Immunohistochemical analysis of survivin expression in thyroid follicular adenoma and carcinoma.

Survivin is one of the 8 members of human inhibitor of apoptosis , which is differentially expressed in cancerous/transformed cells versus normal differentiated tissues. This retrospective study of thyroid histologic samples aimed to assess the clinical usefulness of survivin immunostaining for discrimination between follicular adenoma and carcinoma of thyroid. Immunohistochemical staining for survivin was performed on 41 lesions from patients who had undergone surgery for either follicular adenoma or carcinoma of thyroid. Survivin expression was significantly (P < 0.005) higher in the cases that received a diagnosis of carcinoma in comparison with follicular adenomas cases. Odds ratio of follicular carcinoma for survivin expression was 21.375 (95% CI: 3.283 to 139.177). Our results showed potential value of survivin in discrimination between follicular thyroid adenoma and follicular thyroid carcinoma. We conclude that survivin is a potential candidate for further investigation in the proper histologic diagnosis of thyroid cancers.     

Differential expression of galectin-1 and galectin-3 in thyroid tumors. Potential diagnostic implications.

Carcinoma of the thyroid gland, the most frequently diagnosed endocrine malignancy, is often associated with early regional metastases. With the exception of papillary carcinoma, distinguishing benign from malignant thyroid neoplasms in the absence of metastatic disease is difficult. Recently, the vertebrate lectins galectin-1 and galectin-3 have been implicated in the regulation of cellular growth, differentiation, and malignant transformation of a variety of tissues. To determine whether these galectins have a role in thyroid neoplasia, we analyzed 32 specimens from thyroid malignancies (16 papillary, 7 follicular, 8 medullary carcinomas, and 1 metastasis to lymph node), 10 benign thyroid adenomas, 1 nodular goiter, and 33 specimens from adjacent normal thyroid tissue for the expression of galectin-1 and galectin-3 with immunohistochemical and immunoblotting techniques utilizing anti-galectin antibodies. All thyroid malignancies of epithelial origin (ie, papillary and follicular carcinomas) and a metastatic lymph node from a papillary carcinoma expressed high levels of both galectin-1 and galectin-3. The medullary thyroid carcinomas, which are of parafollicular C cell origin, showed a weaker and variable expression of these galectins. In contrast, neither benign thyroid adenomas nor adjacent normal thyroid tissue expressed galectin-1 or galectin-3. These results suggest that galectin-1 and galectin-3 may be associated with malignant transformation of thyroid epithelium and may potentially serve as markers for distinguishing benign thyroid adenomas from differentiated thyroid carcinomas.     

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.     

Expression of survivin is associated with malignant potential in epithelial ovarian carcinoma

Survivin is a new member of the inhibitor of apoptosis family of anti-apoptotic proteins. It has been reported that survivin is expressed during fetal development and in cancer tissues. Because suppression of apoptosis is important for carcinogenesis and tumor growth, we investigated the expression of survivin in human endometrial carcinomas. We analyzed serial frozen sections for survivin protein expression in 26 patients with ovarian epithelial carcinoma and 10 patients with benign cystadenoma of the ovary by fluorescent immunohistochemistry. We analyzed the relationship between the percentages of survivin-stained cells and the characteristics of the patient including histological classification, clinical stage, histological grade, and clinical outcome. Survivin was weakly detected in some benign ovarian cystadenomas (0-12.1%). There was, however, abundant survivin immunoreactivity in the nucleus and/or cytoplasm of the epithelial ovarian carcinoma cells. Scoring on the basis of the percentage of positive cells indicated that survivin expression was significantly associated with PCNA-labeling index, clinical stage, histological grade, clinical outcome, and survival rate (p<0.01, respectively). We conclude that the survivin protein is a defining diagnostic marker for epithelial ovarian carcinomas that may also yield prognostic information.     

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.     

S100A4蛋白在甲状腺癌中的表达和意义

 目的 探讨S100A4蛋白在甲状腺癌组织中的表达及临床意义。方法 应用免疫组织化学SP法,检测56例甲状腺癌（乳头状腺癌30例,髓样癌12例,滤泡状癌8例,未分化癌6例）、45例甲状腺良性病变（甲状腺腺瘤、结节性甲状腺肿、原发性甲状腺功能亢进各15例）及13例甲状腺癌癌旁正常组织中S100A4蛋白的表达水平。结果 甲状腺癌组织中S100A4蛋白表达阳性率为83.9%（47/56）,表达强度呈（+）者25例,（++）者12例,（+++）者10例;甲状腺良性病变组织中S100A4蛋白表达阳性率为11.1%（5/45）,表达强度均为（+）;癌旁正常甲状腺组织中S100A4蛋白表达均呈阴性。甲状腺癌组织与后两者比较,差异均具有统计学意义（P<0.01）。临床分期Ⅰ～Ⅱ期者S100A4蛋白表达阳性率为77.8%（28/36）,表达强度为（+）者22例,（++）者4例,（+++）者2例;临床分期Ⅲ～Ⅴ期者表达阳性率为95.0%（19/20）,表达强度为（+）者3例、（++）者8例、（+++）者8例;两者间差异有统计学意义（P<0.01）。有淋巴结转移者S100A4蛋白表达阳性率为100%（16/16）,表达强度为（++）者8例、（+++）者8例;无淋巴结转移者表达阳性率为77.5%（31/40）,表达强度为（+）者25例、（++）者4例、（+++）者2例;两者间差异有统计学意义（P<0.01）。不同病理类型甲状腺癌组织中S100A4蛋白表达阳性率和表达强度差异无统计学意义（P>0.05）。结论 S100A4蛋白表达与甲状腺肿瘤细胞的恶性增殖和侵袭转移密切相关。S100A4蛋白表达强度作为判断甲状腺癌的恶性程度及预后的检测指标,具有临床应用的价值。     

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。     

RNA interference targeting CD147 inhibits the proliferation,invasiveness, and metastatic activity of thyroid carcinoma cells by down-regulating glycolysis

A high rate of glycolytic flux, even in the presence of oxygen, is a key metabolic hallmark of cancer cells. Lactate, the end product of glycolysis, decreases the extracellular pH and contributes to the proliferation, invasiveness and metastasis of tumor cells. CD147 play a crucial role in tumorigenicity, invasion and metastasis; and CD147 also interacts strongly and specifically with monocarboxylate transporter1 (MCT1) that mediates the transport of lactate. The objective of this study was to determine whether CD147 is involved, via its association with MCT1 to transport lactate, in glycolysis, contributing to the progression of thyroid carcinoma. The expression levels of CD147 in surgical specimens of normal thyroid, nodular goiter (NG), well-differentiated thyroid carcinoma (WDTC), and undifferentiated thyroid carcinoma (UDTC) were determined using immunohistochemical techniques. The effects of CD147 silencing on cell proliferation, invasiveness, metastasis, co-localization with MCT1, glycolysis rate and extracellular pH of thyroid cancer cells (WRO and FRO cell lines) were measured after CD147 was knocked-down using siRNA targeting CD147. Immunohistochemical analysis of thyroid carcinoma (TC) tissues revealed significant increases in signal for CD147 compared with normal tissue or NG, while UDTC expressed remarkably higher levels of CD147 compared with WDTC. Furthermore, silencing of CD147 in TC cells clearly abrogated the expression of MCT1 and its co-localization with CD147 and dramatically decreased both the glycolysis rate and extracellular pH. Thus, cell proliferation, invasiveness, and metastasis were all significantly decreased by siRNA. These results demonstrate in vitro that the expression of CD147 correlates with the degree of dedifferentiation of thyroid cancer, and show that CD147 interacts with MCT1 to regulate tumor cell glycolysis, resulting in the progression of thyroid carcinoma.     

Prognostic significance of anti-apoptosis proteins survivin and bcl-2 in non-small cell lung carcinomas: a clinicopathologic study of 102 cases.

Inhibitors of apoptosis, including bcl-2 and survivin (a novel gene encoding a unique apoptosis inhibitor), regulate cell proliferation by promoting cell survival. Although survivin has been detected in several human cancers, its prognostic significance and relationship to bcl-2 are not well characterized in lung cancer. Tissue sections from 102 non-small cell lung carcinomas (NSCLC) were immunostained using antibodies against survivin and bcl-2. Staining results were correlated with prognostic variables. Immunoreactivity for survivin and bcl-2 was observed in 53% and 21% of NSCLCs, respectively. Fifty-two percent of the 50 squamous cell carcinomas and 54% of the 52 adenocarcinomas expressed survivin. Survivin positivity correlated with tumor stage in squamous cell carcinoma. On univariate analysis, survivin expression correlated with decreased patient survival in NSCLC and in the subset of squamous cell carcinomas, but not in adenocarcinomas. On multivariate analysis, survivin was an independent predictor, along with distant metastasis and large tumor size. Eighteen percent of squamous cell carcinomas and 24% of adenocarcinomas expressed bcl-2. On univariate analysis, bcl-2 expression correlated with increased patient survival in NSCLC and in the subset of squamous cell carcinomas. An inverse correlation between the expression of survivin and bcl-2 was noted. Survivin immunoreactivity is an independent predictor of shortened survival in NSCLC, while bcl-2 protein expression correlated with prolonged patient survival. These findings indicate an inverse relationship between survivin and bcl-2 expression and suggest that these two inhibitors of apoptosis function through different pathways in the regulation of tumorigenesis in NSCLC.     

Frequency of medullary thyroid carcinoma in an endemic goiter area

A series of 41 poorly differentiated follicular carcinomas of the thyroid gland without histopathological features of medullary or papillary carcinoma and 9 cases of undifferentiated thyroid carcinoma of the small cell type (diagnosed between 1967 and 1983) were investigated immunohistochemically. Two poorly differentiated follicular carcinomas showed a considerable number of calcitonin-positive cells in addition to the weakly thyroglobulin-positive tumor cells. One of these cases revealed several areas with calcitonin-positive tumor cells with additional squamous metaplasia with keratinization. No medullary carcinomas could be demonstrated among the 9 cases previously diagnosed as undifferentiated thyroid carcinomas of the small cell type. From the epidemiological point of view the application of immunohistochemistry does not significantly increase the proportion of medullary carcinomas detected in our endemic goiter area. The incidence of medullary carcinoma remains surprisingly low when compared with nonendemic areas.     

Immunohistochemical Demonstration of Epidermal Growth Factor Receptor and Ceruloplasmin in Thyroid Diseases

The expression of epidermal growth factor receptors (EGFR) and ceruloplasmin (CP) in thyroid diseases was investigated by immunohistochemical methods, and the results were compared with the expression of thyroglobulin (TG). Eighty eight surgical specimens of thyroid diseases, including follicular carcinoma (7 cases), papillary carcinoma (20 cases), follicular adenoma (29 cases), adenomatous goiter (10 cases), diffuse hyperplasia (20 cases) and chronic thyroiditis (2 cases), were studied. All cases of follicular carcinoma and 18 cases (90%) of papillary carcinoma expressed the EGFR immunoreaction in the cytoplasm with a moderate to strong staining intensity. A weak immunoreaction for EGFR was noted in some benign thyroid diseases. CP showed various degrees of positivity in all cases of follicular carcinoma and 19 cases (95%) of papillary carcinoma. The benign thyroid lesions were consistently negative for this antigen, not counting one case of Hiirthle cell adenoma. There was a positive correlation between EGFR and CP immunostaining intensity in thyroid carcinomas, representing higher expression of EGFR accompanied by a stronger staining intensity of CP. Except for two cases of papillary carcinoma, all cases showed immunoreaction for TG. The results indicate the enhanced expression of EGFR and CP in thyroid carcinomas. EGFR and CP thus appear to be valuable tools for differential diagnosis between benign and malignant thyroid neoplasms.     

Immunohistochemical demonstration of epidermal growth factor receptor and ceruloplasmin in thyroid diseases

The expression of epidermal growth factor receptors (EGFR) and ceruloplasmin (CP) in thyroid diseases was investigated by immunohistochemical methods, and the results were compared with the expression of thyroglobulin (TG). Eighty-eight surgical specimens of thyroid diseases, including follicular carcinoma (7 cases), papillary carcinoma (20 cases), follicular adenoma (29 cases), adenomatous goiter (10 cases), diffuse hyperplasia (20 cases) and chronic thyroiditis (2 cases), were studied. All cases of follicular carcinoma and 18 cases (90%) of papillary carcinoma expressed the EGFR immunoreaction in the cytoplasm with a moderate to strong staining intensity. A weak immunoreaction for EGFR was noted in some benign thyroid diseases. CP showed various degrees of positivity in all cases of follicular carcinoma and 19 cases (95%) of papillary carcinoma. The benign thyroid lesions were consistently negative for this antigen, not counting one case of Hürthle cell adenoma. There was a positive correlation between EGFR and CP immunostaining intensity in thyroid carcinomas, representing higher expression of EGFR accompanied by a stronger staining intensity of CP. Except for two cases of papillary carcinoma, all cases showed immunoreaction for TG. The results indicate the enhanced expression of EGFR and CP in thyroid carcinomas. EGFR and CP thus appear to be valuable tools for differential diagnosis between benign and malignant thyroid neoplasms.     

Expression of sodium iodide symporter in benign and malignant human thyroid tissues

The extent of human sodium iodide symporter (hNIS) expression in different kinds of human thyroid cancer tissues and cell lines remains controversial. In this study, polyclonal antibodies to hNIS were used to analyze the expression of symporter protein in benign and malignant human thyroid tissues. Formalin-fixed, paraffin wax-embedded tissue sections were used. Staining was performed using primary polyclonal antibody of rabbit anti-human hNIS diluted in PBS (1:500). Results showed that 2 of 3 normal tissue, 3 of nodular hyperplasia, one follicular adenoma, 3 of 11 papillary thyroid carcinoma, 1 of 5 follicular carcinoma and none of 3 metastatic thyroid epithelial tissue specimens stained positively for hNIS. A higher percentage of positive staining for symporter protein was found in benign thyroid tissues including normal thyroid tissue, nodular hyperplasia, and adenoma (60%). In contrast, papillary and follicular thyroid carcinomas demonstrated lower symporter protein expression (20%). In conclusion, although the number of tissue samples examined in this study was small, hNIS staining found a higher ratio of symporter protein expression in normal and benign thyroid tissues compared with malignant tissues. Determination of the reason for discrepancies in the expression of hNIS in in vivo and in vitro studies will require further investigation.     

Galectin-3 does not reliably distinguish benign from malignant thyroid neoplasms

AIMS: To determine whether galectin-3 is a sensitive indicator of thyroid malignancy. It has been suggested as a potential marker for differentiating thyroid carcinoma from benign or non-neoplastic lesions in preoperative fine-needle aspirates (FNAs). METHODS: Galectin-3 protein expression was assessed by immunohistochemistry in formalin-fixed thyroid tissues from 124 patients with histological diagnoses of papillary carcinoma (n = 38), follicular carcinoma (n = 19), follicular adenoma (n = 32) and dominant nodules of multinodular goitre (n = 35). Expression of galectin-3 was also assessed by Western blotting in 24 fresh thyroid tissues. RESULTS: Galectin-3 expression was observed in the majority of carcinomas (papillary 92%; follicular 74%). However, a large proportion of follicular adenomas (72%) and multinodular goitres (57%) also expressed galectin-3. In addition, galectin-3 expression was observed in epithelial cells of normal thyroid tissue and Hashimoto's thyroiditis. Galectin-3 immunopositivity was significantly greater in papillary carcinomas than in dominant nodules or follicular adenomas (P < 0.0001, P = 0.0005, respectively). However, galectin-3 expression was no greater in follicular carcinomas than in follicular adenomas (P = 0.8735). Western blotting analysis confirmed both the specificity of the antiserum and expression of galectin-3 in multinodular goitres, follicular adenomas/carcinomas and papillary carcinomas. CONCLUSION: The data demonstrate that galectin-3 is not a reliable immunohistochemical marker to distinguish benign from malignant thyroid follicular lesions.     

Enhanced B-Raf protein expression is independent of V600E mutant status in thyroid carcinomas

BRAF (7q24) encodes a serine/threonine protein kinase, and its expression level varies in different tissues. Although a high prevalence of BRAF mutation has been suggested as an important event in thyroid tumorigenesis, little is known about the expression pattern of B-Raf in the thyroid. Thus, we examined the expression of B-Raf in various neoplastic and nonneoplastic thyroid tissues and compared it with BRAF mutational status. Normal and hyperplastic thyroid tissues showed focal and faint immunoreactivity for B-Raf, especially in cuboidal follicular cells of small follicles. In contrast, diffuse expression of B-Raf was observed in follicular adenomas and well-differentiated carcinomas. The missense point mutation BRAF(V600E) was identified in 42% (13/31 cases) of papillary carcinomas and 33% (5/15 cases) of undifferentiated carcinomas but not in normal thyroid tissues, nodular hyperplasia, follicular adenomas, or follicular carcinomas. The immunohistochemical expression of B-Raf did not correlate with BRAF mutational status. Moreover, Western blotting revealed that B-Raf expression in thyroid carcinoma cell lines was also independent of BRAF mutation. Serum or fibroblast growth factor-1 stimulation further activates ERK1/2 in heterozygous BRAF(V600E)-positive carcinoma cells as well as BRAF(V600E) mutation-negative carcinoma cells. In conclusion, heterogeneous focal expression of wild-type B-Raf in nonneoplastic tissues may play a role in the growth or functional activity of thyroid follicular cells. In contrast, diffuse expression of wild-type and/or mutant B-Raf may be involved in the tumorigenic process resulting in activation of the mitogen-activated protein kinase signaling pathway in cooperation with other genetic abnormalities and activation of ligand-receptor signaling pathways.     

Differential gene expression profiling of aggressive and nonaggressive follicular carcinomas

The classification of follicular thyroid neoplasms requires surgical resection for histologic evaluation of malignancy. Because variable clinical behavior exists, genomic expression profiling may lead to the identification of novel markers that facilitate better biologic classification. We performed for the first time gene expression analysis on clinically aggressive and nonaggressive follicular carcinomas (FCs) from patients for whom long-term follow-up data were available. We examined matched fresh-frozen tissue from 15 histopathologically diagnosed follicular carcinomas (7 patients with documented distant metastasis and/or death from disease and 8 patients without recurrence). For categorical comparison, we analyzed 4 follicular adenomas (FAs). The biologic control comprised 11 normal thyroid tissue specimens. High-quality RNA was extracted from the tissues, labeled, and hybridized to an Affymetrix (Santa Clara, CA) oligonucleotide microarray (HG-U133A). With the exceptions of 1 follicular adenoma and 1 follicular carcinoma, unsupervised hierarchical cluster analysis revealed 2 distinct groups--one containing normal thyroid tissue and follicular adenomas and another containing follicular carcinomas. We identified 421 genes that were differentially expressed between histologically normal thyroid tissues and all follicular neoplasms (P < 0.01; fold-change >2), 94 genes that distinguished follicular carcinomas from follicular adenomas (including PBP and CKS2), and 4 genes that distinguished aggressive follicular carcinomas from nonaggressive follicular carcinomas (NID2, TM7SF2, TRIM2, and GLTSCR2). Comparative genomic groupings identified differentially expressed genes that may lead to better classification of follicular thyroid neoplasms. Such genes may be used in future prospective validation studies to establish clinically useful and complementary diagnostic markers.