Thyroid Tissue Specimens Research Articles

High Expression of Immune Checkpoint Molecules in Different Types of Thyroid Cancer.

This study aimed to investigate the expression of programmed cell death protein-1 (PD-1) and its ligand (PD-L1) immune checkpoint molecules in thyroid carcinomas and determine their association with the clinicopathological characteristics of patients. Thyroid tissue specimens from100 patients diagnosed with primary thyroid carcinomas including papillary thyroid carcinoma (PTC), follicular thyroid carcinoma (FTC), medullary thyroid carcinoma (MTC), and anaplastic thyroid carcinoma (ATC) were collected. Sections were prepared from formalin-fixed paraffin-embedded samples, and PD-1 and PD-L1 expressions were examined using immunohistochemistry. PD-1 was detected in tumor-infiltrating lymphocytes (TILs) in 88% of the patients and tumor cells in 28% of the patients with 10% in PTC, 5% in FTC, 5% in MTC, and 8% in ATC). PD-L1 was found in tumor cells and TILs in 30% and 79% of the patients, respectively. Moreover, a significant difference was observed in PD-1 and PD-L1 expression between tumor cells and TILs across different tumor types. However, their expression in tumor cells and TILs was significantly higher in ATC compared to other tumor types. Additionally, the expression of PD-1 and PD-L1 was significantly associated with an advanced stage, higher tumor size, tumor necrosis, and mitosis. A significant positive correlation was also observed between the expression of PD-1 and PD-L1 in tumor cells and TILs. The higher expression of PD-1 and PD-L1 may contribute to tumor progression. Therefore, combinational immunotherapy by these immune checkpoint inhibitors might be a promising strategy for clinical improvement in patients with thyroid cancer, especially those with ATC.

Jagged 1 Regulates The Proliferation and Metastasis of Human MDA-T68 Thyroid Cancer Cells.

The current study evaluated the expression profile and explored the therapeutic implications of Jagged 1 in human thyroid cancer. This experimental study was conducted in 60 paired specimens of papillary thyroid and adjacent normal tissues. Gene expression was determined by quantitative real time polymerase chain reaction (qRT-PCR) and western blotting. Transfection of cancer cells was performed by using Lipofectamine 2000. The cell proliferation of PTC cells was estimated by MTT assay. Clonogenic assay was performed for analysis of colony forming potential of cancer cells. The apoptosis of PTC cells was studied by using AO/EB and Annexin V-FITC/PI staining methods. Flow cytometry was done to analyze the cell cycle phase distribution of cancer cells. Migration and invasion PTC cells were determined respectively with the wound-healing and transwell assays. The impact of Jagged 1 silencing was investigated in vivo in a xenografted mice model followed by Immunohistochemistry (IHC) analysis. We found significant (P<0.05) upregulation of Jagged 1 in human thyroid cancer. Silencing of Jagged 1 caused significant (P<0.05) reduction in proliferation and colony formation of MDA-T68 cells. The inhibitory effects of Jagged 1 silencing were found to be due to the induction of apoptosis. We also found enhancement of Bax and repression of Bcl-2 protein levels in MDA-T68 cells. Wound healing assay indicated significant (P<0.05) inhibition of cell migration of MDA-T68 thyroid cancer cells. Additionally, we found that invasion of the thyroid cancer cells was reduced by 55% upon silencing of Jagged 1. Moreover, Jagged 1 silencing was found to cause inhibition of the Notch intracellular domain (NICD) and Notch target gene, Hes-1 expression. Finally, Jagged 1 silencing inhibited the xenografted tumors in vivo. The findings suggest that Jagged 1 regulates the development of thyroid cancer that may act as a therapeutic target for managing thyroid cancer.

Whole-exome sequencing and bioinformatic analyses revealed differences in gene mutation profiles in papillary thyroid cancer patients with and without benign thyroid goitre background.

Papillary thyroid cancer (PTC) is the most common thyroid malignancy. Concurrent presence of cytomorphological benign thyroid goitre (BTG) and PTC lesion is often detected. Aberrant protein profiles were previously reported in patients with and without BTG cytomorphological background. This study aimed to evaluate gene mutation profiles to further understand the molecular mechanism underlying BTG, PTC without BTG background and PTC with BTG background. Patients were grouped according to the histopathological examination results: (i) BTG patients (n = 9), (ii) PTC patients without BTG background (PTCa, n = 8), and (iii) PTC patients with BTG background (PTCb, n = 5). Whole-exome sequencing (WES) was performed on genomic DNA extracted from thyroid tissue specimens. Nonsynonymous and splice-site variants with MAF of ≤ 1% in the 1000 Genomes Project were subjected to principal component analysis (PCA). PTC-specific SNVs were filtered against OncoKB and COSMIC while novel SNVs were screened through dbSNP and COSMIC databases. Functional impacts of the SNVs were predicted using PolyPhen-2 and SIFT. Protein-protein interaction (PPI) enrichment of the tumour-related genes was analysed using Metascape and MCODE algorithm. PCA plots showed distinctive SNV profiles among the three groups. OncoKB and COSMIC database screening identified 36 tumour-related genes including BRCA2 and FANCD2 in all groups. BRAF and 19 additional genes were found only in PTCa and PTCb. "Pathways in cancer", "DNA repair" and "Fanconi anaemia pathway" were among the top networks shared by all groups. However, signalling pathways related to tyrosine kinases were the most significantly enriched in PTCa while "Jak-STAT signalling pathway" and "Notch signalling pathway" were the only significantly enriched in PTCb. Ten SNVs were PTC-specific of which two were novel; DCTN1 c.2786C>G (p.Ala929Gly) and TRRAP c.8735G>C (p.Ser2912Thr). Four out of the ten SNVs were unique to PTCa. Distinctive gene mutation patterns detected in this study corroborated the previous protein profile findings. We hypothesised that the PTCa and PTCb subtypes differed in the underlying molecular mechanisms involving tyrosine kinase, Jak-STAT and Notch signalling pathways. The potential applications of the SNVs in differentiating the benign from the PTC subtypes requires further validation in a larger sample size.

High Prevalence of Common Human Viruses in Thyroid Tissue.

IntroductionEvidence points to viral infections as possible triggers of autoimmune thyroid disease (AITD), but little is known about the prevalence of common viruses in the thyroid gland. Using a novel approach based on virus enrichment in multiple cell lines followed by detection of the viral genome and visualization of viral proteins, we investigated the presence of multiple human viruses in thyroid tissue from AITD patients and controls.MethodsThyroid tissue was collected by core needle biopsy or during thyroid surgery from 35 patients with AITD (20 Graves’ disease and 15 Hashimoto’s thyroiditis). Eighteen thyroid tissue specimens from patients undergoing neck surgery for reasons other than thyroid autoimmunity served as controls. Specimens were tested for the presence of ten different viruses. Enteroviruses and human herpesvirus 6 were enriched in cell culture before detection by PCR and immunofluorescence, while the remaining viruses were detected by PCR of biopsied tissue.ResultsForty of 53 cases (75%) carried an infectious virus. Notably, 43% of all cases had a single virus, whereas 32% were coinfected by two or more virus types. An enterovirus was found in 27/53 cases (51%), human herpesvirus 6 in 16/53 cases (30%) and parvovirus B19 in 12/53 cases (22%). Epstein-Barr virus and cytomegalovirus were found in a few cases only. Of five gastroenteric virus groups examined, only one was detected in a single specimen. Virus distribution was not statistically different between AITD cases and controls.ConclusionCommon human viruses are highly prevalent in the thyroid gland. This is the first study in which multiple viral agents have been explored in thyroid. It remains to be established whether the detected viruses represent causal agents, possible cofactors or simple bystanders.

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.

A five-gene panel refines differential diagnosis of thyroid nodules.

BackgroundMolecular testing for oncogenic mutations in fine‐needle aspiration has showed high predictive value in identifying malignant lesions from thyroid nodules with indeterminate cytology.MethodsTo figure out an efficient and economical gene panel for most medical institutions in China, we designed a five‐gene panel including BRAF/NRAS/KRAS/HRAS/TERT genes and conducted a retrospective study to evaluate the role of this five‐gene diagnostic panel in differential diagnosis of thyroid nodules.ResultsA total of 665 patients with 695 thyroid nodules were investigated in the current study. The fine‐needle aspiration biopsy and surgically separated thyroid tissue specimens were harvested to test BRAF, TERT, NRAS, KRAS, and HRAS mutations. We identified 261 mutations in 665 patients, including 177 V600E mutations in BRAF. Three hundred and sixty‐nine patients who underwent thyroid surgery after completion of the initial clinical and cytological evaluation were enrolled in the final analysis. The diagnostic sensitivity, specificity, and accuracy of the combination of FNAB cytology and five‐gene detection were 74.7%, 93.8%, and 84.8%, respectively. BRAF V600E and five‐gene panel could recognize 46.4% and 53.6% of papillary thyroid carcinoma in the patients with cytologically indeterminate nodules.ConclusionThe five‐gene panel can effectively improve the sensitivity, negative predictive value, and accuracy of fine‐needle aspiration biopsy cytology, especially in the patients with cytologically indeterminate nodules.

Detection of Human Papillomavirus in Papillary Thyroid Carcinoma and its Association with Tumor Staging and Pathologic Features.

Background: The role of human papillomavirus (HPV), as a common infection, has been evaluated in many cancers such as the cervix and squamous cell carcinoma of the head and neck. To the best of our knowledge, for the first time, the association of HPV with papillary thyroid carcinoma (PTC) and its pathologic features are investigated.Methods: A retrospective cross-sectional study was conducted from May 2014 to January 2018 in several hospitals affiliated to Shiraz University of Medical Sciences, Shiraz, Iran. Thyroid tissue specimens of patients diagnosed with PTC (n=82) and benign thyroid nodules (n=77) were collected using the consecutive sampling method. The presence of HPV in PTC, adjacent normal tissue, and benign thyroid nodules was evaluated using the polymerase chain reaction (PCR) method. The frequency of HPV positivity in PTC tissues was compared with benign thyroid nodules and adjacent normal tissue. Association of pathologic features of PTC with HPV positivity was also investigated. Data were analyzed using SPSS version 21.0, and P values less than 0.05 were considered statistically significant.Results: HPV PCR positivity was observed in 3.8% of benign thyroid nodules and 13.4% of PTC samples but in none of the adjacent normal tissues. After adjustment for age and sex, the prevalence of HPV PCR positivity in the PTC tissues was significantly more than the benign thyroid nodules (P=0.015). The prevalence was also significantly higher than the adjacent normal tissues (P<0.001).Conclusion: There was a significant association between PTC and HPV positivity. Further studies are required to determine the cause and effect of the association between these two conditions.

Additional Tissue Sampling Trials Did Not Change Our Thyroid Practice.

Simple SummarySome studies have suggested that the use of additional tissue blocks to diagnose follicular thyroid carcinoma (FTC) could increase the accuracy of diagnosis and improve prognosis, and entire capsule sampling was recommended for a definitive diagnosis of borderline thyroid tumors in 2016. We conducted a study in 2016 to examine whether additional tissue sampling of the encapsulated follicular tumor increases the diagnosis of malignant cases in our patient cohort. Furthermore, the diagnosis was reclassified according to the 4th edition of the World Health Organization’s classification system. The additional tissue sampling only had a slight impact on our thyroid practice and resulted in no benefits to the patient; therefore, we decided to cease it.This study aimed to determine whether additional tissue sampling of encapsulated thyroid nodules would increase the frequency of follicular thyroid carcinoma (FTC) diagnoses. We examined thyroid tissue specimens from 86 patients suspected of FTC (84.9% female; mean age, 49.0 ± 17.8 years). The number of tissue blocks created for pathological assessments ranged from 3 to 20 (mean, 9.1 ± 4.1); the numbers in the previous method recommended by the Japanese General Rules for the Description of Thyroid Cancer and additional blocks ranged from 1 to 12 (mean, 6.0 ± 2.8) and from 1 to 8 (mean, 3.1 ± 2.0), respectively. The additional blocks were subsequently examined to determine whether any diagnoses changed from those based on the previous method. Five patients were diagnosed with FTC using the previous method; however, additional tissue blocks led to the diagnosis of FTC in 6 patients, as 1 diagnosis was revised from follicular adenoma to FTC. It has been reported that increasing the number of tissue blocks used for pathological assessments can increase the frequency of FTC diagnoses; however, this was not clinically significant in thyroid carcinoma, which requires completion thyroidectomy and radioactive iodine treatment. It resulted in no benefits to the patient because all minimally invasive FTCs, follicular tumors of uncertain malignant potential (FT-UMP), and follicular adenomas are treated with lobectomy alone in Japan. Additional tissue sampling only had a slight impact on our thyroid practice; therefore, we decided to cease it.

Fatty Acid Metabolism as a Potential Therapeutic Target in Thyroid Carcinoma

Background: Papillary thyroid carcinoma (PTC) accounts for the majority of thyroid cancer and affects a large number of individuals. The pathogenesis of PTC has not been completely elucidated thus far. Metabolic reprogramming is a common feature in tumours. Our previous research revealed the reprogramming of lipid metabolism in PTC. Further studies on lipid metabolism reprogramming may help elucidate the pathogenesis of PTC. Methods: Clinical samples of PTC and para-tumour tissue were analysed using lipidomic, proteomic, and metabolomic approaches. A multi-omics integrative strategy was adopted to identify the important pathways in PTC. The findings were further confirmed using western blotting, tissue microarray, and bioinformatics. Findings: The differentially expressed substances included 146 lipids, 2975 proteins, and 217 metabolites in PTC samples. Multi-omics data and the results of integrated analysis revealed that the three steps of fatty acid metabolism (hydrolysis, transportation, and oxidation) were significantly enhanced in PTC. The key enzymes in the three steps, namely LPL, FATP2, and CPT1A, respectively, were overexpressed. Moreover, their expression was strongly associated with the TNM stage, lymph node metastasis, and PTC prognosis. In addition, the levels of several types of phospholipids and sphingolipids increased significantly in PTC. Interpretation: Our data suggested that enhanced fatty acid metabolism supplied additional energy and substrates for tumour growth, invasion, and metastasis in PTC. This may help elucidate the mechanism underlying PTC pathogenesis and identify the potential therapeutic targets for PTC. Funding Information: Work in the authors' laboratory is supported by grants from National Natural Science Foundation of China (No. 81702647, NO. 81972819), Taishan Scholar Foundation of Shandong Province (No. tspd20181201), Collaborative Innovation Center of Technology and Equipment for Biological Diagnosis and Therapy in Universities of Shandong, Natural Science Foundation of Shandong Province (No. ZR201702190483, No. ZR2020YQ57), Key Technology Research and Development Program of Shandong (No. 2016GSF201161). Declaration of Interests: The authors declare no competing interests. Ethics Approval Statement: This prospective study was approved by the institutional ethical committee of Qilu Hospital of Shandong University, Shandong Province, China. All the specimens were collected in the Qilu Hospital of Shandong University. The participants provided written informed consent for the use of thyroid tissue specimens for research.

Immunohistochemical Expression of Extracellular Matrix Protein-1 in Neoplastic Thyroid Lesions

Background: Upregulated Extracellular matrix protein 1 (ECM 1) expression has been observed in several malignancies including breast, thyroid carcinoma, cutaneous melanoma and gastric carcinoma. ECM 1 expression has been evaluated in various epithelial malignancies using real time – reverse transcriptase polymerase chain reaction (RT-PCR). The purpose of current study is to evaluate the immunohistochemical expression of ECM 1 in various neoplastic lesions of thyroid.&#x0D; &#x0D; Methods: The study included 81 neoplastic thyroid tissue specimens (21 benign and 60 malignant lesions) received over a period of 5 years (2012-2017). Hematoxylin and Eosin stained sections of the tissue specimens were carefully examined under light microscope in order to establish the histopathological diagnosis. The immunohistochemical marker ECM 1 was applied on appropriate sections and the slides were evaluated accordingly.&#x0D; &#x0D; Result: The expression of ECM 1 was nil (grade 0, 47.62% cases) or weak (1+, 52.38% cases) in benign lesions while majority of malignant lesions (78.33%) showed strong homogenous and crisp staining (3+). The difference in the expression of ECM 1 between benign and malignant lesions was found to be statistically significant (p &lt; 0.001). ECM 1 has a diagnostic accuracy of 0.82 and sensitivity of 100% in differentiating benign from malignant lesions.&#x0D; &#x0D; Conclusion: ECM 1 is uniformly upregulated in all malignant thyroid neoplasms irrespective of the histological type of malignancy. Immunohistochemical expression of ECM 1 shows comparable results to RT-PCR. ECM 1 immunohistochemistry can thus serve as an important standalone marker of malignant thyroid tissue.

Application of HMWCK, CK19 and Galectin-3 as Immuno Histochemical Markers in the differential diagnosis of thyroid lesions

Introduction: Thyroid neoplasms constitute the most common of all endocrine neoplasms occurring worldwide and more than 95% arise from follicular epithelial cells. Thyroid tumors are more common in developed countries. The incidence of thyroid tumors has increased in past two decades.Aim: to assess the role of Galectin-3 in differentiating benign and malignant thyroid lesions along and combined use of CK-19 and HMW-CK in the differential diagnosis between papillary carcinoma and follicular carcinoma.Material and methods: The present study was conducted in 50 specimens of thyroid tissue received in the Department of Pathology, Government Medical College, Amritsar, after approval from the Institutional Ethics Committee. IHC staining was done using Galectin-3, CK-19 and HMW-CK immuno Histochemical markers. The IHC score was calculated by combining an estimate of the percentage of immunoreactive cells (quantity score) with an estimate of the staining intensity (staining intensity score).Results: Incidence of thyroid nodules most common in the age group between 20-40 years comprising about 66% of the total cases followed by 41-60 years. Females constituted about 74% (37 cases) in contrast to males with 13 cases comprising 26%. Prevalence of benign Lesions of thyroid was slightly higher 29 (58%) in comparison to malignant lesions which comprised of 21 (42%) of the study sample. Galactein 3 is found to be the most sensitive marker while HMWCK is found to be the most specific marker in distinguishing malignant thyroid lesions from benign thyroid lesions in present study. CK-19 and HMW CK when combined together in absence of Galactein 3 shows 100% sensitivity and high specificity in diagnosing malignant thyroid lesions. These combinations show increased specificity and positive predictive value compared to Galactein 3 when used alone.Conclusion: Therefore we can say that these immuno histochemical panels can be a useful means to increases the likelihood of detecting malignant tumors. These markers along with histopathological diagnosis can aid us in correct diagnosis and thus further help to optimize the management of patients with thyroid nodules and reduce unnecessary surgical resection of benign nodules.

Analytical validation of a novel targeted next-generation sequencing assay for mutation detection in thyroid nodule aspirates and tissue.

The identification of somatic mutations in cancer specimens enables detection of molecular markers for personalized treatment. We recently developed a novel molecular assay and evaluated its clinical performance as an ancillary molecular method for indeterminate thyroid nodule cytology. Herein we describe the analytical validation of the novel targeted next-generation sequencing (NGS) assay in thyroid samples from different sources. We present validation data of a novel NGS-based panel on 463 thyroid samples, including 310 fine-needle aspiration (FNA) specimens from different sources (FNA collected in preservative solution, liquid-based, and stained smears), 10 fresh frozen, and 143 formalin-fixed paraffin-embedded (FFPE) thyroid tissue specimens. Sequencing performance in the different samples was evaluated along with reproducibility, repeatability, minimum nucleic acid input to detect variants, and analytical sensitivity of the assay. All thyroid samples achieved high sequencing performance, with a mean base coverage depth ranging from 2228 × (in liquid-based FNA) to 3661 × (in FNA stained smears), and coverage uniformity ranging from 86% (in FFPE) to 95% (in FNA collected in preservative solution), with all target regions covered above the minimum depth required to call a variant (500×). The minimum nucleic acid input was 1 ng. Analytic sensitivity for mutation detection was 2-5% mutant allele frequency. This validation study of a novel NGS-based assay for thyroid nodules demonstrated that the assay can be reliably used on multiple thyroid sample types, including FNA from different sources and FF and FFPE thyroid samples, thus providing a robust and reliable assay to genotype thyroid nodules, which may improve thyroid cancer diagnosis and care.

Automatic cancer tissue detection using multispectral photoacoustic imaging.

In the case of multispecimen study to locate cancer regions, such as in thyroidectomy and prostatectomy, a significant labor-intensive processing is required at a high cost. Pathology diagnosis is usually done by a pathologist observing tissue-stained glass slide under a microscope. Multispectral photoacoustic (MPA) specimen imaging has proven successful in differentiating photoacoustic (PA) signal characteristics between a histopathology-defined cancer region and normal tissue. This is mainly due to its ability to efficiently map oxyhemoglobin and deoxyhemoglobin contents from MPA images and key features for cancer detection. A fully automated deep learning algorithm is purposed, which learns to detect the presence of malignant tissue in freshly excised ex vivo human thyroid and prostate tissue specimens using the three-dimensional MPA dataset. The proposed automated deep learning model consisted of the convolutional neural network architecture, which extracts spatially colocated features, and a softmax function, which detects thyroid and prostate cancer tissue at once. This is one of the first deep learning models, to the best of our knowledge, to detect the presence of cancer in excised thyroid and prostate tissue of humans at once based on PA imaging. The area under the curve (AUC) was used as a metric to evaluate the predictive performance of the classifier. The proposed model detected the cancer tissue with the AUC of 0.96, which is very promising. This model is an improvement over the previous work using machine learning and deep learning algorithms. This model may have immediate application in cancer screening of the numerous sliced specimens that result from thyroidectomy and prostatectomy. Since the instrument that was used to capture the ex vivo PA images is now being developed for in vivo use, this model may also prove to be a starting point for in vivo PA image analysis for cancer diagnosis.

Validation of Reference Genes for Normalization of Relative qRT-PCR Studies in Papillary Thyroid Carcinoma

Quantitative reverse transcription polymerase chain reaction (qRT-PCR) in thyroid tumors require accurate data normalization, however, there are no sufficient studies addressing the suitable reference genes for gene expression analysis in malignant and normal thyroid tissue specimens. The purpose of this study was to identify valid internal control genes for normalization of relative qRT-PCR studies in human papillary thyroid carcinoma tissue samples. The expression characteristics of 12 candidate reference genes (GAPDH, ACTB, HPRT1, TBP, B2M, PPIA, 18SrRNA, HMBS, GUSB, PGK1, RPLP0, and PGM1) were assessed by qRT-PCR in 45 thyroid tissue samples (15 papillary thyroid carcinoma, 15 paired normal tissues and 15 multinodular goiters). These twelve candidate reference genes were selected by a systematic literature search. GeNorm, NormFinder, and BestKeeper statistical algorithms were applied to determine the most stable reference genes. The three algorithms were in agreement in identifying GUSB and HPRT1 as the most stably expressed genes in all thyroid tumors investigated. According to the NormFinder software, the pair of genes including ‘GUSB and HPRT1’ or ‘GUSB and HMBS’ or ‘GUSB and PGM1’ were the best combinations for selection of pair reference genes. The optimal number of genes required for reliable normalization of qPCR data in thyroid tissues would be three according to calculations made by GeNorm algorithm. These results suggest that GUSB and HPRT1 are promising reference genes for normalization of relative qRT-PCR studies in papillary thyroid carcinoma.

Steroid receptor coactivator-1 interacts with NF-κB to increase VEGFC levels in human thyroid cancer.

Thyroid cancer is the most common endocrine cancer, and has a high incidence of lymphatic metastasis. Vascular endothelial growth factor C (VEGFC) is essential for development of lymphatic vessels and lymphatic metastases during carcinogenesis. Steroid receptor coactivator-1 (SRC-1) interacts with nuclear receptors and transcription factors to promote tumor proliferation and metastasis. However, the correlation between SRC-1 and VEGFC levels in the lymphatic metastases of thyroid cancer remains unclear. We analyzed 20-paired specimens of thyroid cancer tissue and normal thyroid tissue and found increased levels of SRC-1 and VEGFC proteins in 13/20 and 15/20 thyroid cancer specimens, respectively, when compared with those levels in specimens of normal thyroid tissue. A high level of SRC-1 expression was positively correlated with VEGFC and lymphatic endothelial cell marker LYVE-1 expression. Papillary thyroid carcinoma cell line TPC-1 displayed high levels of SRC-1 and VEGFC expression and was selected for stable knockdown of SRC-1 in vitro. Inhibition of SRC-1 significantly reduced the VEGFC levels in TPC-1 cells. We found that SRC-1 binds to transcription factor NF-kB (p50/p65), and that this coactivation complex directly promoted VEGFC transcription, which could be abrogated by SRC-1 knockdown. Up-regulated NF-kB signaling was also confirmed in thyroid cancer tissues. In vivo studies showed that SRC-1 knockdown restricted tumor growth, reduced the numbers of LYVE-1-positive lymphatic vessels, and decreased the levels of VEGFC in tumor tissues. These results suggest a tumorigenic role for SRC-1 in thyroid cancer via its ability to regulate VEGFC expression.

An immunohistochemical study of CTHRC1,Vimentin,E-cadherin expression in papillary thyroid carcinoma

Objective:To investigate the expression and significance of CTHRC1 in patients with papillary thyroid cancinoma.Method:We collected 63 specimens of papillary thyroid carcinoma tissue with 36 specimens of the adjacent normal thyroid tissue and 23 specimens of thyroid nodular goiter. The expressions of CTHRC1 protein in these tissues were detected by immunohistochemical staining. The correlations between the expressions of CTHRC1 with clinicopathologic features,E-cadherin and Vimentin expression were analyzed.Result:CTHRC1 protein expression levels in papillary thyroid carcinoma tissue were significantly higher than those in adjacent normal thyroid tissue and benign disease(P<0.01);CTHRC1 expression was significantly correlated with lymph node metastases(P<0.05). Moreover,CTHRC1 expression was correlated with the expression of E-cadherin and Vimentin(P<0.01).Conclusion:CTHRC1 is related with the occurrence and miligant transformation of papillary thyroid cancinoma. We speculated that CTHRC1 might play a role in the epithelial-mesenchymal transition of papillary thyroid cancinoma.

Discrimination of papillary thyroid cancer from non-cancerous thyroid tissue based on lipid profiling by mass spectrometry imaging.

The distinction of papillary thyroid carcinomas from benign thyroid lesions has important implication for clinical man-agement. Classification based on histopathological features can be supported by molecular biomarkers, including lipidomic signatures, identified with the use of high-throughput mass spectrometry techniques. Formalin fixation is a standard procedure for stabilization and preservation of tissue samples, therefore this type of samples constitute highly valuable source of clinical material for retrospective molecular studies. In this study we used mass spectrometry imaging to detect lipids discriminating papillary cancer from not cancerous thyroid directly in formalin-fixed tissue sections. For this purpose imaging and profiling of lipids present in non-malignant and cancerous thyroid tissue specimens were conducted. High resolution MALDI-Q-Ion Mobility-TOF-MS technique was used for lipidomic analysis of formalin fixed thyroid tissue samples. Lipids were identified by the comparison of the exact molecular masses and fragmentation pathways of the protonated molecule ions, recorded during the MS/MS experiments, with LIPID MAPS database. Several phosphatidylcholines (32:0, 32:1, 34:1 and 36:3), sphingomyelins (34:1 and 36:1) and phosphatidic acids (36:2 and 36:3) were detected and their abundances were significantly higher in cancerous tissue compared to non-cancerous tissue. The same lipid species were detected in formalin-fixed as in fresh-frozen tissue, but [M + Na]+ ions were the most abundant in formalin fixed whereas [M + K]+ ions were predominant in fresh tissue. Our results prove the viability of MALDI-MSI for analysis of lipid distribution directly in formalin-fixed tissue, and the potential for their use in the classification of thyroid diseases.

ST6GalNAcII mediates tumor invasion through PI3K/Akt/NF-κB signaling pathway in follicular thyroid carcinoma.

Altered sialylation, closely associated with tumor progression and metastasis, has been implicated in human thyroid carcinoma. The present study investigated the alteration in expression of ST6GalNAcII involved in invasion and to clarify the possible mechanism of ST6GalNAcII in the metastasis process in human follicular thyroid carcinoma cell lines. Using real-time PCR, western blot and IHC analysis, ST6GalNAcII differed in three follicular thyroid cancer cell lines (FTC133, primary and FTC238, lung metastasis). It also showed differential expression in follicular thyroid carcinoma and tissue specimens. In addition, we analyzed the PI3K/Akt signaling pathway. The altered expression of ST6GalNAcII corresponded to changed invasive phenotype of FTC-238 and FTC-133 cells in vitro and in vivo. Further studies showed that regulating ST6GalNAcII expression markedly modulated the activity of the phosphoinositide 3-kinase (PI3K)/Akt signaling pathway. Targeting the PI3K/Akt pathway by its specific inhibitor LY294002, or by Akt small interfering RNA (siRNA) resulted in reduced capacity in invasion of FTC-238. In conclusion, taken together, our results imply that ST6GalNAcII activated the invasion in follicular thyroid cancer cells through regulating the activity of PI3K/Akt pathway.

Multicompartment metabolism in papillary thyroid cancer.

In many cancers, varying regions within the tumor are often phenotypically heterogeneous, including their metabolic phenotype. Further, tumor regions can be metabolically compartmentalized, with metabolites transferred between compartments. When present, this metabolic coupling can promote aggressive behavior. Tumor metabolism in papillary thyroid cancer (PTC) is poorly characterized. Immunohistochemical staining of tissue samples. Papillary thyroid cancer specimens from 46 patients with (n = 19) and without advanced disease (n = 27) were compared to noncancerous thyroid tissue (NCT) and benign thyroid specimens (n = 6 follicular adenoma [FA] and n = 5 nodular goiter [NG]). Advanced disease was defined as the presence of lateral neck lymphadenopathy. Immunohistochemistry was performed for translocase of outer mitochondrial membrane 20 (TOMM20), a marker of oxidative phosphorylation, and monocarboxylate transporter 4 (MCT4), a marker of glycolysis. Papillary thyroid cancer and FA thyrocytes had high staining for TOMM20 compared to NCT and nodular goiter (NG) (P < 0.01). High MCT4 staining in fibroblasts was more common in PTC with advanced disease than in any other tissue type studied (P < 0.01). High MCT4 staining was found in all 19 cases of PTC with advanced disease, in 11 of 19 samples with low-stage disease, in one of five samples of FA, in one of 34 NCT, and in 0 of six NG samples. Low fibroblast MCT4 staining in PTC correlated with the absence of clinical adenopathy (P = 0.028); the absence of extrathyroidal extension (P = 0.004); low American Thyroid Association risk (P = 0.001); low AGES (age, grade, extent, size) score (P = 0.004); and low age, metastasis, extent of disease, size risk (P = 0.002). This study suggests that multiple metabolic compartments exist in PTC, and low fibroblast MCT4 may be a biomarker of indolent disease. N/A. Laryngoscope, 126:2410-2418, 2016.

New insight into ectopic thyroid glands between the neck and maxillofacial region from a 42-case study.

BackgroundEctopic thyroid is a rare disease. In the present study at the 9th People’s Hospital in Shanghai, China, 42 patients’ ectopic thyroid glands between the neck and maxillofacial region were subjected to a retrospective and transverse study based on data from 1978 to 2012 to explore the natural characteristics of ectopic thyroid.MethodsThe patients’ clinical data were collected. In addition, scintigraphy (Tc-99 m, Iodine-131), CT scan, histology and pathology were performed. The protein expression of thyroid transcription factor-1 (TTF-1), thyroglobulin (TG), calcitonin (CT), Ki-67 and parathyroid hormone (PTH) were analyzed from paraffin wax-stored specimens of ectopic thyroid tissue compared with those of orthotopic thyroid tissue.ResultsThere were 42 total ectopic thyroid patients, approximately 1.24 patients per year on average at our hospital. These patients were aged from 6 to 85 years old, and there were 35 females (83.3 %), seven males (16.7 %). In total, 27 of the patients had lingual thyroid (64 %); seven, sublingual thyroid (17 %); five, dual areas occupied by ectopic thyroid (12 %) and three, other types (7 %). The following conditions were also presented: nodular goiter (13 %), adenoma (8.7 %) and Hashimoto’s thyroiditis (4.3 %), no malignancy and no accompanying ectopic parathyroid. TTF-1 expression was significantly higher in ectopic samples than that in orthotopic samples (P = 0.007), but CT and Ki-67 levels displayed no difference. PTH was negative in ectopic tissue.ConclusionEctopic thyroid is a rare disease and females were more prone to the disease. The most frequent location was lingual thyroid. Nodular goiter, adenoma and Hashimoto’s thyroiditis was observed as orthotopic thyroid without accompanying ectopic parathyroid. TTF-1 was highly expressed in ectopic tissue, which may be related to abnormal embryogenesis leading to the thyroid gland being in an abnormal position. The expression of calcitonin (CT) and Ki-67 was not increased, and there were no malignant cells in any sample, which could indicate that it is not easy for ectopic thyroids to become malignant between the neck and maxillofacial region.