Characterization of a novel four-miRNA signature in papillary thyroid carcinoma: Integrating molecular profiling, hormonal regulation, and diagnostic implications in populations with rising PTC incidence.

Objective:To evaluate the level of melatonin and the role of melatonin in the metastasis of papillary thyroid carcinoma in patients with papillary thyroid carcinoma. Method:We measured serum melatonin levels in 81 patients with papillary thyroid carcinoma（PTC） ,20 patients with multinodular goiter（MNG） and 20 healthy adults using ELISA. The relationship between melatonin and clinicopathological features of PTC were analyzed.The expression of MT1 and MT2 in two subtypes of melatonin receptor in 81 cases of papillary thyroid carcinoma and adjacent tissues were detected by immunohistochemical SP method, and its the mean optical density（MOD） image was analyzed by Image Pro Plusversion（IPP） image processing software. Result:Serum melatonin concentration in patients with PTC was significantly higher than that in MNG patients and normal controls（P<0.05）. The level of melatonin in the primary tumor T≥2 cm group was significantly higher than that in the T<2 cm group. Patients with positive cervical lymph nodes（N≥1） had significantly higher melatonin levels than lymph node negatives（N=0）（P<0.05）. The MT1 and MT2 receptors were expressed in both PTC and paracancerous tissues, mainly in the cell membrane and cytoplasm. The expression of MT1 receptor was low in the two groups, and there was no statistical difference. The expression of MT2 receptor in PTC tissues Significantly higher than the adjacent tissues（P<0.05）, further studies showed that the expression of MT2 receptor in PTC tissues was associated with cervical lymph node metastasis, and the expression of MT2 receptor in PTC tissues with cervical lymph node metastasis was significantly lower than that without metastasis （P<0.05）. Conclusion:Serum melatonin levels in PTC patients were higher than those in MNG and control groups, which may be associated with low malignancy of PTC; melatonin inhibits PTC metastasis, which exerts anti-PTC metastasis mainly through MT2 receptors.

The association of the human leucocyte antigen (HLA) system with thyroid carcinomas is not clear. We sought to relate HLA alleles to susceptibility to papillary thyroid carcinoma (PTC) and also to clinical and pathological characteristics of PTC patients. The distribution of HLA in 181 unrelated Caucasian patients with PTC was compared to the HLA distribution in 315 normal controls, 31 patients with follicular carcinoma (FTC), 29 patients with lymphocytic thyroiditis (LT) and 50 patients with multinodular goitre (MNG), using a microlymphocytotoxicity assay. Compared to normal controls, patients with PTC showed a significantly increased frequency of HLA-DQ4 [12.8%vs. 3.5%, P=0.0005, P(corrected) (P(c))=0.0032, odds ratio (OR)=4.058, 95% confidence interval (95% CI)=1.820-9.045] and HLA-DR8 (10.9%vs. 4.3%, P=0.013, P(c) > 0.05, OR=2.752, 95% CI=1.275-5.940). DQ4 and DR8 were also significantly increased in patients with MNG (DQ4, 16.3%; DR8, 16.3%) compared to controls (DQ4, P=0.0019, P(c)=0.011, OR=5.420, 95% CI=1.978-14.852; DR8, P=0.0044, P(c)=0.062). Linkage disequilibrium (LD) for these two alleles was present in controls (D=0.0130, P=9.7e-57) and in MNG patients (D=0.0730, P=4.6e-19) but not in PTC patients (D=0.038, P>0.05). In the subgroup of PTC subjects with concomitant 0thyroidal neoplasias (n=27), there was a significant (P< 0.05) increase in the frequency of B57 (18.5%), DR11 (56.5%) and DQ3 (81.8%) compared to PTC patients without coexistent neoplasias (2.0%, 21% and 47%, respectively). No significant differences of HLA allele distribution was found in relation to PTC histology, age at diagnosis (> 45 or <or45 years), gender or tumour-node-metastasis (TNM) staging. In patients with FTC, the frequency of DR17 (FTC=51.6%; controls =; P=0.0009; P(c)=0.0138) was significantly increased compared to controls. Patients with LT showed a higher frequency of the DR11 allele (48.3%) than controls (DR11=21.3%; P=0.0028, P(c)=0.039, OR=3.445, 95% CI=1.568-7.567). We have typed the largest series of patients with thyroid carcinomas reported to date, and found that DR8 and DQ4 are independent susceptibility markers for PTC.

BackgroundDespite the presence of evidence that establishes a strong correlation between oxidative stress and thyroid cancer, there exists a scarcity of research that investigates the specific role of glutathione as an important antioxidant in this particular context. The objective of this study was to assess the altered balance of oxidative stress in cases of thyroid cancer, which includes both papillary thyroid carcinoma (PTC) and micro PTC (mPTC), by examining and comparing the total antioxidant capacity (TAC), total oxidant status (TOS), oxidative stress index (OSI), reduced glutathione (GSH), oxidized glutathione (GSSG), and GSSG/GSH ratio with those of individuals diagnosed with multinodular goiter (MNG) as well as Healthy subjects.Materials and methodsPlasma samples were collected from 92 patients (23 mPTC, 23 PTC, 23 MNG, 23 Healthy). The levels of TAC, TOS, GSH, and GSSG were measured using a commercial assay kits, and the OSI and GSSG/GSH ratio were calculated for each sample. Statistical analyses were performed to compare the oxidative stress between the groups.ResultsThe plasma levels of TOS were significantly higher in the mPTC, PTC, and MNG groups compared to the Healthy individuals (p < 0.05). The OSI in the mPTC and PTC groups showed a significant increase compared to the Healthy group (p < 0.05). The levels of GSH in mPTC and PTC were markedly lower compared to the Healthy subjects (p < 0.01). Interestingly, the concentration of GSH in mPTC was found to be considerably lower than in PTC and MNG patients (p < 0.01).ConclusionThese findings indicate that GSH may be a useful biomarker for evaluating oxidative stress and antioxidant system status in patients with PTC, especially mPTC. Low levels of GSH may indicate increased levels of oxidative stress, which may contribute to the development and progression of mPTC to PTC.

Regulatory T cells (Treg) suppress the immune reaction. The aim of the present study was to investigate the clinicopathologic significance and roles of Treg in papillary thyroid carcinoma (PTC) patients with and without Hashimoto's thyroiditis. Flow cytometry was used to detect the percentage of CD4+CD25+CD127low/- Treg among CD4+ T cells in peripheral blood. FoxP3+ Treg were detected by immunohistochemistry in the tumor tissues. The percentage of CD4+CD25+CD127low/- Treg among CD4+ T cells was significantly higher in PTC patients than that in multinodular goiter (MNG) patients. There were large numbers of tumor-infiltrating FoxP3+ Treg in primary PTC and metastatic lymph nodes tissues; however, there was no FoxP3 expression in the MNG tissues. Higher percentage of Treg both in peripheral blood and tumor tissues was associated with extrathyroidal extension and lymph nodes metastasis. The percentage of CD4+CD25+CD127low/- Treg among CD4+ T cells in peripheral blood of PTC patients with Hashimoto's thyroiditis (HT) was significantly lower, whereas the infiltration of FoxP3+ Treg in tissues of PTC with HT tended to be increased. We concluded that the percentage of Treg increased in peripheral blood as well as in the tumor tissues of PTC patients compared with that of MNG patients. The high percentage of Treg was associated with aggressiveness. There may be a compensatory expansion of Treg at the sites of inflammation in tissues of PTC with HT contributing to the immune response suppression.

Fine needle aspiration (FNA) biopsy is an established procedure by which to sample thyroid nodules to ascertain etiology and produce a diagnosis conveying risk of malignancy with recommended patient follow-up. This procedure is well-tolerated and endorsed given the accessibility and vascularity of the thyroid gland. FNA cytopathology has proven efficacious for the primary assessment of thyroid nodules. Well-differentiated papillary thyroid carcinoma (PTC) and benign lymphocytic (Hashimoto) thyroiditis (HT) are distinct thyroid lesions that may be reported with diagnostic confidence based on their characteristic cytomorphologic features. However depending on the adequacy of FNA sampling and the morphology of aspirated cellular material, thyroid nodules with coexisting PTC and HT may pose diagnostic pitfalls. This may be dependent upon: (a) the architectural nature of the coexisting lesions in-vivo; (b) whether both lesions are adequately sampled through FNA; and (c) which of the cell types and cytomorpholo...

The genetic factors that determine the risk of papillary thyroid carcinoma (PTC) among patients with multinodular goiter (MNG) remain undefined. Because thyroid transcription factor-1 (TTF-1) is important to thyroid development, we evaluated whether the gene that encodes it, TITF-1/NKX2.1, is a genetic determinant of MNG/PTC predisposition. Twenty unrelated PTC patients with a history of MNG (MNG/PTC), 284 PTC patients without a history of MNG (PTC), and 349 healthy control subjects were screened for germline mutation(s) in TITF-1/NKX2.1 by sequencing of amplified DNA from blood. The effects of the mutation on the growth and differentiation of thyroid cells were demonstrated by ectopic expression of wild-type (WT) and mutant proteins in PCCL3 normal rat thyroid cells, followed by tests of cell proliferation, activation of cell growth pathways, and transcription of TTF-1 target genes. All statistical tests were two-sided. A missense mutation (1016C>T) was identified in TITF-1/NKX2.1 that led to a mutant TTF-1 protein (A339V) in four of the 20 MNG/PTC patients (20%). These patients developed substantially more advanced tumors than MNG/PTC or PTC patients without the mutation (P = .022, Fisher exact test). Notably, this germline mutation was dominantly inherited in two families, with some members bearing the mutation affected with MNG, associated with either PTC or colon cancer. The mutation encoding the A339V substitution was not found among the 349 healthy control subjects nor among the 284 PTC patients who had no history of MNG. Overexpression of A339V TTF-1 in PCCL3 cells, as compared with overexpression of WT TTF-1, was associated with increased cell proliferation including thyrotropin-independent growth (average A339V proliferation rate = 134.27%, WT rate = 104.43%, difference = 34.3%, 95% confidence interval = 12.0% to 47.7%, P = .010), enhanced STAT3 activation, and impaired transcription of the thyroid-specific genes Tg, TSH-R, and Pax-8. This is the first germline mutation identified in MNG/PTC patients. It could contribute to predisposition for MNG and/or PTC and to the pathogenesis of PTC.

BackgroundGalectin-3 is a member of the beta-galactoside-binding protein family and functions as a modulator of cell growth through galactoside-binding protein correlated with the occurrence and metastasis of papillary thyroid carcinoma (PTC).MethodsA systematic review of published articles on Web of Science and PubMed was performed. After establishing inclusion and exclusion criteria, nine articles were selected. Three studies referred to galectin-3 expression in PTC and non-PTC patients. Three studies referred to galectin-3 expression in PTC patients with lymph node metastasis (LNM) and without LNM. Three studies referred to galectin-3 expression in both PTC (with and without LNM) and non-PTC patients. Data analysis was performed by using RevMan5.2 software.ResultsA total of 424 patients from six eligible studies that provided data about galectin-3 expression in PTC and non-PTC patients were included. A total of 378 patients from six eligible studies that provided data about galectin-3 expression in PTC with LNM and without LNM were included. Immunohistochemistry technique was used in all the studies. Galectin-3 was found to be a highly sensitive (275/424, 64.86%) marker in the diagnosis of PTC, but was found to be expressed only in a few cases involving other types of thyroid lesions (58/424, 13.68%). The odds ratio, expressed as PTC group versus other thyroid lesions group, was 13.97 (95% CI: 7.51–26.01, P<0.00001). The results also showed that the positive expression rates of galectin-3 in PTC patients with LNM were higher than those in PTC patients without LNM.ConclusionThis meta-analysis demonstrated that galectin-3 may become a potentially useful immunomarker to distinguish between PTC and non-PTC patients. In addition, PTC patients with positive expression of galectin-3 were more prone to LNM.

Based on strong research evidence, thyroid nodules in children and teenagers are more likely to be malignant than in adults. Based on strong research evidence, a history of ionizing radiation to the head or neck is an independent risk factor for the development of thyroid malignancies. There is strong research evidence, including a recent meta-analysis, supporting the use of fine-needle aspiration biopsy in the evaluation of all pediatric and adolescent patients presenting with a thyroid nodule. The surgical management and postoperative care of pediatric and adolescent patients who have well-differentiated thyroid carcinomas remains controversial, because the rarity of the disease limits the ability to conduct randomized, prospective research studies. Numerous studies have demonstrated that, despite presenting with more advanced disease, pediatric and adolescent patients with thyroid carcinoma have a higher survival rate than adults. The American Thyroid Association has issued strong evidence-based recommendations for the management of medullary thyroid carcinoma, including RET mutation testing and early prophylactic total thyroidectomy in children with high-risk mutations.

Epidemiology of Thyroid Cancer.

BackgroundThis study aimed to investigate the incidence and subtype of thyroid cancer in multinodular goitre (MNG) patients who underwent total thyroidectomy.MethodologyA cross-sectional study was conducted at the Khyber Teaching Hospital, screening 207 MNG patients who received complete thyroidectomies between July and December 2022. The senior consultant diagnosed thyroid cancer based on a complete history, physical examination, and laboratory and radiological studies. Ultrasound-guided fine-needle aspiration cytology was performed by a senior consultant radiologist. Bethesda categories for all lesions were recorded. All patients underwent thyroidectomy, and the diagnosis of thyroid cancer was confirmed on histopathology. ResultsA total of 207 patients were included in the study, with a mean age of 45.55 ± 8.75 years. Out of 207 patients, 24 (11.59%) were diagnosed with thyroid cancer. Out of 62 male patients, 15 (7.25%) had thyroid cancer. Out of 145 female patients, only nine had cancer (p < 0.001). Nine patients with thyroid cancer had a body mass index (BMI) below 18, compared to only five patients with a BMI of more than 30 kg/m2. The difference in age distribution was not significant in our study (p = 0.102). Conclusion In conclusion, our study sheds light on the frequency and potential risk factors associated with thyroid cancer in patients with multinodular goiter. Our findings reveal that papillary thyroid carcinoma is the most commonly observed form of thyroid cancer in this patient population, with around 12 percent of patients diagnosed with thyroid cancer. Notably, our study highlights that male patients and those with a lower BMI may have a greater risk of developing thyroid cancer in the context of multinodular goiter. The findings of this study have important implications for the care and follow-up of MNG patients who receive total thyroidectomy. Further research is needed to investigate the type and prognosis of thyroid cancer in patients with MNG.

Papillary thyroid carcinoma (PTC) is the fastest-growing disease caused by numerous molecular alterations in addition to previously reported DNA mutations. There is a compelling need to identify novel transcriptomic alterations that are associated with the pathogenesis of PTC with potential diagnostic and prognostic implications.Methods: We gathered and compared 242 expression profiles between paired PTC and adjacent normal tissues and identified and validated the coding and long non-coding RNAs (lncRNAs) associated with the extrathyroidal extension (ETE) of 655 PTC patients in two independent cohorts, followed by predicting their interactions with drugs. Co-expression, RNA interaction, Kaplan-Meier survival and multivariate Cox proportional regression analyses were performed to identify dysregulated lncRNAs and genes that correlated with clinical outcomes of PTC. Alternative splicing (AS), RNA circularization, and editing were also compared between transcriptomes to expand the repertoire of molecular alterations in PTC.Results: Numerous genes related to cellular microenvironment and steroid hormone response were associated with the ETE of PTC. Drug susceptibility predictions of the expression signature revealed two highly ranked compounds, 6-bromoindirubin-3'-oxime and lovastatin. Co-expression and RNA interaction analysis revealed the essential role of lncRNAs in PTC pathogenesis by modulating extracellular matrix and cell adhesion. Eight genes and two novel lncRNAs were identified that correlated with the aggressive nature and disease-free survival of PTC. Furthermore, this study provided the transcriptome-wide landscape of circRNAs in PTC and uncovered dissimilar expression profiles among circRNAs originating from the same host gene, suggesting the functional complexity of circRNAs in PTC carcinogenesis. The newly identified AS events in the SERPINA1 and FN1 genes may improve the sensitivity and specificity of these diagnostic biomarkers.Conclusions: Our study uncovered a comprehensive transcriptomic signature associated with the carcinogenesis and aggressive behavior of PTC, as well as presents a catalog of 10 potential biomarkers, which would facilitate PTC prognosis and development of new therapeutic strategies for this cancer.

BACKGROUND Papillary thyroid carcinoma is the most common malignant neoplasm of the thyroid gland, and fine-needle aspiration biopsy (FNAB) often is the initial diagnostic method used in its detection. Prior studies have shown that immunohistochemical staining for various cytokeratins in general, and cytokeratin 19 (CK19) in particular, can be applied as an ancillary technique for diagnosing papillary thyroid carcinoma in histologic specimens. In the current study the authors assessed the diagnostic utility of CK19 to detect papillary carcinoma effectively in cytologic preparations of thyroid FNABs. METHODS Immunocytochemical staining with CK19 was performed on cytologic aspirates from 37 papillary thyroid carcinomas and 36 other lesions of the thyroid (14 follicular adenomas, 10 multinodular goiters, 5 cases of Hashimoto thyroiditis, 6 oncocytic [Hürthle cell] neoplasms, and 1 follicular carcinoma). All cases included in the study had a corresponding histopathology specimen. RESULTS Positive immunocytochemical reactivity for CK19 was identified in 34 of 37 papillary carcinomas and in 1 of 36 other thyroid lesions (sensitivity of 92% and specificity of 97%). Although the strongest reactivity was obtained in methanol fixed thin layer preparations, the antibody also was effective in detecting papillary carcinoma in alcohol fixed and air-dried smears. The single false-positive case was a follicular adenoma with focal areas of papillary hyperplasia. All other aspirates including those from cases of Hashimoto thyroiditis, multinodular goiter, follicular adenoma, oncocytic neoplasms, and follicular carcinoma were negative. CONCLUSIONS CK19 is an effective, highly sensitive, and specific ancillary tool for the diagnosis of papillary carcinoma in thyroid FNABs. Cancer (Cancer Cytopathol) 2000;90:307–11. © 2000 American Cancer Society.

Papillary thyroid carcinoma is the most common malignant neoplasm of the thyroid gland, and fine-needle aspiration biopsy (FNAB) often is the initial diagnostic method used in its detection. Prior studies have shown that immunohistochemical staining for various cytokeratins in general, and cytokeratin 19 (CK19) in particular, can be applied as an ancillary technique for diagnosing papillary thyroid carcinoma in histologic specimens. In the current study the authors assessed the diagnostic utility of CK19 to detect papillary carcinoma effectively in cytologic preparations of thyroid FNABs. Immunocytochemical staining with CK19 was performed on cytologic aspirates from 37 papillary thyroid carcinomas and 36 other lesions of the thyroid (14 follicular adenomas, 10 multinodular goiters, 5 cases of Hashimoto thyroiditis, 6 oncocytic [Hürthle cell] neoplasms, and 1 follicular carcinoma). All cases included in the study had a corresponding histopathology specimen. Positive immunocytochemical reactivity for CK19 was identified in 34 of 37 papillary carcinomas and in 1 of 36 other thyroid lesions (sensitivity of 92% and specificity of 97%). Although the strongest reactivity was obtained in methanol fixed thin layer preparations, the antibody also was effective in detecting papillary carcinoma in alcohol fixed and air-dried smears. The single false-positive case was a follicular adenoma with focal areas of papillary hyperplasia. All other aspirates including those from cases of Hashimoto thyroiditis, multinodular goiter, follicular adenoma, oncocytic neoplasms, and follicular carcinoma were negative. CK19 is an effective, highly sensitive, and specific ancillary tool for the diagnosis of papillary carcinoma in thyroid FNABs.

Background:Epigenetic alterations such as DNA methylation are known as the main cause of different types of cancers through inactivation of tumor suppressor genes, especially thyroid cancer. Identification of novel and effective markers are important in diagnosis and prevention of thyroid cancer. In the present study, the expression and methylation of Solute carrier family 5 member 8 (SLC5A8) in Papillary Thyroid Carcinoma (PTC) in comparison to multinodular goiter (MNG) have been studied.Methods:Overall, 41 patients with PTC and 36 patients affected by MNG were recruited from four hospitals in Tehran and Qazvin, Iran in 2018. Thyroid tissues were obtained during thyroidectomy. RNA and DNA were extracted from thyroid tissues. Quantitative RT-PCR assay was performed for determining the mRNA level of SLC5A8 while Methylation-Sensitive High-Resolution Methylation was applied for assessing the methylation status.Results:Methylation status of three regions composed of 52 CpG islands in the promoter of SLC5A8 gene was studied by HRM assay. SLC5A8 level in PTC tissues was significantly downregulated in average 0.4 fold in comparison with MNG tissues (P=0.05). The aberrant methylation of SLC5A8 (b) region was remarkably different in PTC and MNG cases. The promoter methylation of SLC5A8 (c) was significantly related to BRAF mutations and vascular invasion in PTC patients.Conclusion:The aberrant promoter hyper methylation of SLC5A8 was related to aggressive PTC. Therefore, there is some evidence to support the hypothesis that SLC5A8 could be a paly important role in the development of PTC.

Hereditable predisposition to papillary thyroid carcinoma (PTC) and multinodular goiter (MNG) without evidence of an association with other malignancies as a distinct entity was recognized only recently. A meta-review of the literature on familial PTC (FPTC) was undertaken, and characteristics of families with frequent occurrence of PTC or MNG (or both) were summarized. A database on thyroid cancer patients maintained in our institution was searched for potential FPTC families. Clinical examinations were performed in 6 of 12 Hannover kindreds identified, and blood samples of all family members were collected for genetic analyses. Clinical presentations and histopathologic features of the FPTC cases were compiled. Based on the FPTC meta-review and own experience, predictive criteria to identify families at risk were developed: Exclusion criteria were previous radiation exposure and coincidence with neoplasia syndromes. Primary criteria for susceptibility to FPTC are (1) PTC in two or more first-degree relatives and (2) MNG in at least three first- or second-degree relatives of a PTC patient. Secondary criteria are diagnosis in a patient younger than 33 years, multifocal or bilateral PTC, organ-exceeding tumor growth (T4), metastasis (N1, M1), and familial accumulation of adolescent-onset thyroid disease. A hereditary predisposition to PTC is considered if both primary criteria or one primary criterion plus three secondary criteria are present. Family history-taking is recommended for all PTC patients to identify FPTC kindreds at risk. Blood relatives of FPTC index patients who harbor MNG should undergo thorough and regular clinical screening. Suspicious lesions should prompt early surgical intervention.