Progress in the Study of Gene Mutations Associated with Papillary Thyroid Carcinoma

In recent years, there has been a global rise in cases of papillary thyroid carcinoma (PTC), the predominant form of thyroid cancer. Advances in molecular biology have intensified the focus on the genetic mutations associated with this malignancy. Researchers have conducted extensive investigations into these mutations to elucidate their roles in the initiation, progression, treatment, and prognosis of PTC. This review synthesizes studies on the genetic mutations implicated in PTC, examining specific mutated genes, mechanisms of mutation, correlations with clinicopathological features, and their influence on treatment outcomes and prognosis. The objective is to provide a theoretical framework for enhancing the diagnosis, treatment, and prognostic assessment of PTC in the future.

Gene Expression of Fas,Soluble Fas and Fas-Ligand in Thyroid Tissues and Thyrocytes from Patients with Graves′ Disease

ObjectiveTo investigate Fas,soluble Fas(sFas)and Fas ligand(Fas L)gene expression in thyroid tissues and thyrocytes from patients with Graves disease(GD)and to find the interrelationship between apoptosis and pathogenesis of GD. MethodsThyroid tissues were obtained from 7 GD patients and 3 healthy subjects who died accidentally. Thyrocytes were cultured in Eagle′s medium. Total RNA was isolated from thyroid tissues and cultured thyrocytes. The cDNA was prepared by reverse transcription and amplified for Fas,sFas and Fas L by polymerase chain reaction(PCR). ResultsFas and sFas mRNA were detected in all samples from both GD and normal thyroid tissues and thyrocytes,but Fas L mRNA was only found in GD thyroid tissues and thyrocytes. Semi quantitative analysis showed that when compared with those of normal controls,the Fas and sFas mRNA levels were markedly increased in GD thyroid tissues(P<0.01),whereas in GD thyrocytes only the sFas mRNA levels was significantly elevated(P<0.01). ConclusionGene expression of Fas,sFas and Fas L showed abnormality in both thyroid tissues and thyrocytes from GD. The increased production of sFas might be involved in the hyperplasia of thyroid gland.

TSH RECEPTOR GENETIC ALTERATIONS IN THE AUTONOMOUSLY FUNCTIONING THYROID ADENOMAS

Objective To determine the relationship between TSH receptor gene mutations and autonomously functioning thyroid adenomas (AFTAs). Methods The thyroid samples from 14 cases of diagnosed AFTAs were analyzed, with normal thyroid specimens adjacent to the tumors as controls. The 155 base pairs DNA fragments which encompassed the third cytoplasmic loop and the sixth transmembrane segments in the TSH receptor gene exon 10 were amplified by Polymerase chain reaction (PCR) and analyzed by the single-strand conformation polymorphism (SSCP). Direct sequencing of the PCR products was performed with Prism Dye Terminator Cycle Sequencing Core Kit. Results 6 of 14 AFTA specimens displayed abnormal migration in SSCP analysis. In sequence analysis of 3 abnormally migrated samples, one base substitution at nucleotide 1957 (A to C) and two same insertion mutations of one adenosine nucleotide between nucleotide 1972 and 1973 were identified. No mutations were found in controls. Conclusion This study confirmed the presence of TSH receptor gene mutations in AFTAs; both one-point substitution mutation and one-base insertion mutation were found to be responsible for the pathogenesis of AFTAs.

Genetic Background May Confer Susceptibility to PTC in Benign Multinodular Thyroid Disease

Purpose: The incidence of hyperplastic thyroid nodular disease has been consistently rising over the last decades. In addition, unsuspected papillary thyroid carcinoma (PTC) can be found in up to 34% of patients operated for benign thyroid lesions. PTC tends to occur multi-focally and is commonly of polyclonal origin. We set out to test the hypothesis that in benign thyroid disease, a unique genetic signature can already be identified in the benign pathology, which is associated with a susceptibility of the thyroid tissue to neoplastic transformation in the context of additional growth promoting stimuli. Patients and Methods: We obtained a set of 23 samples from patients with multinodular goiter (MNG), 12 of whom also harbored an unsuspected PTC. We used global gene expression analysis to evaluate for dissimilarities in the gene expression patterns between these two groups. We also compared these patterns to the profiles of 3 normal thyroid and 7 PTC samples. Results: We were able to accurately distinguish between hyperplastic nodules of patients with multinodular goiter and those that were associated with a PTC. One of the strongest differentially expressed genes, CDC42, has been implicated to respond to environmental factors such as UVB radiation and might point to novel factors contributing to PTC genesis in the setting of pre-existing benign proliferative disease. Conclusion: While the comparison between histologically identical samples cannot distinguish the two groups of goiters, unsupervised or supervised approaches allowed us to identify a molecular signature associated with PTC susceptibility in multinodular goiter.

Autoimmune Thyroid Disease Genes Identified in Non-Caucasians

Autoimmune thyroid diseases (AITDs), including Graves’ disease (GD) and Hashimoto’s thyroiditis (HT), are among the commonest autoimmune disorders, affecting approximately 2% - 5% of the population. Epidemiological data support strong genetic influences on the development of AITD. The identification of genes placing individuals at an increased risk for the development of AITD has been a slow process. However, over the last 20 years or so real progress has been made with the mapping of novel loci, via a number of different approaches. The first AITD gene discovered, Human Leucocyte Antigen (HLA)/Major Histocompatibility Complex (MHC), is associated with both GD and HT. Non-MHC genes that confer susceptibility to AITD can be classified into two groups: (1) immune-regulatory genes (e.g., CD40, CTLA-4, and PTPN22);(2) thyroid-specific genes—thyroglobulin and TSH receptor genes. These genes interact with environmental factors, such as infection, likely through epigenetic mechanisms to trigger disease. In this review, we will summarize the latest findings on AITD susceptibility genes in non-Caucasians.

Identifying Driver Genes Mutations with Clinical Significance in Thyroid Cancer

Advances in technology are enabling gene mutations in papillary thyroid carcinoma(PTC)to be analyzed and clinical outcomes,such as recurrence,to be predicted.To date,the most common genetic mutation in PTC is in BRAF kinase(BRAF).However,whether mutations in other genes coincide with those in BRAF remains to be clarified.The aim of this study was to find mutations in other genes that co-exist with mutated BRAF,and to analyze their frequency and clinical relevance in PTC.Clinical and genetic data were collected from 213 PTC patients with a total of 36,572 mutation sites in 735 genes.After matching with genes from PTC entries in a global database(NCBI Gene),69 genes with mutations in coding regions were chosen for further study.Through frequency-based analysis,we identified commonly mutated genes co-existing with mutated BRAF and,using the mutation count correlation matrix(MCCM)method,analyzed their incidence according to age and gender.We designed Chord diagrams to reveal gene relationships concerning age and gender,and found that mutations in ALK,ATM,COL1A1,MSTIR,PRKCA,and WNK1 most commonly coincide with mutated BRAF,followed by APC,AURKA,and AURKB.These findings provide further insight into the genetic profile of PTC.

Screening and bioinformatics analysis of thyroid cancer-related hub genes

Objective:To identify the thyroid cancer-related hub genes and pathways by bioinformatics initially in order to lay the foundation for further study.Methods:The expression profile chips and data of thyroid cancer were screened and downloaded from the gene expression omnibus(GEO).The GEO2R was applied to identify the differential expressed genes between thyroid cancer tissues and normal thyroid tissues.And the Metascape online website was used for pathway and function enrichment.With the usage of STRING and Cytoscape,the protein-protein interaction network was constructed,and the plug-in app cytoHubba in Cytoscape was applied to screen hub genes.Kaplan-Meier Plotter was implemented to conduct survival analysis of hub genes for further screening and discussion.Results:A total of 304 differential expressed genes were screened,and were mainly enriched in the biological processes of extracellular matrix,cell-substrate adhesion,response to wounding,muscle structure development and hormone metabolic process etc.by Metascape.Protein-protein interaction network visualized 284 nodes;the top ten scores of Maximal Clique Centrality algorithm were taken as the criteria to screen out the hub genes with high connectivity in the gene expression network.The KM plotter analysis confirmed that 5 of 9 hub genes were correlated with the prognosis of thyroid cancer patients.Conclusion:FN1,SPP1,TIMP1,VCAN,COL1A1,COL1A2,MMP1,DCN,COMP and FMOD may play a significant role in the development of thyroid cancer.Genes which have prognostic significance in survival analyses were found to be relevant to the composition and regulation of extracellular matrix.

Evaluation of autophagy-related genes and lncRNAs signature for prognositic prediction in thyroid carcinoma via bioinformatics analysis

Autophagy plays a significant role in the pathogenesis and prognosis of thyroid carcinoma.The role of autophagy-related genes and long non-coding RNAs,as well as the risk model of thyroid carcinoma patients were investigated to predict clinical outcome of thyroid carcinoma.Different expression of autophagy-related genes and long non-coding RNAs in thyroid carcinoma patients was identified in The Cancer Genome Atlas database.Functional enrichment analysis and gene set enrichment analysis was used to hint the mechanism that autophagy might act in thyroid carcinoma.Univariate and multivariate Cox regression analyses were performed for screening the prognostic autophagy-related genes and long non-coding RNAs to construct prognostic related risk model.thyroid carcinoma patients were divided into the low-risk and high-risk groups.The overall survival time was both shorter in the high-risk groups than that in the low-risk groups.As for autophagy-related genes prognostic risk model,age and autophagy-related genes risk score are independent prognostic factors that affect the survival of thyroid carcinoma.ATIC and CDKN2A expression was closely related to pathological stage and T status,DNAJB1 expression was closely related to M status,age and gender.While autophagy-associated long non-coding RNA related prognostic risk model consequently demonstrated that the long non-coding RNA risk score could significantly predict the survival rate of thyroid carcinoma patients with areas under the curve of 0.972.gene set enrichment analysis presented that a total of 16 gene sets including 10 up-regulated and 6 down-regulated gene sets were significantly enriched.The autophagy-related genes and long non-coding RNAs based prognostic risk models are a reliable forecasting tool for thyroid carcinoma patients.

Research Progress on the Mechanism of Correlation Between Vitamin D and Thyroid Cancer

Vitamin D is a kind of fat-soluble vitamin,which is mainly involved in the metabolism of calcium and bone in the human body.As a metabolic substance,it also has a certain impact on the cellular microenvironment,and vitamin D also inhibits the proliferation of tumor cells.25(OH)D is considered the best index to evaluate the vitamin D level in the human body because of its relatively stable characteristics in the circulation.Thyroid cancer is a common malignant tumor that develops from malignant thyroid nodules.A large number of studies have found that the lower the serum 25(OH)D level,the higher the risk of thyroid nodules.A large number of studies have found that the lower the serum 25(OH)D level,the higher the risk of thyroid nodules.

甲状腺癌中NTRK基因融合的研究进展

神经营养性受体酪氨酸激酶(NTRK)基因融合是多种实体肿瘤的致癌驱动因素,是重要的临床生物标志物。在甲状腺癌中,NTRK基因融合阳性的患者表现出独特的临床病理特征,因此晚期、转移性或碘难治性甲状腺癌患者应该接受NTRK基因融合状态的评估。然而,关于甲状腺癌中NTRK基因融合的诊断和治疗目前还没有权威的建议或标准化的程序。本文综述了NTRK基因融合阳性甲状腺癌的研究进展,重点阐述了该疾病的临床病理特征,以及NTRK基因融合检测和靶向治疗药物的研究现状。

细针穿刺细胞学联合多基因检测在甲状腺结节诊断中的应用

目的探讨细针穿刺细胞学(FNAC)联合多基因检测在甲状腺结节诊断中的应用价值。方法选取符合纳入标准的204例甲状腺结节患者,所有患者均于术前行FNAC检查及多基因检测[BRAF、RAS(KRAS、HRAS、NRAS)、TERT、NTRK、RET/PTC共7个基因位点],以术后病理切片为“金标准”,分析FNAC联合多基因检测在甲状腺结节诊断中的价值。结果术后病理显示,FNAC联合多基因检测结果与病理结果一致性较强,Kappa值0.907,与FNAC和多基因突变单独检测相比,联合诊断的灵敏度更高,为97.54%,差异具有统计学意义(P<0.01),联合检测的准确率为95.59%,均高于FNAC和多基因突变单独检测,差异具有统计学意义(P>0.05)。结论因此,FNAC联合多基因检测可显著提高甲状腺结节诊断的准确率及灵敏度。

Thyroid hormone regulation of apoptotic tissue remodeling during anuran metamorphosis

Anuran metamorphosis involves systematic transformations of individual organs in a thyroid hormone (TH)-dependent manner. Morphological and cellular studies have shown that the removal of larval or- gans/tissues such the tail and the tadpole intestinal epithelium is through programmed cell death or apop- tosis. Recent molecular investigations suggest that TH regulates metamorphosis by regulating target gene expression through thyroid hormone receptors (TRs), which are DNA-binding transcription factors. Cloning and characterization of TH response genes show that diverse groups of early response genes are induced by TH. The products of these TH response genes are believed to directly or indirectly affect the expression and/or functions of cell death genes, which are conserved at both sequence and function levels in different animal species. A major challenge for future research lies at determining the signaling pathways leading to the activation of apoptotic processes and whether different death genes are involved in the regulation of apoptosis in different tissues/organs to effect tissue-specific transformations.

Involvement of chromatin and histone acetylation in the regulation of HIV-LTR by thyroid hormone receptor

The HIV-1 LTR controls the expression of HIV-1 viral genes and thus is critical for viral propagation and pathology. Numerous host factors have been shown to participate in the regulation of the LTR promoter. Among them is the thyroid hormone (T3) receptor (TR). TR has been shown to bind to the critical region of the promoter that contain the NFbB and Sp1 binding sites. Interestingly, earlier transient transfection studies in tissue culture cells have yielded contradicting conclusions on the role of TR in LTR regulation, likely due to the use of different cell types and/or lack of proper chromatin organization. Here, using the frog oocyte as a model system that allows replication-coupled chromatin assembly, mimicking that in somatic cells, we demonstrate that unliganded heterodimers of TR and RXR (9-cis retinoic acid receptor) repress LTR while the addition of T3 relieves the repression and further activates the promoter. More importantly, we show that chromatin and unliganded TR/RXR synergize to repress the promoter in a histone deacetylase-dependent manner.

Needle tract seeding of papillary thyroid carcinoma after fine-needle capillary biopsy:A case report

BACKGROUND Fine-needle biopsy is an accurate and cost-efficient tool for the assessment of thyroid nodules.It includes two primary methods:Fine-needle capillary biopsy(FNCB)and fine-needle aspiration biopsy.Needle tract seeding(NTS)is a rare complication of thyroid fine-needle biopsy mainly caused by fine-needle aspiration biopsy rather than FNCB.Here,we present an extremely rare case of a papillary thyroid carcinoma(PTC)patient with FNCB-derived NTS.CASE SUMMARY We report a 32-year-old woman with PTC who showed subcutaneous NTS 1 year after FNCB and thyroidectomy.NTS was diagnosed based on clinical manifestations,biochemistry indices,and imaging(computed tomography and ultrasound).Pathological identification of PTC metastases consistent with the puncture path is the gold standard for diagnosis.Surgical resection was the main method used to treat the disease.After surgery,thyroid function tests and ultrasound scans were performed every 3-6 mo.To date,no evidence of tumor recurrence has been observed.CONCLUSION FNCB is a safe procedure as NTS is rare,and can be easily removed surgically with no recurrence.Accordingly,NTS should not limit the usefulness of FNCB.

RAS AND p53 EXPRESSION IN HUMAN THYROID CARCINOMA

Objective: To investigate the possible interaction between the ras and p53 genes over-expression in thyroid carcinoma, and whether there is a correlation between the ras and p53 over-expression and clinicopathological criteria. Methods: Eighty patients with thyroid lesions were examined for expression of ras and p53 genes by the labeled streptavidin biotin peroxidase (LSAB) method. Of these patients, 54 were diagnosed (average age: 39.9±15.9 years) with malignant lesions. Of those included in the study, 31 has papillary carcinoma, 13 had follicular carcinoma, 7 had medullary carcinoma, 3 had undifferentiated carcinoma and 19 were stratified to stage I, 28 to stage II, 2 to stage III and 5 to stage IV according to TNM staging system. Twenty-six benign nodular thyroid disorders were studied as control. Results: Positive immunostain results for ras and p53 genes were statistically significant between thyroid carcinomas and benign disorders (90.7% vs 23%, 55.5% vs 30.7%,P<0.05). Both p53 and ras overexpressions coexisted in 30 thyroid carcinomas, and of these, 3 died and 5 had recurrences within 4 years. Conclusions: Activation of ras gene and inactivation of p53 gene were cooperatively associated in thyroid tumorigenesis. The concurrent overexpressions of ras and p53 could result in a poor prognosis.

Value of serum galectin-3 and midkine level determination for assessing tumor severity in patients with thyroid cancer

Objective:To investigate the value of serum galectin-3 and midkine level determination for assessing tumor severity in patients with thyroid cancer. Methods:108 patients with thyroid cancer treated in our hospital between January 2013 and February 2016 were collected and divided into I–II stage thyroid cancer group (n=39), III stage thyroid cancer group (n=55) and IV stage thyroid cancer group (n=14) according to TNM stage;56 patients diagnosed with thyroid adenoma after physical examination in our hospital during the same period were selected as control group. Enzyme-linked immunosorbent assay (ELISA) was used to determine serum galectin-3 and midkine levels of all groups;fluorescence quantitative PCR was used to determine proliferation gene and invasion gene mRNA expression in tumor tissue;Pearson test was used to analyze the correlation of serum galectin-3 and midkine levels with proliferation and invasion gene mRNA expression in patients with thyroid cancer. Results:Serum galectin-3 and midkine levels of patients with thyroid cancer were higher than those of control group (P<0.05), and the higher the tumor stage, the higher the serum galectin-3 and midkine levels;proliferation genes CyclinD1, BCL-2 and survivin mRNA expression in tumor tissue of patients with thyroid cancer were higher than those of control group (P<0.05) while p27 mRNA expression was lower than that of control group (P<0.05), invasion gene E-cadherin mRNA expression was lower than that of control group (P<0.05) while S100A4 and SATB1 mRNA expression were higher than those of control group (P<0.05), and the higher the tumor stage, the more significant the change of the above gene mRNA expression;Pearson test showed that serum galectin-3 and midkine levels in patients with thyroid cancer were correlated with proliferation and invasion gene expression in tumor tissue. Conclusions:Serum galectin-3 and midkine levels can directly reflect the malignant degree of thyroid cancer, and is expected to become the new means for early disease diagnosis, curative effect evaluation and prognosis judgment.

Thyroid Disrupting Activities in Wastewater and Surface Waters: A Review of Their <i>In Vitro</i>Analyses

The developmental and reproductive effects of endocrine disruption on humans and wildlife rank among the most threatening of all environmental health concerns. Particularly vulnerable to chemical assaults are the hypothalamic-pituitary-gonadal (HPG) axis and the hypothalamic-pituitary-thyroid (HPT) axis of the endocrine system. While the effects of endocrine disrupting chemicals (EDCs) on the HPG axis have been the subject of intense research efforts, with comprehensive elucidation, a lot remains to be clarified on the effects of EDCs on thyroid functions. For instance, there are no clear-cut biomarkers of exposure and effects of thyroid disrupting chemicals (TDCs) in intact organisms. Consequently, a number of in vitro assays have been developed, and are particularly useful for the identification and mechanistic characterization of potential TDCs considering the increasing number of EDCs that are being released into the environment. However, with the in vitro assays, studies suggest that a plausible major mode of action of TDCs, thyroid hormone receptor (THR) agonist activity, is not environmentally relevant. Here, we reviewed in vitro detection of TDCs activities in wastewater and surface waters. Data strongly suggest that cell lines may be responsible for the less frequent detection of THR agonist activity in wastewater and surface water. It was concluded that the development of reporter gene assays with thyroid hormone function related cell lines, is required.

THE EFFECT OF FCγ RECEPTOR ON THE PATHOGENESIS OF GRAVES' DISEASE

Objective To explore the roles of Fcγ recep tor in the pathogenesis of Graves' disease. Methods Fcγ receptor gene knockout mice(Fcγ R KO m ice) which were rooted in C57BL/6 mice and wild type C57BL/6 mice were immunized by hTSH receptor expressing cells (DAP3.WT). 1-2×107 DAP3.WT cells were peri toneally injected into mice every two weeks for a total of six times. Two weeks after final immunization, mice were killed for measurement of total thyroxine, T RAb and pathological examination. Results The thyroxine level of the immunized Fcγ recept or gene knockout mice was significantly lower than that of the immunized wild ty pe control mice (2.2±0.31 vs. 3.32± 0.59 g·dL -1, P< 0.05 ),but there was no significant difference between immunized Fcγ R KO mic e and non-immunized wild type control group. The TRAb levels of the immunized F γ R KO mice significantly increased compared to those of the immunized wild type mice (21.75±8.21 vs. 14.11±6.21, P< 0.05). The lymphocyte cel ls infiltration and destruction of thyroid follicles were found in the thyroid gland of the immunized Fcγ R KO mice. Conclusion These results suggest that Fcγ receptor may be involved in the pathogenesis of Graves' disease.

TMEM158 May Serve as a Diagnostic Biomarker for Anaplastic Thyroid Carcinoma:An Integrated Bioinformatic Analysis

Anaplastic thyroid carcinoma(ATC)is a rare but extremely lethal malignancy.However,little is known about the pathogenesis of ATC.Given its high mortality,it is critical to improve our understanding of ATC pathogenesis and to find new diagnostic biomarkers.In the present study,two gene microarray profiles(GSE53072 and GSE65144),which included 17 ATC and 17 adjacent non-tumorous tissues,were obtained.Bioinformatic analyses were then performed.Immunohistochemistry(IHC)and receiver operating characteristic(ROC)curves were then used to detect transmembrane protein 158(TMEM158)expression and to assess diagnostic sensitivity.A total of 372 differentially expressed genes(DEGs)were identified.Through protein-protein interaction(PPI)analysis,we identified a significant module with 37 upregulated genes.Most of the genes in this module were related to cell-cycle processes.After co-expression analysis,132 hub genes were selected for further study.Nine genes were identified as both DEGs and genes of interest in the weighted gene co-expression network analysis(WGCNA).IHC and ROC curves confirmed that TMEM158 was overexpressed in ATC tissue as compared with other types of thyroid cancer and normal tissue samples.We identified 8 KEGG pathways that were associated with high expression of TMEM158,including aminoacyl-tRNA biosynthesis and DNA replication.Our results suggest that TMEM158 may be a potential oncogene and serve as a diagnostic indicator for ATC.

The correlation between the ultrasonic elastic strain ratio of thyroid cancer and the malignant biological characteristics of cancer cells in the lesions

Objective:To study the correlation between the ultrasonic elastic strain ratio of thyroid cancer and the malignant biological characteristics of cancer cells in the lesions.Methods:A total of 90 patients with thyroid cancer who accepted surgical treatment in our hospital between March 2015 and September 2016 were selected as the observation group, and 50 patients who received surgery in our hospital during the same period and were with clear pathological diagnosis of thyroid adenoma were selected as the control group. According to the median of ultrasonic elastic strain ratio, the observing group were further divided into high elastic strain ratio group and low elastic strain ratio group (n=45). RT-PCR was used to determine the mRNA expression of proliferation genes, invasion genes and autophagy genes in lesions of all groups of patients.Results: The mean elastic strain ratio of observation group was significantly lower than that of control group. Proliferation gene FOXA1 mRNA expression in lesions of low elastic strain ratio group and high elastic strain ratio group were higher than those of control group while PP4R1 and Smac mRNA expression were lower than those of control group, and as the elastic strain ratio decreased, the FOXA1 mRNA expression increased while PP4R1 and Smac mRNA expression decreased;invasion genes BCORL1, SATB1, Twist1 and CXCR4 mRNA expression in lesions of low elastic strain ratio group and high elastic strain ratio group were higher than those of control group, and as the elastic strain ratio decreased, the BCORL1, SATB1, Twist1 and CXCR4 mRNA expression increased;autophagy genes ARHI, Beclin1, LC3 and PTEN mRNA expression in lesions of low elastic strain ratio group and high elastic strain ratio group were lower than those of control group, and as the elastic strain ratio decreased, the ARHI, Beclin1, LC3 and PTEN mRNA expression decreased.Conclusion:The ultrasonic elastic strain ratio of thyroid cancer is negatively correlated with tumor cell proliferation and invasion activity, and positively correlated with autophagy activity.