated at the perlphery of the tumor. The densito-metrically measured expression of c-Met had no relation to tumor stage in papillary carcinoma, but did correlate to the concentration of HGF in papillary carcinomas. In conclusion, in thyroid lesions, c-Met was highly expressed specifically in the cytoplasm of papillary carcinomas. c-Met expression was not related to the aggressiveness of the tumor but was related to the concentration of HGF, which was probably derived from the stroma. Also, the c-Met system might play a role in the pathogenesis of papillary carcinoma of the thyroid.     

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。     

Expression of intermediate filament proteins in normal and diseased thyroid glands.

A total of 67 samples from normal and pathological thyroid glands were stained (as formalin fixed paraffin sections) with a panel of monoclonal antibodies directed against intermediate filament proteins. The study confirmed previous reports of cytokeratin and vimentin coexpression in primary thyroid carcinomas, but coexpression was also detected in normal thyroid and in a range of benign conditions including follicular adenomas, Hashimoto's thyroiditis, and diffuse hyperplasia (thyrotoxicosis). Prekeratin expression was found (using antibodies recognising higher molecular weight cytokeratins) predominantly in areas of squamous change, independent of the underlying thyroid pathology. This study does not therefore support previous findings that prekeratin expression provides a reliable means of distinguishing follicular pattern papillary carcinoma from follicular carcinoma with its poorer prognosis or that it helps distinguish benign from malignant papillary lesions. No evidence of desmin or neurofilament expression was seen, and in particular, neurofilaments could not be detected in any of the cases of medullary carcinoma studied.     

Expression of intermediate filament proteins in thyroid gland and thyroid tumors

The presence of intermediate filament proteins of cytokeratin/prekeratin type and vimentin type was evaluated in non-neoplastic thyroid glands and in different types of thyroid neoplasms. Follicular epithelium of both normal and goitrous thyroids showed a strong reaction with anticytokeratin antibodies that widely cross-react with various simple epithelia. On the other hand, in normal thyroid, there were only occasionally (in one of 12 cases) solitary cells reacting with antibodies to epidermal prekeratin. In nodular goiters, such cells were often seen (eight of 18), especially among the lining cells of cysts, and in chronic thyroiditis in all (12 of 12) cases. Only the stromal cells and intraluminal macrophages reacted with antibodies to vimentin. Neoplastic cells of papillary carcinomas showed a positive staining reaction both with antibodies to cytokeratins and to epidermal prekeratin. Follicular carcinoma cells, although positive for cytokeratins, could generally not be stained with antibodies to epidermal prekeratin. Medullary carcinoma cells also showed cytokeratin positivity and, only occasionally, positivity for epidermal prekeratin. Anaplastic carcinomas were also reactive with antibodies to cytokeratin but, for the most part, were negative for epidermal prekeratin. Interestingly, some neoplastic cells of all types of thyroid carcinomas also appeared to contain vimentin, as shown with both polyclonal and monoclonal antivimentin antibodies. In contrast to carcinomas, the intermediate filaments of thyroid sarcomas and lymphomas were only of vimentin type. Furthermore, it was found that the papillary structures in benign goiters were only reactive with cytokeratin antibodies and lacked, in contrast to papillary carcinomas, epidermal prekeratin-like immunoreactivity. Hence, the analysis of intermediate filament proteins of thyroid tumors can be utilized to differentiate between papillary and follicular carcinomas and between benign and malignant papillary lesions as well as between anaplastic thyroid carcinomas and sarcomas or lymphomas.     

NDRG1 protein overexpression in malignant thyroid neoplasms

OBJECTIVES:

The aim of this study was to examine the expression of the N-myc downstream-regulated gene 1 protein in benign and malignant lesions of the thyroid gland by immunohistochemistry.

INTRODUCTION:

N-myc downstream-regulated gene 1 encodes a protein whose expression is induced by various stimuli, including cell differentiation, exposure to heavy metals, hypoxia, and DNA damage. Increased N-myc downstream-regulated gene 1 expression has been detected in various types of tumors, but the role of N-myc downstream-regulated gene 1 expression in thyroid lesions remains to be determined.

METHODS:

A tissue microarray paraffin block containing 265 tissue fragments corresponding to normal thyroid, nodular goiter, follicular adenoma, papillary thyroid carcinoma (classical pattern and follicular variant), follicular carcinoma, and metastases of papillary and follicular thyroid carcinomas were analyzed by immunohistochemistry using a polyclonal anti- N-myc downstream-regulated gene 1 antibody.

RESULTS:

The immunohistochemical expression of N-myc downstream-regulated gene 1 was higher in carcinomas compared to normal thyroid glands and nodular goiters, with higher expression in classical papillary thyroid carcinomas and metastases of thyroid carcinomas (P < 0.001). A combined analysis showed higher immunohistochemical expression of NDRG1 in malignant lesions (classical pattern and follicular variant of papillary thyroid carcinomas, follicular carcinomas, and metastases of thyroid carcinomas) compared to benign thyroid lesions (goiter and follicular adenomas) (P  =  0.043). In thyroid carcinomas, N-myc downstream-regulated gene 1 expression was significantly correlated with a more advanced TNM stage (P  =  0.007) and age, metastasis, tumor extent, and size (AMES) high-risk group (P  =  0.012).

CONCLUSIONS:

Thyroid carcinomas showed increased immunohistochemical N-myc downstream-regulated gene 1 expression compared to normal and benign thyroid lesions and is correlated with more advanced tumor stages.     

Distribution of sialic acid-dependent carbohydrate epitope in thyroid tumors: Immunoreactivity of FB21 in paraffin-embedded tissue sections

The reactivity of monoclonal antibody FB21, which recognizes a sialic acid-dependent carbohydrate epitope, was tested with 94 non-neoplastic and neoplastic thyroid tissue specimens using immunohistochemical methods on formalin-fixed, paraffin-embedded tissue sections. These specimens included 11 cases of adenomatous goiter, three cases of Basedow's disease, 30 cases of follicular adenoma, 20 cases of papillary carcinoma, 15 cases of follicular carcinoma, six cases of medullary carcinoma, and nine cases of anaplastic carcinoma. FB21 reacted with 14 of 15 cases of follicular carcinoma that showed a microfollicular or trabecular pattern, and with nine of 20 cases of papillary carcinoma. A positive reaction was found on the cell surface membranes or apical parts of neoplastic follicles. FB21 also reacted with five of 30 cases of follicular adenoma. These cases showed a follicular pattern and positive staining pattern similar to that in follicular carcinoma. Adenomatous goiters, Basedow's disease, medullary carcinomas, and anaplastic carcinomas were negative for FB21 reactivity. Although the different reactivities of FB21 with papillary carcinoma and follicular carcinoma remain to be investigated, the high frequency of reactivity with FB21 suggests that it may be useful as a complement to morphological diagnosis in follicular carcinoma.     

Diagnostic utility of galectin-3 and CD26/DPPIV as preoperative diagnostic markers for thyroid nodules

The aim of this study was to search for diagnostic markers that could correctly identify thyroid nodular lesions requiring urgent surgical treatment. We investigated whether galectin-3 and dipeptidyl peptidase IV (CD26/DPPIV) could be potential markers for improving the diagnostic accuracy of conventional cytology. Seventy-nine patients with histologically proven thyroid diseases were analyzed. The immunocytochemical staining results showed galectin-3 expression in neoplastic cells of all 37 papillary carcinomas, five of six follicular carcinomas, all three anaplastic carcinomas, one of three medullary carcinomas, and two of 14 follicular adenomas. All 16 adenomatous goiters were negative for galectin-3 immunostaining. On the other hand, all 37 papillary carcinomas, all six follicular carcinomas, and one of three anaplastic carcinomas revealed CD26/DPPIV expression, whereas all three medullary carcinomas were negative. Among benign thyroid lesions, four of 14 follicular adenomas and two of 16 adenomatous goiters exhibited varying degrees of immunoreactivity for CD26/DPPIV. RT-PCR analysis demonstrated overexpression of galectin-3 and CD26/DPPIV mRNAs in all six papillary and all three follicular carcinomas analyzed, whereas the mRNA expressions of these molecules were barely or not detectable in benign thyroid lesions and normal thyroid tissues, except for one case of follicular adenoma. In conclusion, we demonstrate that galectin-3 and CD26/DPPIV were consistently coexpressed at protein and mRNA levels in differentiated thyroid carcinomas. We propose that combined immunostaining for galectin-3 and CD26/DPPIV in the preoperative evaluation of thyroid nodules may play a role in accurate cytodiagnosis.     

Presence of immunoreactive corticotropin releasing hormone in thyroid lesions.

Corticotropin-releasing hormone (CRH) functions as a regulator of the hypothalamic-pituitary-adrenal axis and coordinator of the stress response. Immunoreactive CRH (IrCRH) is also produced in a variety of inflammatory sites, where this peptide acts as a proinflammatory cytokine. To detect CRH in autoimmune thyroid disease as well as in disorders that may be associated with an inflammatory reaction within this gland, we examined immunohistochemically 45 thyroid lesions, including 12 nodular goiters, 9 cases of Hashimoto thyroiditis, 6 follicular adenomas, 4 follicular and 8 papillary carcinomas, 4 Hürthle cell tumors, 1 medullary cancer, and 1 insular thyroid carcinoma. We also examined the presence of IrCRH in the adjacent normal thyroid parenchyma. The avidin-biotin complex method was employed on formalin-fixed, paraffin-embedded tissue, using a highly specific, affinity-purified polyclonal rabbit anti-CRH antibody. Granular cytoplasmic immunostaining of follicular cells was observed in 100% of the cases of Hashimoto thyroiditis, 77% of the neoplasms and 42% of goiters. The intensity of the staining was more pronounced in Hashimoto thyroiditis and Hürthle cell tumors, whereas the remaining lesions exhibited a heterogeneous staining pattern. No IrCRH was observed in the normal thyroid parenchyma. Using a specific radioimmunoassay, the IrCRH in extracts of simple thyroid goiters, papillary carcinomas, and Hürthle cell tumors ranged between 0.031 and 0.224 pmol/g of wet tissue but was undetectable in normal thyroid parenchyma. The IrCRH molecule in the thyroid gland eluted at the same fraction as synthetic rat/human CRH 1-41 in reverse phase high pressure liquid chromatography. We conclude that IrCRH is present in thyroid lesions, predominantly in those related to autoimmune phenomena, suggesting that this neuropeptide may be directly and/or indirectly involved with inflammatory processes taking place in this gland.     

Ultrastructural observations on the capillaries of human thyroid tumours.

Ultrastructure of the capillaries of malignant and benign thyroid tumours has been examined. The material consisted of biopsies from six cases of thyroid papillary carcinoma, one case of follicular (foetal type) adenoma and six cases of nodular adenomatous goitre. In the group of nodular adenomatous goitre and in the follicular adenoma, the capillary wall was made up of fenestrated endothelium similar to that of capillaries of normal human thyroid. The fenestrae occupied a large area of the endothelial wall. Micro- and macropinocytotic vesicles were frequent in the endothelial cytoplasm. In the thyroid carcinomas the papillary structures always contained numerous capillaries with fenestrated endothelium. The microfollicular area and the solid tumoral areas of the papillary carcinoma showed occasional capillaries with fenestrated endothelium, but many capillaries were lined with continuous endothelium. The capillaries in all the specimens were surrounded externally by a continuous basement membrane which was frequently bilaminate or multilaminate. This study indicates that capillaries with fenestrated endothelium are characteristic of thyroid tumours which arise from follicular cells.     

Lectin histochemistry of papillary and follicular carcinoma of the thyroid gland

The lectin-binding properties of human follicular and papillary carcinoma were studied histochemically and compared with lectin binding to normal or goitrous thyroid tissue. Well-differentiated minimally invasive follicular carcinoma showed a lectin-binding pattern essentially identical to those of the normal thyroid gland and benign adenomatous lesions. Overtly invasive follicular carcinoma showed focal reactivity with some lectins that were nonreactive with normal follicular thyroid cells (Solanum tuberosum and soybean in three of three cases; Ulex europaeus in two of three cases; and Dolichos biflorus, Laburnum alpinum, and peanut in one of three cases). In papillary carcinomas, the cells lining the papillary structures reacted focally with some lectins that did not bind to normal thyroid cells (S tuberosum and U europeaus in seven of seven cases; Helix pomatia, Helix aspersa, and soybean in four of seven cases; and peanut, Griffonia simplicifolia, D biflorus, and Vicia villosa in one of seven cases). All these lectins, as well as those reacting with normal thyroid cells, reacted more strongly with cells of papillary structures than with those forming solid nests and follicles. Despite these lectin-defined differences in the composition of glycoconjugates of benign and malignant thyroid cells, the inconsistent and focal nature of the changes precludes the use of lectins in diagnostic histopathology.     

Quantitative nuclear cell image analyses of thyroid tumors from archival material

Sixty-three sections of Feulgen-stained thyroid cell nuclei from paraffin-embedded material, including five multinodular goiters, 10 adenomas, 36 papillary carcinomas, seven follicular carcinomas, and five medullary carcinomas were analyzed by means of the SAMBA 200 (TITN, Grenoble, France) cell image processor. This was done in order to obtain nuclear characteristics of papillary versus follicular carcinomas. The nuclear features were assessed by morphometric, densitometric, and textural parameters. Our preliminary results indicate that the cell nuclei from typical histopathologic specimens of follicular thyroid cancers belong to a larger thyroid cell nuclei population corresponding to the histopathologic family of papillary thyroid cancers. This follicular neoplastic cell nuclei population appears to be quite distinct from the typical medullary neoplastic cell nuclei population which also belongs to the papillary neoplastic cell nuclei population. It appears that there is a specific papillary cell nuclei subpopulation containing typical hypochromatic cell nuclei. We also observed a dramatic increase in nuclear size and hyperchromatism between normal (multinodular goiters) and neoplastic (carcinomas) thyroid tissues, with the benign tissues (adenomas) showing intermediate nuclear characteristics.     

Nuclear grooves: A useful criterion in the cytopathologic diagnosis of papillary thyroid carcinoma

A recent report emphasized the usefulness of the grooved nucleus as a diagnostic criterion of papillary thyroid carcinoma (PTC) in histopathologic material. The present study was undertaken to evaluate whether grooved nuclei can serve as an additional diagnostic criterion for PTC in cytologic material obtained by fine-needle aspiration (FNA). Slides from 124 consecutive thyroid FNAs were reviewed. Specimens included 11 PTCs, one follicular carcinoma, six follicular adenomas, eight follicular neoplasms not otherwise specified, 10 cases of chronic thyroiditis, and 88 colloid nodules/adenomatous goiters. Among the PTC cases, grooved nuclei were found in all 11 (100%), intranuclear inclusions in nine (82%), papillary fragments in seven (64%), and psammoma bodies in two (18%). Nuclear grooves were also observed in two of the 113 non-PTC cases (1.8%), both of which were colloid nodules, one with extensive Hurthle-cell change. The grooved nuclei were best identified on Papanicolaou-stained material. They were inconspicuous and difficult to identify in air-dried Diff-Quik-stained material. It appears that the recognition of grooved nuclei among tumor cells is a valuable diagnostic feature of PTC in cytologic material stained with polychromatic Papani-colaou stain.     

甲状腺肿瘤3号染色体短臂杂合性缺失的研究

目的 探讨甲状腺肿瘤中3号染色体短臂(3p)杂合性缺失(LOH)状态及其临床意义.方法 收集74例甲状腺肿瘤标本,包括20例甲状腺腺瘤(FA)、24例滤泡性甲状腺癌(FTC)和30例乳头状甲状腺癌(Prc).通过PCR扩增和银染分析其3p上11个微卫星位点的杂合性缺失状态.结果 FFC的LOH频率达到71%(17/24),PTC中30%(9/30),FA中10%(2/20).FFC的3p LOH频率显著高于FA和PTC(P<0.01).FTC中存在两个最小共同缺失区,分别位于3p26-pter和3p14.2-3p22.PTC上存在一个最小共同缺失区,位于3p 25.2-26.1.结论 FTC的3p LOH频率显著高于FA和PTC.3p的3个最小缺失区上可能存在着与FTC和PTC发生发展相关的肿瘤抑制基因.