Expression In Papillary Thyroid Carcinoma Research Articles

APOE expression in papillary thyroid carcinoma: Influencing tumor progression and macrophage polarization

BackgroundAs metastatic papillary thyroid carcinoma becomes increasingly challenging to treat, immunotherapy has emerged as a new research direction. Tumor-associated macrophages (TAMs) influence the occurrence, invasion, and metastasis of tumors. Apolipoprotein E (APOE) can regulate the polarization changes of macrophages and participate in the remodeling of the tumor microenvironment. However, the role of APOE in regulating the polarization and biological functions of TAMs in papillary thyroid carcinoma (PTC) remains unclear, as it acts as a dual biomarker. MethodsWe probed APOE expression in PTC tissues using immunohistochemical staining. A cell co-culture model was established where different APOE-expressing K1 cells were co-cultured with THP-1-derived M0 macrophages. An in-depth analysis of macrophage polarization behavior was performed using real-time quantitative polymerase chain reaction, enzyme-linked immunosorbent assay, and western blotting. Subsequently, the impact of APOE-regulated macrophages on tumor cell behavior, especially proliferation, migration, and invasion, was evaluated utilizing IncuCyte ZOOM system, flow cytometry, colony formation, and scratch experiments. Finally, we used a xenograft model to confirm the effects of APOE on PTC tumorigenesis. ResultsTumor dimensions, stage, and lymphatic metastases were significantly associated with increased APOE expression in PTC tissues. K1 cells were markedly limited in their proliferation, migration, and invasion abilities when APOE expression was silenced, a process mediated by the PI3K/Akt/NF-κB signaling axis. Moreover, APOE is a key facilitator of the enhancement of the anti-inflammatory cytokines IL-10 and TGF-β1. In PTC cellular models, APOE contributed to the phenotypic shift of THP-1 derived macrophages towards an M2 phenotypic polarization, predominantly through the modulation of IL-10. Furthermore, in vivo studies involving athymic nude mice have demonstrated pivotal role of APOE in tumor progression and the induction of M2-like TAM polarization. ConclusionOur results elucidated that APOE could promote the shift of TAMs from M0-type to M2-type polarization by regulating inflammatory factors expressions in K1 cell through the PI3K/Akt/NF-κB pathway. These findings are crucial for understanding the molecular mechanisms underlying PTC pathogenesis and for developing immunological drugs to treat this disease.

Clinicopathological significance of epidermal growth factor receptor expression in papillary thyroid carcinoma: a meta-analysis.

This study aimed to determine the relationship between the expression of epidermal growth factor receptor (EGFR) and pathological indicators in papillary thyroid carcinoma (PTC). PubMed, Embase, Web of Science, MEDLINE, and Cochrane Library databases were searched for relevant clinical trials. The odds ratio (OR) and 95% confidence interval (CI) showed the effect magnitude of the expression of EGFR, age, gender, tumor size, lymph node metastasis (LNM), extrathyroid extension, and tumor, lymph node, metastasis) stage. Stata 12.0 was used for statistical analysis of data. A total of 845 cases of PTC were included through the retrieval of 8 studies performed abroad. EGFR significantly correlated with extrathyroid extension (OR=3.25; 95% CI: 1.25-8.43; Z=2.42; P=0.015), LNM (OR=8.40; 95% CI: 5.44-12.97; Z=9.61; P=0.000), TNM stage (OR=2.30, 95% CI: 1.51-3.51; Z=3.87; P=0.000), and tumor size (OR=1.68; 95% CI: 1.06-2.68; Z=2.19; P=0.03). EGFR had no correlation with age (OR=1.13; 95% CI: 0.83-1.53; Z=0.77; P=0.44), gender (OR=0.93; 95% CI: 0.66-1.33; Z=0.38; P=0.70). Sensitivity analysis demonstrated that the studies by Cui Tang and Alfred King Yin Lam in LNM impacted the pooled OR. After removing these two studies, relatively stable results between the expression of EGFR and LNM were obtained. The results in the expression of EGFR is frequent and cancer-specific event in PTC. Besides, the expression of EGFR was involved in the progression and metastasis of PTC.

Bioinformatic analysis of CCND2 expression in papillary thyroid carcinoma and its impact on immune infiltration

To investigate cyclin D2 (CCND2) expression in papillary thyroid carcinoma (PTC) and its association with the clinicopathological features. The public databases TCGA, TIMER 2.0 and UALCAN were used to explore CCND2 expression level in PTC and adjacent tissues, and its diagnostic value for PTC was analyzed using ROC curves. GO enrichment analysis of CCND2-related differentially expressed genes (DEGs) in PTC was performed, and tumor immune infiltration of CCND2 in thyroid cancer was analyzed using TIMER database and CIBERSORT data source. RT-qPCR and Western blot were used to detect CCND2 expression in normal human thyroid cell line Nthy-ori-3-1 and human PTC cell lines TPC-1 and BCPAP. CCND2 expression was also detected in clinical specimens of PTC and adjacent tissues by immunohistochemistry, and its correlation with clinicopathological features of the patients were analyzed. Informatic analysis revealed significantly higher CCND2 mRNA expression in thyroid cancer than in the adjacent tissues (P < 0.001) in close correlation with tumor stage, gender, age, pathological subtype, and lymph node involvement (P < 0.05). ROC curve analysis showed that at the cutoff value of 4.983, the diagnostic sensitivity, specificity, and accuracy of CCND2 expression for PTC was 83.6%, 94.9%, and 78.5%, respectively. CCND2 expression was positively correlated with B cells, CD4+ T cells, and macrophages (P < 0.001) and negatively with CD8+ T cells (P < 0.01), and also correlated with memory B-cell infiltration, CD4+ T-cell memory activation, M2 macrophages, resting mast cells, and mast cell activation (P < 0.05). RT-qPCR, Western blot and immunohistochemistry showed significantly higher CCND2 expression in the PTC cells than in Nthy-ori-3-1 cells (P < 0.01) and also in clinical PTC tissues than in the adjacent tissues (P < 0.05) in correlation with tumor size, lymph node metastasis and TNM stage (P < 0.05). CCND2 overexpression is closely correlated with tumor progression and immune cell infiltration in PTC patients..

BRAF mutation, selected miRNAs and genes expression in primary papillary thyroid carcinomas and local lymph node metastases

Mutations in cancer-related genes are now known to be accompanied by epigenetic events in carcinogenesis by modification of the regulatory pathways and expression of genes involved in the pathobiology. Such cancer-related mutations, miRNAs and gene expression may be promising molecular markers of the most common papillary thyroid carcinoma (PTC). However, there are limited data on their relationships. The aim of this study was to analyse the interactions between BRAF mutations, selected microRNAs (miR-21, miR-34a, miR-146b, and miR-9) and the expression of selected genes (LGALS3, NKX2–1, TACSTD2, TPO) involved in the pathogenesis of PTC. The study cohort included 60 primary papillary thyroid carcinomas (PTC) that were classified as classical (PTC/C; n=50) and invasive follicular variant (PTC/F; n=10), and 40 paired lymph node metastases (LNM). BRAF mutation status in primary and recurrent/persistent papillary thyroid carcinomas was determined. The mutation results were compared both between primary and metastatic cancer tissue, and between BRAF mutation status and selected genes and miRNA expression in primary PTC. Furthermore, miRNAs and gene expression were compared between primary PTCs and non-neoplastic tissue, and local lymph node metastatic tumor, respectively. All studied markers showed several significant mutual interactions and contexts. In conclusion, to the best our knowledge, this is the first integrated study of BRAF mutational status, the expression levels of mRNAs of selected genes and miRNAs in primary PTC, and paired LNM.

Diagnostic and prognostic utility of TROP-2, SLP-2, and CXCL12 expression in papillary thyroid carcinoma.

Papillary thyroid carcinoma (PTC) is the most frequent thyroid malignancy. Histopathological examination is widely accepted as the gold standard test for the diagnosis of PTC. However, the histopathological examination sometimes can't differentiate PTC from other thyroid diseases. Differentiating PTC from other thyroid diseases is essential for a therapeutic approach and prognosis. The current study was performed to investigate the utility of TROP-2, SPL-2, and CXCL12 mRNA and protein expression in discriminating PTC from other thyroid diseases that mimic PTC. The current study was performed on 75 cases of surgically resected thyroid glands. The cases were distributed in two groups: the PTC group and the non-PTC group. The PTC group consisted of 35 cases (25 patients of the classic PTC variant and 10 patients of the PTC follicular variant). The non-PTC group consisted of 40 cases (10 cases were multinodular goiter, 5 cases were Graves' disease, 5 cases were Hashimoto thyroiditis, 15 patients were follicular adenoma (FA) and 5 cases were follicular carcinoma). TROP-2, SPL-2, and CXCL12 mRNA expression were estimated by qRT-PCR, and protein expression was estimated by immunohistochemistry. There were upregulated TROP-2, SPL-2, and CXCL12 mRNA and protein expressions in PTC compared to non-PTC (P< 0.001, for each). There was a statistically significant upregulation in the mRNA expression of the three genes among PTC cases with larger tumor sizes (P< 0.001, for each), those with tumor stages III and IV (P= 0.008, 0.002 and < 0.001 respectively), and those with LN metastasis (P< 0.001, for each). Moreover, there was a statistically significant upregulation in CXCL-12 gene expression among PTC cases with extra-thyroid extension (P< 0.001). mRNA expression of TROP-2, SPL-2, and CXCL12 among PTC cases increased in larger tumor size, tumor stages III and IV, and LN metastasis. Moreover, there was an increase in CXCL-12 gene expression among PTC cases with extra-thyroid extension. Thus, TROP-2, SPL-2, and CXCL12 expressions could be possible diagnostic and prognostic markers in PTC.

Chronic Lymphocytic Thyroiditis with Oncocytic Metaplasia Influences PD-L1 Expression in Papillary Thyroid Carcinoma.

Despite the increasing recognition of PD-L1 as predictor of immunotherapeutic response in various malignancies, its role and prognostic significance in thyroid cancer remain underexplored and subject to debate. This study begins to address this gap by comprehensively analyzing PD-L1 expression in papillary thyroid carcinoma (PTC) and investigating its correlation with key clinicopathological variables. We conducted immunohistochemistry (IHC) to assess PD-L1 expression in whole-tissue sections from 121 primary papillary thyroid carcinoma (PTC) cases. We then analyzed the correlations between PD-L1 expression and various clinicopathological variables. PD-L1 expression was detected in 33.1% of papillary thyroid carcinomas (PTCs), predominantly exhibiting weak to moderate intensity. Notably, this study found no significant correlation between PD-L1 expression and various clinicopathological variables. The lack of association with traditional factors such as age, sex, histological subtype, and tumor size suggests the complex and multifaceted nature of PD-L1 regulation in PTC. Multivariate logistic regression analysis identified chronic lymphocytic thyroiditis with oncocytic metaplasia as the sole independent predictor of PD-L1 expression (P = 0.014), underlining the potential influence of the tumor microenvironment on immune checkpoint expression in PTC. Our study underscores the intricate interplay between chronic lymphocytic thyroiditis with oncocytic metaplasia and PD-L1 expression in papillary thyroid carcinoma. The observed link suggests a potential avenue for therapeutic intervention using anti-PD-1/PD-L1 therapies in surgery-refractory PTC. Understanding the dynamics of immune checkpoint regulation in the context of the tumor microenvironment is crucial for devising effective treatment strategies. Future research endeavors should delve deeper into the molecular mechanisms underlying this interaction and explore its implications for patient outcomes. As the field of immunotherapy continues to evolve, our findings contribute valuable insights into the complex immunological landscape of thyroid cancer.

P53-associated miRNAs repress lncRNA ZFAS1 to retard the proliferation of papillary thyroid carcinoma.

Thyroid carcinoma is the most frequent malignancy among endocrine-related tumours. Papillary thyroid carcinoma (PTC) is the main type of thyroid carcinoma, and almost 80% cases of thyroid carcinoma are diagnosed as PTC. The molecular mechanism underlying PTC progression is unclear. This study aims to investigate the potential mechanisms of zinc finger antisense 1 (ZFAS1) function in PTC. The expression of ZFAS1 and p53 was determined by quantitative polymerase chain analysis (qPCR) in PTC tissues derived from 20 PTC patients. Quantitative chromatin immunoprecipitation assay (qChIP) analysis was performed to validate the target of ZFAS1/p53 and miRNAs/p53. The Gene Expression Omnibus (GEO) dataset GSE94908 was analysed to obtain the differentially expressed p53-associated microRNAs (miRNAs). Luciferase assay validated the target of ZFAS1/miRNAs, and 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to determine the cell proliferation. The expression of ZFAS1 was up-regulated in the tissues derived from PTC patients, and the expression of ZFAS1 was negatively associated with p53 expression in PTC. The expression of ZFAS1 was significantly higher in the MDA-T120 cells harbouring mutant p53. We validated that ZFAS1 is a direct target of p53. In PTC cells, p53 directly repressed the ZFAS1 expression. In addition, we determined that miR-135b-5p and miR-193a-3p are directly induced by p53 in PTC cells. Interestingly, p53-targeted miR-135b-5p, miR-193a-3p, and miR-34b repressed the expression of ZFAS1 via the seed-matching sequences in the 3'-untranslated region (3'-UTR) of ZFAS1, and thereby suppressed PTC cell proliferation induced by ZFAS1. The oncogenic lncRNA ZFAS1 is directly repressed by p53 in PTC. p53-mediated miRNAs including miR-135b 5p, miR-193a-3p, and miR-34b repress ZFAS1 expression, and thereby inhibit the proliferation of PTC.

CD73 mitigates ZEB1 expression in papillary thyroid carcinoma

BackgroundZEB1, a core transcription factor involved in epithelial-mesenchymal transition (EMT), is associated with aggressive cancer cell behavior, treatment resistance, and poor prognosis across various tumor types. Similarly, the expression and activity of CD73, an ectonucleotidase implicated in adenosine generation, is an important marker of tumor malignancy. Growing evidence suggests that EMT and the adenosinergic pathway are intricately linked and play a pivotal role in cancer development. Therefore, this study focuses on exploring the correlations between CD73 and ZEB1, considering their impact on tumor progression.MethodsWe employed CRISPR/Cas9 technology to silence CD73 expression in cell lines derived from papillary thyroid carcinoma. These same cells underwent lentiviral transduction of a reporter of ZEB1 non-coding RNA regulation. We conducted studies on cell migration using scratch assays and analyses of cellular speed and polarity. Additionally, we examined ZEB1 reporter expression through flow cytometry and immunocytochemistry, complemented by Western blot analysis for protein quantification. For further insights, we applied gene signatures representing different EMT states in an RNA-seq expression analysis of papillary thyroid carcinoma samples from The Cancer Genome Atlas.ResultsSilencing CD73 expression led to a reduction in ZEB1 non-coding RNA regulation reporter expression in a papillary thyroid carcinoma-derived cell line. Additionally, it also mitigated ZEB1 protein expression. Moreover, the expression of CD73 and ZEB1 was correlated with alterations in cell morphology characteristics crucial for cell migration, promoting an increase in cell polarity index and cell migration speed. RNA-seq analysis revealed higher expression of NT5E (CD73) in samples with BRAF mutations, accompanied by a prevalence of partial-EMT/hybrid state signature expression.ConclusionsCollectively, our findings suggest an association between CD73 expression and/or activity and the post-transcriptional regulation of ZEB1 by non-coding RNA, indicating a reduction in its absence. Further investigations are warranted to elucidate the relationship between CD73 and ZEB1, with the potential for targeting them as therapeutic alternatives for cancer treatment in the near future.

Clinical significance of matrix metalloproteinase-9 expression in papillary thyroid carcinoma: a meta-analysis

ObjectiveThe purpose of this study was to investigate the relationship between the expression of matrix metalloproteinase-9 (MMP-9) and pathological indexes in papillary thyroid carcinoma (PTC).Evidence obtainedThe database was searched in PubMed, Embase, CNKI, and Web of Science databases for relevant clinical trials. The odds ratio (OR) and 95% confidence interval (CI) show the effect of MMP-9 expression and age, tumour size, gender, lymph node metastasis (LNM), and TNM (tumour, lymph node, metastasis) stage. Statistical analysis of the data was performed using Stata 17.0.Evidence synthesisA total of 1433 patients with PTC were included in this meta-analysis. MMP-9 expression was significantly correlated with LNM (OR = 3.92, 95% CI = 2.71–5.65, P = 0.000), tumour size (OR = 1.69, 95% CI = 1.13–2.52, P = 0.011), and TNM stage (OR = 2.95, 95% CI = 2.10–4.13, P = 0.000), but not with gender (OR = 0.90, 95% CI = 0.66–1.22, P = 0.487) and age (OR = 1.36, 95% CI = 0.93–1.98, P = 0.115).ConclusionsOur meta-analysis showed that MMP-9 was significantly associated with LNM, tumour size, and TNM stage; therefore, MMP-9 may be a reliable prognostic biomarker for patients with PTC. However, more high-quality studies are needed to support these findings further.

Increased SOCS1 and SOCS3 expression in papillary thyroid carcinoma and its association with prognosis.

The majority of patients with papillary thyroid carcinoma (PTC) have good outcomes, although the identification of new predictors of a poor prognosis would be beneficial. Chronic thyroiditis is a precancerous condition in which proinflammatory cytokines enhance biologically aggressive features. This study investigated the expression of suppressor of cytokine signaling proteins (SOCS) 1 and 3, which are negative feedback inhibitors, in PTC and benign thyroid nodules (BTN), and analyzed the relations among biomarker expression, pathological prognosis, and clinical features. The pathological materials and clinical data of 100 patients with PTC and 40 with BTN were retrospectively analyzed. Immunohistochemical SOCS1 and SOCS3 staining were performed. Besides comparing SOCS1 and SOCS3 expression between PTC and BTN, we analyzed the expression according to pathological factors and clinical variables. The expression levels of the proteins were significantly higher in PTC than in BTN (p=0.001). SOCS1 expression was higher in older patients with PTC than in younger patients (p=0.001). Unlike SOCS1, SOCS3 was related to the risk group; these groups were distinguished based on the American Thyroid Association (ATA) risk stratification system (p=0.001). SOCS3 was also significantly related to lymph node involvement (p=0.007), capsule invasion (p=0.005), and extrathyroid extension (p=0.009). The increased SOCS1 and SOCS3 expression in PTC confirms their roles in thyroid carcinogenesis. Antibodies to both SOCS1 and SOCS3 might aid the diagnosis of PTC through immunohistological staining. SOCS3 provides information on lymph node status and aids risk stratification.

EPAS1/HIF-2α Acts as an Unanticipated Tumor-Suppressive Role in Papillary Thyroid Carcinoma.

Overexpression of hypoxia-inducible factors led to tumor angiogenesis and tumor progression. However, unlike HIF-1α, the role of EPAS1/HIF-2α in papillary thyroid carcinoma (PTC) was unknown. Here, we aimed to investigate the role of EPAS1/HIF-2α in PTC. EPAS1/HIF-2α expression of fresh frozen tumor samples and adjacent tissues in Tongji Hospital of 46 PTC patients was detected by RT-PCR. Gene expression datasets of PTC patients were gained from The Cancer Genome Atlas (TCGA) database. The Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) and gene set enrichment analysis (GSEA) were used to explore the potential biological function of EPAS1/HIF-2α. The effect of EPAS1/HIF-2α on immune microenvironment of PTC was analyzed in R package "estimate". The sensitivity to various targeted drugs was quantified in R package "pRRophetic", while the sensitivity to immunotherapy was estimated based on TCIA website. We found higher EPAS1/HIF-2α mRNA expression in PTC was associated with lower N stage, M stage, and better progression-free time (PFS) and disease-free time (DFS). Further, biological function analysis indicated that EPAS1/HIF-2α was mainly involved in PI3K-Akt signaling pathway. EPAS1/HIF-2α expression was positively related with CD8+ T cell infiltration and negatively related to PD-L1 expression and tumor mutation burden. Patients with low EPAS1/HIF-2α expression were more than likely to get a profit from Sorafenib, Dabrafenib, Cetuximab, Bosutinib, and immune checkpoint blockade. Our results suggested that EPAS1/HIF-2α played an unanticipated tumor-suppressive role in PTC. EPAS1/HIF-2α contributed to anti-tumor immunity by promoting CD8+ T cell infiltration and inhibiting PD-L1 expression in PTC.

Nrf2 as a novel diagnostic biomarker for papillary thyroid carcinoma

Papillary thyroid carcinoma (PTC) is the most common thyroid malignancy. However, it is very difficult to distinguish PTC from benign carcinoma. Thus, specific diagnostic biomarkers are actively pursued. Previous studies observed that Nrf2 was highly expressed in PTC. Based on this research, we hypothesized that Nrf2 may serve as a novel specific diagnostic biomarker. A single-center retrospective study, including 60 patients with PTC and 60 patients with nodular goiter, who underwent thyroidectomy at the Central Theater General Hospital from 2018 to July 2020, was conducted. The clinical data of the patients were collected. Nrf2, BRAF V600E, CK-19, and Gal-3 proteins were compared from paraffin samples of the patients. Through this study, we obtained the following results: i) Nrf2 exhibits high abundance expression in PTC, but not in adjacent to PTC and nodular goiter; increased Nrf2 expression could serve as a valuable biomarker for PTC diagnosis; the sensitivity and specificity for the diagnosis of PTC were 96.70% and 89.40%, respectively. ii) Nrf2 also shows higher expression in PTC with lymph node metastasis, but not adjacent to PTC and nodular goiter, thus the increased Nrf2 expression might serve as a valuable predictor for lymph node metastasis in PTC patients; the sensitivity and specificity for the prediction in lymph node metastasis were 96.00% and 88.57%, respectively; excellent diagnostic agreements were found between Nrf2 and other routine parameters including HO-1, NQO1 and BRAF V600E. iii) The downstream molecular expression of Nrf2 including HO-1 and NQO1 consistently increased. In conclusion, Nrf2 displays a high abundance expression in human PTC, which leads to the higher expression of downstream transcriptional proteins: HO-1 and NQO1. Moreover, Nrf2 can be used as an extra biomarker for differential diagnosis of PTC and a predictive biomarker for lymph node metastasis of PTC.

YAP1 and P53 Expression in Papillary Thyroid Carcinoma.

One of the most prevalent endocrine system cancers is papillary thyroid carcinoma, with complicated predisposing factors and pathogenesis. YAP1 (Yes-associated protein 1) is a well-known oncogene; its activity is increased in a variety of human malignancies and has recently been paid great attention. The present study examines YAP1 and P53 immunohistochemical expression in papillary thyroid carcinoma and investigates the association of their expression with the available clinicopathological risk factors to assess their possible prognostic role. The current study used paraffin blocks of 60 cases of papillary thyroid carcinoma, which were immunohistochemically assessed for YAP1 and p53 expression. The study examined the association of their expression with clinicopathological characteristics. YAP1 expression was observed in 70% of papillary thyroid carcinoma cases. A statistically significant relation was observed between YAP1 expression and tumor size, tumor stage, tumor focality, lymph node metastases, and extrathyroidal extension (P-values were =0.003, > 0.001, 0.037, 0.025, and 0.006), respectively. p53 expression was observed in 85% of papillary thyroid carcinoma cases. A statistically significant relation was observed between p53 expression and tumor size (P=0.001) and tumor stage (P>0.001). A statistically significant relation was noticed between YAP1 and P53 expression (P=0.009). YAP1 expression was found to be associated with many high-risk clinicopathological characteristics in patients with papillary thyroid carcinoma and with p53 expression; thus, it seems that YAP1 may have a specific impact on the patient's outcome.

Potential significance ofpodoplanin immunohistochemical expression in papillary thyroid carcinoma.

Podoplanin (PDPN) is alymphatic endothelial marker expressed by arange ofhuman malignancies in which it has been shown to contribute to tumor progression and metastasis. However, there is alack ofthestudies, examining thefunction ofPDPN in thyroid cancer. Thecurrent study was performed to explore the possible diagnostic value ofPDPN expression in papillary thyroid cancer (PTC) and to evaluate themarker's potential for prediction ofregional lymph node metastasis. Lymphatic vascular density (LVD) and thestromal/cancer-associated fibroblasts (CAFs), labeled by PDPN, were examined in PTC compared to theother thyroid lesions. Thecurrent study included 50 cases ofPTC and 50 cases ofnon-PTC thyroid lesions. Immunohistochemical staining was performed using monoclonal PDPN antibodies. Podoplanin expression was scored as positive and negative. Podoplanin expression was found in 36% ofPTC cases, but it was not found in benign, low risk (borderline), or malignant lesions other than PTC. Furthermore, lymph node metastasis was significantly correlated with PDPN expression, LVD and CAFs (p-values<0.00001,<0.001 and 0.0002 respectively). These findings support thediagnostic utility ofPDPN expression in PTC and its predictive value for LN metastasis.

The expression of stromal biomarkers in small papillary thyroid carcinomas

BackgroundThe importance of stroma for tumor progression is recognized for many cancer types. In this study, we aim to evaluate the expression of types I (Col1) and IV (Col4) collagens, alpha-smooth muscle actin (a-SMA), and matrix metallopeptidase 9 (MMP-9) in the tumor stroma of small papillary thyroid carcinoma (PTC).Material and methodsTwenty-five non-metastatic small PTCs (pT1N0) and nineteen metastatic small PTCs (pT1N1b) including corresponding metastatic lateral lymph nodes were selected and paraffinized tissue blocks retrieved. The samples were stained for Col1, COL4, a-SMA, and MMP-9 antibodies using immunohistochemistry. The expression of the stromal proteins was scored and analyzed based on the location, intensity, and distribution.ResultsCol1 and Col4 expression were significantly higher in normal thyroid tissue compared to PTC tissue. On the contrary, expression of a-SMA and MMP-9 was higher in PTC tissue compared to normal thyroid tissue. Both Col1 and Col4 were significantly more highly expressed in the non-metastatic tumors compared with metastatic tumors. The expression of a-SMA and MMP9 was slightly, but not significantly, higher in the metastasized tumors and their respective lymph nodes. There was a significant correlation between the metastasized tumors and their respective lymph nodes in Col1 and MMP-9 expression.ConclusionsCol1, Col4, a-SMA, and MMP-9 expression in PTCs differs significantly from that of normal thyroid tissue. The higher expression of Col1 and Col4 in normal thyroid tissue and in the non-metastasized tumors indicates that Col1 and 4 might have a potential protective role in tumor progression. The higher expression of a-SMA and MMP9 in PTCs indicates that these proteins might have a role in promoting PTC progression and aggressiveness.

Diagnostic Significance of FNAB miRNA Expression in Papillary Thyroid Carcinoma.

The aim of the study was to evaluate the diagnostic utility of specific miRNAs in the preoperative assessment of thyroid nodules. One hundred and sixty thyroid fine needle aspiration biopsy (FNAB) samples with suspected thyroid carcinoma were collected. To detect the levels of miRNA expression in FNAB, next generation small RNA sequencing was performed in 60 samples. Based on the results obtained, three miRNAs (miR125A, miR200B, miR4324) were selected for further analysis. Based on the most frequently reported miRNAs in the literature associated with thyroid papillary carcinoma (PTC), two more miRNA (miR146B, miR221) were selected for further validation, using real-time reverse transcriptase polymerase chain reaction (RT-PCR) in 36 benign and 64 PTC samples. Expression of miR125A, miR146B, miR221, and miR4324 was significantly higher in patients with PTC compared with benign thyroid nodules (p ˂ 0.05). miR125A and miR4324 were also significantly more highly expressed in patients with extrathyroidal tumor extension compared to those without extrathyroidal PTC extension (p < 0.001). We also found a significantly higher expression of miR221 (p = 0.043) in patients with multifocal carcinomas compared to patients with single foci carcinomas. This prospective study showed that the expression analysis of four miRNAs (miR125A, miR146B, miR221, and miR4324) improve accuracy of FNAB, which could allow a better pre-operative diagnostic and prognostic assessment of thyroid malignancies.

Correlation between myoferlin expression and lymph node metastasis in papillary thyroid carcinoma.

BackgroundMyoferlin is a multifunctional protein expressed in various normal and cancer cells, with novel oncogenic roles being newly discovered. Recently, correlations have been found between myoferlin expression and unfavorable prognosis in various carcinomas. This study investigated the prognostic role of myoferlin expression in papillary thyroid carcinoma (PTC), specifically that associated with nodal metastasis.MethodsWe collected clinicopathological data and PTC tissues from 116 patients who had been admitted to Gyeongsang National University Hospital in 2010. Immunohistochemical analysis was performed on surgical specimen-derived tissue microarray blocks. Myoferlin expression was graded, and the relationship between expression level and pathological features of tumors based on the American Joint Committee on Cancer staging system was evaluated.ResultsOf the 116 patient samples, 100 cases exhibited positive myoferlin expression. Higher grade of myoferlin expression was correlated with lower T category group (p = .010). Presence of lymph node metastasis was determined to be significantly correlated with low-grade myoferlin expression (p = .019), with no significant difference between pN1a and pN1b tumors.ConclusionsOur study revealed an adverse correlation between myoferlin expression and pathological features of PTC, evidence of the potential prognostic role of myoferlin in PTC lymph node metastasis.

CD56 Expression in Papillary Thyroid Carcinoma Is Highly Dependent on the Histologic Subtype: A Potential Diagnostic Pitfall.

Loss of CD56 expression has been regarded as a diagnostic marker of papillary thyroid carcinoma (PTC). However, certain variants of PTC can aberrantly express CD56. Using a digital image analysis tool, we evaluated H-scores of CD56 immunostaining in 216 thyroid tumors. The H-score of the CD56 of all PTCs was lower than that of noninvasive follicular thyroid neoplasm with papillary-like nuclear features (NIFTP) (P<0.001). The H-scores of CD56 were lower in classic PTC, the infiltrative follicular variant, and the diffuse sclerosing variant than in other PTC variants (P<0.001), whereas the H-scores were higher in tall cell variant, Warthin-like variant, and cribriform-morular variant than in classic PTC (P<0.001). The optimal cutoff value of H-scores for the CD56 expression was 180 for differentiating the NIFTP from the follicular adenoma and 30 for the differential diagnosis of NIFTP and infiltrative follicular variant PTC. CD56 expression is predominantly lost in classic and infiltrative follicular variants of PTCs and more preserved in the other histologic subtypes of PTC and NIFTP. CD56 is particularly useful for differentiating PTC from follicular-pattern thyroid neoplasms, but the aberrant expression in uncommon variants of PTC could be a diagnostic pitfall.

Evaluation of ITGA3 as a Biomarker of Progression and Recurrence in Papillary Thyroid Carcinoma.

ObjectiveTo investigate the expression of ITGA3 and its association with clinical outcomes in papillary thyroid carcinoma (PTC).MethodsThe expression level, association with clinicopathologic characteristics, co-expressed genes, signaling pathways of ITGA3 in thyroid cancer were comprehensively analyzed using bioinformatics analysis through multiple public gene databases. PTC specimens and cell lines were used to verify the results of bioinformatics analysis.ResultsData mining based on the Oncomine database revealed that ITGA3 expression in classical PTC and tall cell variant PTC was much higher than that in normal thyroid tissue except the follicular variant PTC. Analysis based on The Cancer Genome Atlas (TCGA) database showed that the expression of ITGA3 varies greatly in pathological stages, pathological types, tumor invasion stages, and lymph node metastasis stages of thyroid carcinoma. High expression level of ITGA3 was correlated with tumor regional invasion and lymph node metastasis. Multivariate analysis using logistic regression model showed that high expression of ITGA3 was a risk factor that associated with PTC recurrence and lymph node metastasis. Survival analysis showed that patients with high expression of ITGA3 in PTC had a poorer relapse-free survival (RFS) than patients with low expression of ITGA3 (P < 0.05). Immunohistochemistry experiments showed that the expression of ITGA3 in recurrent thyroid cancer tissues was stronger than that in no-recurrent thyroid cancer tissues (P < 0.05). Knockdown of ITGA3 by sh-RNA in PTC cell lines suppresses cell viability and invasive and migrating capacity.ConclusionITGA3 is overexpressed in PTC, especially in those with higher tumor invasion grades and lymph node metastasis, and was associated with recurrence and poor RFS of PTC. High expression of ITGA3 may have the potential role of predicting PTC recurrence and lymph node metastasis.

Expression of epithelial membrane protein (EMP) 1, EMP 2, and EMP 3 in thyroid cancer.

Purpose Epithelial membrane protein (EMP) 1, EMP2, and EMP3 are expressed in various types of tumors and have been reported to be involved in carcinogenesis. In this study, we aimed to investigate the expression of these proteins in primary and metastatic thyroid cancer and its clinical implication. Methods EMP1, EMP2, and EMP3 immunohistochemistry was performed using tissue microarrays of 545 primary thyroid carcinomas [338 papillary thyroid carcinoma (PTC), 111 follicular carcinoma (FC), 69 medullary carcinoma (MC), 23 poorly differentiated carcinoma (PDC), and 4 anaplastic carcinoma (AC)] and 59 recurrent or metastatic PTCs. Results EMP1 showed high expression in AC, PTC, FC (P<0.001), and EMP2 was highly expressed in AC (P<0.001). EMP1 and EMP2 were not expressed in stromal cells. Expression of EMP3 in tumor cells [EMP3 (T)] was higher in PDC, PTC, and AC (P<0.001), and expression in stromal cells [EMP3 (S)] was observed only in AC and PTC (P=0.001). The expression of EMP1 (P=0.002) and EMP3 (T) (P<0.001) was higher in conventional PTC than in follicular variant PTC. PTC with BRAF V600E mutation showed higher expression of EMP1 (P<0.001), EMP3 (T) (P<0.001), and EMP3 (S) (P=0.012) than PTC without BRAF V600E mutation. In the PTC without BRAF V600E mutation group, expression of EMP3 (S) was associated with shorter disease free survival (P=0.004). Metastatic PTC showed higher EMP2 (3.4% vs. 0%, P=0.022) and lower EMP3 (T) (44.1% vs. 66%, P=0.001) than primary PTC. Conclusions Expression of EMP1, EMP2, and EMP3 is different according to the subtypes of thyroid cancer. Further studies are needed to determine their role as prognostic markers and treatment target in thyroid cancer.