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

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
S. Adeleh Razavi ◽  
Mandana Afsharpad ◽  
Mohammad Hossein Modarressi ◽  
Maryam Zarkesh ◽  
Parichehreh Yaghmaei ◽  
...  
Keyword(s):  
Papillary Thyroid Carcinoma ◽  
Thyroid Carcinoma ◽  
Internal Control ◽  
Reference Genes ◽  
Thyroid Tissue ◽  
Optimal Number ◽  
Thyroid Tumors ◽  
Papillary Thyroid ◽  
Tissue Samples ◽  
Qrt Pcr

Abstract 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.

Identification of Differentially Expressed Genes Associated with Papillary Thyroid Carcinoma

2020 ◽  
Vol 23 (6) ◽  
pp. 546-553
Author(s):  
Hongyuan Cui ◽  
Mingwei Zhu ◽  
Junhua Zhang ◽  
Wenqin Li ◽  
Lihui Zou ◽  
...  
Keyword(s):  
Papillary Thyroid Carcinoma ◽  
Thyroid Carcinoma ◽  
Differentially Expressed Genes ◽  
Molecular Mechanisms ◽  
Cellular Proliferation ◽  
Mrna Level ◽  
Thyroid Tissue ◽  
Differentially Expressed ◽  
Thyroid Tumors ◽  
Papillary Thyroid

Objective: Next-generation sequencing (NGS) was performed to identify genes that were differentially expressed between normal thyroid tissue and papillary thyroid carcinoma (PTC). Materials & Methods: Six candidate genes were selected and further confirmed with quantitative real-time polymerase chain reaction (qRT-PCR), and immunohistochemistry in samples from 24 fresh thyroid tumors and adjacent normal tissues. The Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis was used to investigate signal transduction pathways of the differentially expressed genes. Results: In total, 1690 genes were differentially expressed between samples from patients with PTC and the adjacent normal tissue. Among these, SFRP4, ZNF90, and DCN were the top three upregulated genes, whereas KIRREL3, TRIM36, and GABBR2 were downregulated with the smallest p values. Several pathways were associated with the differentially expressed genes and involved in cellular proliferation, cell migration, and endocrine system tumor progression, which may contribute to the pathogenesis of PTC. Upregulation of SFRP4, ZNF90, and DCN at the mRNA level was further validated with RT-PCR, and DCN expression was further confirmed with immunostaining of PTC samples. Conclusion: These results provide new insights into the molecular mechanisms of PTC. Identification of differentially expressed genes should not only improve the tumor signature for thyroid tumors as a diagnostic biomarker but also reveal potential targets for thyroid tumor treatment.

scholarly journals Single-cell RNA sequencing demonstrates the intratumoral heterogeneityof papillary thyroid carcinoma

2021 ◽  
Author(s):  
Zhengshi Wang ◽  
Youlutuziayi Rixiati ◽  
Wenli Jiang ◽  
Caiguo Huang ◽  
Binghua Jiao ◽  
...  
Keyword(s):  
Papillary Thyroid Carcinoma ◽  
Thyroid Carcinoma ◽  
Single Cell ◽  
Rna Sequencing ◽  
Immune Therapy ◽  
Thyroid Tissue ◽  
Papillary Thyroid ◽  
Sequencing Data ◽  
Tissue Samples ◽  
Single Cell Rna Sequencing

AbstractPapillary thyroid carcinoma(PTC) is the most common thyroid malignancy, to investigate the intratumoral heterogeneity of PTC, we analyzed single-cell RNA-sequencing data and identified 10 major cell types from primary papillary thyroid carcinoma, lymph metastatic, or paired normal thyroid tissue samples. In this study, we verified that the increase in the proportion of CD4+Tregs may be key factor responsible for the immunosuppressive property of PTC. Inhibitory checkpoints, such as TIGIT and CD96 may be better targets for immune therapy in lymph metastatic papillary thyroid carcinoma. Our results will further the understanding of the heterogeneity among papillary thyroid carcinoma and provide an essential resource for drug discovery in the future.

scholarly journals Genetic Determinants for Prediction of Outcome of Patients with Papillary Thyroid Carcinoma

Cancers ◽  
2021 ◽  
Vol 13 (9) ◽  
pp. 2048
Author(s):  
Antónia Afonso Póvoa ◽  
Elisabete Teixeira ◽  
Maria Rosa Bella-Cueto ◽  
Rui Batista ◽  
Ana Pestana ◽  
...  
Keyword(s):  
Papillary Thyroid Carcinoma ◽  
Thyroid Carcinoma ◽  
Patient Outcomes ◽  
Genetic Alterations ◽  
Papillary Thyroid ◽  
Tissue Samples ◽  
Primary Tumors ◽  
Persistent Disease ◽  
Increased Risk ◽  
Structural Disease

Papillary thyroid carcinoma (PTC) usually presents an excellent prognosis, but some patients present with aggressive metastatic disease. BRAF, RAS, and TERT promoter (TERTp) genes are altered in PTC, and their impact on patient outcomes remains controversial. We aimed to determine the role of genetic alterations in PTC patient outcomes (recurrent/persistent disease, structural disease, and disease-specific mortality (DSM)). The series included 241 PTC patients submitted to surgery, between 2002–2015, in a single hospital. DNA was extracted from tissue samples of 287 lesions (primary tumors and metastases). Molecular alterations were detected by Sanger sequencing. Primary tumors presented 143 BRAF, 16 TERTp, and 13 RAS mutations. Isolated TERTpmut showed increased risk of structural disease (HR = 7.0, p < 0.001) and DSM (HR = 10.1, p = 0.001). Combined genotypes, BRAFwt/TERTpmut (HR = 6.8, p = 0.003), BRAFmut/TERTpmut (HR = 3.2, p = 0.056) and BRAFmut/TERTpwt (HR = 2.2, p = 0.023) showed increased risk of recurrent/persistent disease. Patients with tumors BRAFwt/TERTpmut (HR = 24.2, p < 0.001) and BRAFmut/TERTpmut (HR = 11.5, p = 0.002) showed increased risk of structural disease. DSM was significantly increased in patients with TERTpmut regardless of BRAF status (BRAFmut/TERTpmut, log-rank p < 0.001; BRAFwt/TERTpmut, log-rank p < 0.001). Our results indicate that molecular markers may have a role in predicting PTC patients’ outcome. BRAFmut/TERTpwt tumors were prone to associate with local aggressiveness (recurrent/persistent disease), whereas TERTpmut tumors were predisposed to recurrent structural disease and DSM.

scholarly journals Break–apart interphase fluorescence in situ hybridization assay in papillary thyroid carcinoma: on the road to optimizing the cut-off level for RET/PTC rearrangements

2015 ◽  
Vol 172 (5) ◽  
pp. 571-582 ◽  
Author(s):  
Chiara Colato ◽  
Caterina Vicentini ◽  
Silvia Cantara ◽  
Serena Pedron ◽  
Paolo Brazzarola ◽  
...  
Keyword(s):  
Papillary Thyroid Carcinoma ◽  
Thyroid Carcinoma ◽  
Chromosomal Rearrangements ◽  
Study Cohort ◽  
Fish Analysis ◽  
Papillary Thyroid ◽  
Qrt Pcr ◽  
Molecular Events ◽  
On The Road

ObjectiveChromosomal rearrangements of theRETproto-oncogene is one of the most common molecular events in papillary thyroid carcinoma (PTC). However, their pathogenic role and clinical significance are still debated. This study aimed to investigate the prevalence of RET/PTC rearrangement in a cohort ofBRAFWT PTCs by fluorescencein situhybridization (FISH) and to search a reliable cut-off level in order to distinguish clonal or non-clonal RET changes.DesignFortyBRAFWT PTCs were analyzed by FISH for RET rearrangements. As controls, sixBRAFV600E mutated PTCs, 13 follicular adenomas (FA), and ten normal thyroid parenchyma were also analyzed.MethodsWe performed FISH analysis on formalin-fixed, paraffin-embedded tissue using a commercially available RET break–apart probe. A cut-off level equivalent to 10.2% of aberrant cells was accepted as significant. To validate FISH results, we analyzed the study cohort by qRT-PCR.ResultsSplit RET signals above the cut-off level were observed in 25% (10/40) of PTCs, harboring a percentage of positive cells ranging from 12 to 50%, and in one spontaneous FA (1/13, 7.7%). Overall, the data obtained by FISH matched well with qRT-PCR results. Challenging findings were observed in five cases showing a frequency of rearrangement very close to the cut-off.ConclusionsFISH approach represents a powerful tool to estimate the ratio between broken and non-broken RET tumor cells. Establishing a precise FISH cut-off may be useful in the interpretation of the presence of RET rearrangement, primarily when this strategy is used for cytological evaluation or for targeted therapy.

Papillary Thyroid Carcinoma in Ectopic Thyroid Tissue Within a Suspected Cervical Paraganglioma

2021 ◽  
Vol Publish Ahead of Print ◽  
Author(s):  
Marcel Ernesto Sambo Salas ◽  
Diego Muñoz Moreno ◽  
Laura Gonzalez Fernandez ◽  
Patricia Nuñez Ramos ◽  
Maria Cebollero Presmanes
Keyword(s):  
Papillary Thyroid Carcinoma ◽  
Thyroid Carcinoma ◽  
Thyroid Tissue ◽  
Ectopic Thyroid ◽  
Papillary Thyroid ◽  
Ectopic Thyroid Tissue

Cadmium as main endocrine disruptor in papillary thyroid carcinoma and the significance of Cd/Se ratio for thyroid tissue pathophysiology

2019 ◽  
Vol 55 ◽  
pp. 190-195 ◽  
Author(s):  
Aleksandar Stojsavljević ◽  
Branislav Rovčanin ◽  
Đurđa Krstić ◽  
Jovana Jagodić ◽  
Slavica Borković-Mitić ◽  
...  
Keyword(s):  
Papillary Thyroid Carcinoma ◽  
Thyroid Carcinoma ◽  
Endocrine Disruptor ◽  
Thyroid Tissue ◽  
Papillary Thyroid

scholarly journals Pre-sternal thyroid swellings: a case of rare aberrant site recurrence and review of literature

2019 ◽  
Vol 12 (1) ◽  
Author(s):  
Lovenish Bains ◽  
Sushant Bhatia ◽  
Rohit Kaushik ◽  
Sudhir Kumar Jain ◽  
Chandra Bhushan Singh ◽  
...  
Keyword(s):  
Lymph Node ◽  
Papillary Thyroid Carcinoma ◽  
Thyroid Carcinoma ◽  
De Novo ◽  
Anterior Part ◽  
Thyroid Tissue ◽  
Papillary Thyroid ◽  
Thyroid Cells ◽  
Thyroid Swelling ◽  
The Right

Abstract Background Thyroid swellings enlarge caudally into the mediastinum behind the sternum. Pre-sternal swelling of thyroid origin is very rare. We present our case of pre-sternal thyroid swelling which was albeit a surprisingly rare site of papillary thyroid carcinoma recurrence and review of pre-sternal thyroid swellings reported till date. Case summary A 60 year old female presented with a painless, progressive swelling on the anterior part of the chest for the past 2 years. A 15 cm × 8 cm vertically aligned, non tender, well defined swelling was present on the pre-sternal region, with consistency ranging from soft to firm. The swelling was fixed to the underlying tissues and a fixed level IV lymph node was palpable on the right side. Ultrasonography revealed a large mass of 15 × 7 cm with multiple cystic areas. Fine needle aspiration cytology was inconclusive twice. Patient had undergone a total thyroidectomy for papillary carcinoma 10 years back. Computed tomography findings revealed a large 15 × 6.6 × 7 cm lobulated, pre-sternal, soft tissue lesion with solid & cystic components. The mass was infiltrating the right sided strap muscles and sternocleidomastoid. FNAC was inconclusive and thyroid scan could not pick up any activity in the mass. Henceforth a PET scan was done that showed increased FDG uptake by the lesion and the level IV lymph node. The patient underwent wide excision of the mass with right functional neck dissection, along with removal with both sternal head of sternocleido-mastoid, the strap muscles and the surrounding fascia. Histopathology confirmed papillary thyroid carcinoma. Patient received post-operative radioactive iodine ablation and is healthy with no recurrence up to 30 months of follow up. Discussion The mechanisms for pre-sternal thyroid swelling are not understood due to paucity of cases. The mechanisms proposed are invasion of strap muscles and cervical linea alba and tumor cells spread anterior to sternum, truly ectopic thyroid tissue, de novo carcinogenesis in the embryonal remnants like the thyro-thymic residues, sequestered thyroid tissue which grows later or migration of thyroid cells, incomplete clearance at the time of primary surgery or intraoperative seeding. Conclusion Pre-sternal region masses of thyroid origin are very rare. A proper work up, suspicion for thyroid mass and array of tests will be required to come to a provisional diagnosis. Since the masses reported in literature were primarily malignant, any such mass may be treated on lines of malignancy with radical surgery.

scholarly journals Occult Papillary Thyroid Carcinoma without Detection of the Primary Tumor on Preoperative Ultrasonography or Postoperative Pathological Examination: A Case Report

2020 ◽  
Vol 13 (1) ◽  
pp. 105-112 ◽  
Author(s):  
Gai Yamashita ◽  
Takahito Kondo ◽  
Akira Okimura ◽  
Munehide Nakatsugawa ◽  
Hiroshi Hirano ◽  
...  
Keyword(s):  
Lymph Node ◽  
Papillary Thyroid Carcinoma ◽  
Thyroid Carcinoma ◽  
Primary Tumor ◽  
Thyroid Tissue ◽  
Pathological Examination ◽  
Papillary Thyroid ◽  
Superficial Lobe ◽  
Preoperative Ultrasonography ◽  
The Right

Herein, we report a case of an occult thyroid cancer that was not detected as a primary tumor on preoperative ultrasonography or postoperative pathological examination, although a diagnosis of papillary thyroid carcinoma metastasis was made owing to the presence of a mass in the right upper neck. Needle biopsy of the mass in the right upper neck revealed positive results for thyroglobulin and TTF-1 on immunostaining, and a papillary thyroid carcinoma was observed with papillary and follicular patterns. We suspected papillary thyroid carcinoma (T0N1bM0) or ectopic papillary thyroid carcinoma. Accordingly, we performed total thyroidectomy, central lymph node dissection, right lateral neck dissection, and resection of the superficial lobe of the right parotid. A postoperative pathological examination of 5-mm slices of the specimen revealed no primary tumor in the thyroid. However, a hyalinized image of the thyroid indicated that a micropapillary thyroid carcinoma might have spontaneously disappeared. As there was no normal thyroid tissue in the metastasis to the superior internal jugular lymph node, the tumor was unlikely to be an ectopic papillary thyroid carcinoma. Therefore, we made a diagnosis of a papillary thyroid carcinoma (pT0N1bM0). After surgery, we determined that the tumor belonged to a high-risk group of papillary thyroid carcinomas and a poor-prognosis group of symptomatic papillary thyroid microcarcinomas; accordingly, ablation was performed with 30 mCi iodine-131. There was no recurrence or metastasis 24 months after the first surgery.

scholarly journals Clinicopathological Significance of Loss of p27Kip1 Expression in Papillary Thyroid Carcinoma

2017 ◽  
Vol 32 (2) ◽  
pp. 255-259 ◽  
Author(s):  
Nae Yu Kim ◽  
Jin Hwa Kim ◽  
Jung-Soo Pyo ◽  
Won Jin Cho
Keyword(s):  
Papillary Thyroid Carcinoma ◽  
Thyroid Carcinoma ◽  
Meta Analysis ◽  
Thyroid Tissue ◽  
Clinicopathological Characteristics ◽  
Papillary Thyroid ◽  
Benign Lesions ◽  
Thyroid Microcarcinoma ◽  
Clinicopathological Significance ◽  
Expression Loss

Introduction A meta-analysis was done to investigate the clinicopathological significance of the loss of p27kip1 expression in papillary thyroid carcinoma (PTC). Methods The meta-analysis involving 17 studies included 1,652 PTC and 328 benign cases. The rate of p27kip1 expression loss in PTC and benign lesions, and the correlations between p27kip1 expression loss and clinicopathological characteristics of PTC were determined. Results The estimated rate of p27kip1 expression loss was 0.557 (95% confidence interval [CI] 0.443-0.665) and 0.139 (95% CI 0.062-0.283) in PTC and benign lesions, respectively. In subgroup analysis, the rates of p27kip1 expression loss were 0.683, 0.393, and 0.414 in the classical variant, follicular variant, and papillary thyroid microcarcinoma, respectively. Loss of p27kip1 expression was significantly correlated with lymph node metastasis and distant metastasis (odds ratio 3.559, 95% CI 1.146-11.056 and 4.735, 95% CI 1.322-16.960, respectively). Extrathyroidal extension was correlated with loss of p27kip1 expression, but not in a statistically significant way (p = 0.051). There were no significant correlations between loss of p27kip1 expression and sex, tumor size, BRAFV600E mutation, and tumor multifocality. Conclusions Loss of p27kip1 expression is frequently found in PTC compared with benign lesions and normal thyroid tissue. When present in PTC, it is correlated with aggressive tumor behavior.

Papillary thyroid carcinoma in a lateral neck cyst: primary of ectopic thyroid tissue versus cystic metastasis

2013 ◽  
Vol 127 (7) ◽  
pp. 724-727 ◽  
Author(s):  
J J Xu ◽  
K Kwan ◽  
K Fung
Keyword(s):  
Papillary Thyroid Carcinoma ◽  
Thyroid Gland ◽  
Thyroid Carcinoma ◽  
Papillary Carcinoma ◽  
Thyroid Tissue ◽  
Ectopic Thyroid ◽  
Cystic Degeneration ◽  
Lateral Neck ◽  
Papillary Thyroid ◽  
Ectopic Thyroid Tissue

AbstractObjective:To review the diagnosis of primary papillary carcinoma of ectopic thyroid tissue within branchial cleft cysts, and to discuss the diagnostic challenge of differentiating this condition from metastatic disease when an occult microcarcinoma is found in the thyroid gland.Methods:These comprise a case report and a literature review. We present the case of a 75-year-old woman with papillary thyroid carcinoma within the wall of a recurrent, 15 cm, lateral neck cyst.Results:Histological examination of the patient's thyroid gland found a 0.5 mm papillary thyroid microcarcinoma.Conclusion:Our differential diagnosis was primary papillary carcinoma arising from ectopic thyroid tissue, or metastatic cystic degeneration of a lateral lymph node. We make an argument for the former.

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