Iodine nutritional status is not a direct factor in the prevalence of the BRAFV600E mutation in papillary thyroid cancer

ABSTRACT Objectives: Papillary thyroid carcinoma (PTC) accounts for approximately 85%-90% of all thyroid cancers. Of the iodine-metabolizing genes, BRAFV600E is a highly specific target for PTC and may have a reciprocal causative relationship with iodide-metabolizing genes. Materials and methods: In this study, we performed a data analysis of selected quantitative studies to determine the relationship between iodine nutritional status and the prevalence of the BRAF600E mutation in patients with PTC. Five studies were selected for meta-analysis based on the selection criteria. Results: A total of 2,068 patients were divided into three groups: low (urinary iodine concentration [UIC] < 100 μg/L), adequate (UIC 100-200 μg/L), and high (UIC ≥ 200 μg/L). The results were obtained using RevMan software, and the pooled odds ratios (ORs) were calculated using Mantel-Haenszel statistics with a 95% confidence interval (CI). The OR for the prevalence of the BRAFV600E mutation between the high and adequate groups was 1.25 (95% CI 0.64-2.43, p = 0.51), and the OR between the low and adequate groups was 0.98 (95% CI 0.42-2.31, p = 0.96). The BRAFV600E mutation risk did not change significantly at different levels of iodine nutrition (p = 0.33) in statistical analyses. Conclusion: We conducted preliminary research on dietary iodine intake and the BRAFV600E mutation in PTC. The results suggested that abnormal iodine intake might not directly influence the prevalence of the BRAFV600E mutation in these patients. Further research into the associations between dietary iodine intake and the BRAFV600E mutation in PTC, including the underlying mechanisms, is required.

O receptor aril de hidrocarbonetos (AhR) em melanomas: associação com mutações BRAF, com a progressão da doença e identificação de ligantes / The aryl hydrocarbon receptor (AhR) in melanomas: association with BRAF mutations, with disease progression and ligand identification

O melanoma é um tipo de câncer de pele geneticamente diverso, que surge diante das transformações em melanócitos. A mutação BRAFV600E está presente em mais de 90% de todas as mutações em BRAF, sendo assim ocorre em cerca de 50% dos casos registrados. As mutações em NRAS, ocupam o segundo lugar entre as mutações mais prevalentes, cerca de 20% dos casos. Informações sobre as assinaturas genéticas, permitiram o desenvolvimento de terapia alvo dirigida. O Vemurafenib, inibidor da quinase BRAFV600E, apresentou inicialmente resultados bastante satisfatórios, contudo existe registro de casos de recidiva e resistência. O receptor aril de hidrocarbonetos é expresso em vários componentes da pele, e assim está relacionado a homeostase e fisiopatologia da pele. Diante disso, a avaliação da expressão do receptor em um painel de linhagens mutadas para NRAS e BRAF, e BRAF resistentes, mostrou-se maior do que a encontrada em melanócitos. Também encontramos maior expressão de mRNA de AhR em linhagens de melanoma derivadas de sítio primário e metastático, mutadas para BRAFV600E, quando comparadas ao melanócito. Agregado a isto, a análise in silico no TCGA (The Cancer Genome Atlas) mostrou que há 18% de alteração genética em AhR, sendo em maior parte a alta regulação de mRNA. Também, a análise do banco público GSE12391, mostrou aumento de mRNA de AhR na fase de crescimento vertical do melanoma. Assim, concluímos que há maior expressão de mRNA e sua importância nas fases de desenvolvimento do melanoma, tanto nos processos iniciais quanto em processos de migração, invasão e metástase. Ainda, encontramos maior mRNA do receptor em linhagens resistentes ao Vemurafenib. Este resultado sustenta a hipótese de que AhR pode ser considerado um marcador de resistência em melanomas. O AhR, inicialmente no citoplasma, quando ativado pode atuar como fator de transcrição regulando vários genes que apresentam sequências definidas, participando de respostas carcinogênicas. Compostos halogenados e moléculas endógenas derivadas das vias de metabolização do triptofano são agonistas do receptor. Anteriormente, nosso grupo mostrou que linhagens de melanoma incubadas com triptamina e DMT exibiram menor clonogenicidade. Diante de uma literatura escassa sobre o papel do DMT no melanoma e com base nestes resultados, nosso objetivo foi avaliar o papel de AhR nesta interface DMT-melanoma. Para isto, nosso objetivo foi construir linhagem editada geneticamente para AhR através da ferramenta CRISPR-Cas9. Vários foram os esforços, sem sucesso, utilizados nas tentativas de comprovar a manutenção de células editadas na cultura. Atrelamos a este resultado a possibilidade de haver duas subpopulações editadas geneticamente pós CRISPR-Cas9, onde uma destas manteve o padrão de crescimento semelhante às células wild type. Devido a este crescimento diferencial, não obtivemos congruências nos ensaios e postulamos a perda do possível nocaute. A partir disso, realizamos ensaios de interactoma para avaliar a interação de DMT-AhR. Nosso resultado sugere a interação de DMT ao receptor sigma 1, e não ao receptor aril de hidrocarbonetos. Desta forma, o interactoma sustenta a hipótese de que DMT não é um ligante de AhR. Para certificar este resultado análises de docking associados a ensaios biológicos, avaliando o papel do receptor, devem ser realizados para averiguar a afinidade e seletividade de DMT como ligante do receptor na linhagem de melanoma

Melanoma is a genetically diverse type of skin cancer, which arises from changes in melanocytes. The BRAFV600E mutation is present in more than 90% of all BRAF mutations, so it occurs in about 50% of registered cases. Mutations in NRAS occupy the second place among the most prevalent mutations, about 20% of cases. Information on genetic signatures allowed the development of targeted therapy. vemurafenib, kinase inhibitor BRAFV600E, initially presented very satisfactory results, however there is a record of cases of relapse and resistance. The aryl hydrocarbon receptor is expressed in several components of the skin and is thus related to homeostasis and skin pathophysiology. Therefore, the evaluation of receptor expression in a panel of strains mutated to NRAS and BRAF, and resistant BRAF, proved to be greater than that found in melanocytes. We also found main expression of AhR mRNA in melanoma strains derived from primary and metastatic site, mutated to BRAFV600E, when compared to melanocyte. Added to this, the in silico analysis in TCGA (The Cancer Genome Atlas) showed that there is 18% of genetic alteration in AhR, being mostly the high regulation of mRNA. Also, an analysis by the public bank GSE12391, showed an increase in AhR mRNA in the vertical growth phase of melanoma. Thus, it is concluded that there is greater expression of mRNA and its importance in the stages of development of melanoma, both in recent processes and in the processes of migration, invasion and metastasis. In addition, we found higher receptor mRNA in strains resistant to vemurafenib. This result supports the hypothesis that AhR can be considered a marker of resistance in melanomas. AhR, initially in the cytoplasm, when activated can act as a transcription factor regulating several genes that have defined sequences, participating in carcinogenic responses. Along with this, we show that along the tumor progression, there is an increase in AhR in the radial growth phase of melanoma. Halogenated compounds and endogenous molecules derived from the tryptophan metabolism pathways are receptor agonists. Previously, our group showed that melanoma strains incubated with tryptamine and DMT exhibited less clonogenicity. In view of a scarce literature on the role of DMT in melanoma and based on these results, our objective was to evaluate the role of AhR in this DMT-melanoma interface. For this, our goal was to build genetically edited strain for AhR using the CRISPR-Cas9 tool. Several efforts were unsuccessful in attempts to prove the maintenance of cells edited in the culture. We linked to this result the possibility of having two subpopulations genetically edited after CRISPR-Cas9, where one of them maintained the growth pattern like wild type cells. Due to this differential growth, we did not obtain congruence in the tests and postulated the loss of the possible knockout. From that, we performed interactome assays to evaluate the DMT-AhR interaction. Our result suggests the interaction of DMT with the sigma 1 receptor, and not the aryl hydrocarbon receptor. Thus, the interactome supports the hypothesis that DMT is not an AhR ligand. To certify this result, docking analyses associated with biological assays, evaluating the role of the receptor, should be performed to ascertain the affinity and selectivity of DMT as a ligand of the receptor in the melanoma lineage

“Black Ink” Sonographic Pattern as a Predictor for Invasive Papillary Thyroid Micro Carcinoma: A Case Report

Background:Thisstudy investigated the utility of the Sonographic pattern "BlackInk" with BRAF mutation testing of thyroid fine-need leaspi rationcy to logy specimens for the risk papil larythy roidmic rocarcinoma(PTMC).Case Presentation: Wed escribea caseofa41-year-old Caucasian woman af fected by ault rason ography “BlackInk”papillarythyroidmicrocarcinoma(PTMC)oftheleftlobeofthethyroidglandwithverytinysize(Ø0.4cm).Thecharacteristics,withtheDiagnosticImagingusingUltrasonography(US),SuperbMicro-VascularImaging(SMI),fine-needle-aspirationcytology(FNAC)andmutationanalysisareherediscussed.Therearemoreraresubtypesofthyroidcanceraspapillarymicrocarcinoma"BlackInk"thatevenifsmall,areinvasiveandthereiswhytheneedtoearlydiagnosistoavoidtheiraggressivebehaviorisneeded.Nowadays,focusingonthesize,thecut-offfornon-occulttinytumorshasdroppedto0.3cm.Thisvalueisofgreatrelevance.Conclusion:Ultrasonography,FNACandBRAFmolecularstudyhaveproventobethemostsensitivediagnosticcombinationfortheearlydetectionofthyroidcancer.Despitethesizeofthismicro-lesion,theBlackInkultrasonographicpatternassociatedwithmalignantcytologyatFNACrepresentsanimportantbiologicalriskfactorandcouldstillbeapredictorofthePTMCandriskfornecklymphnodemetastases

Preoperative detection of TERT promoter and BRAFV600E mutations in papillary thyroid carcinoma in high-risk thyroid nodules

ABSTRACT Objectives: This observational study analyzed telomerase reverse transcriptase (pTERT) mutations in 45 fine-needle aspiration (FNA) specimens obtained from thyroid nodules followed by postoperatively confirmation of papillary thyroid cancer (PTC) diagnosis, examining their relationship with clinicopathologic aspects and the BRAFV600E mutation. Subjects and methods: Clinical information was collected from patients who presented to Ribeirao Preto University Hospital for surgical consultation regarding a thyroid nodule and who underwent molecular testing between January 2010 to October 2012. Tests included a DNA-based somatic detection of BRAFV600E and pTERT mutations. Results: We found coexistence of pTERTC228T and BRAFV600E mutations in 8.9% (4/45) of thyroid nodules. All nodules positive for pTERT mutations were BRAFV600E positives. There was a significant association between pTERTC228T/BRAFV600E with older age and advanced stage compared with the group negative for either mutation. Conclusions: This series provides evidence that FNA is a reliable method for preoperative diagnosis of high-risk thyroid nodules. pTERTC228T/BRAFV600E mutations could be a marker of poor prognosis. Its use as a personalized molecular medicine tool to individualize treatment decisions and follow-up design needs to be further studied.

The BRAFV600E mutation improve diagnostic sensitivity of thyroid nodules with benign or indeterminate cytology results / 中华内分泌外科杂志

Objective To investigate the role of BRAFV600E mutation in diagnosis of thyroid nodules when it is inconsonant with cytological results. Methods This study included 9837 patients who underwent US-FNA. We mainly analyzed 239 cases with benign or indeterminate cytology, but having a detection of BRAFV600E muta-tion. BRAFV600E mutation analysis was performed using a Amplification Refractory Mutation System Polymerase Chain Reaction. Results In 93 nodules with benign cytology results but positive BRAFV600E mutation, 84 nodules were malignant. Based on the results, US-FNA combined with BRAFV600E mutation analysis will improve sensitivity (Se=94.03％)and negative predictive value (NPV=2.69％) of the thyroid nodules diagnosis than using US-FNA alone(Se=71.03％, NPV=20.76％). Conclusion BRAFV600E mutation analysis is an important tool in the diagnosis of PTC with high sensitivity and NPV. When facing patients with benign or indeterminate cytology but positive BRAFV600E mutation, thyroidectomy should be considered.

Correlation of routine ultrasound features and BRAFV600E gene to the cervical lymph node metastasis of thyroid papillary carcinoma / 解放军医学杂志

Objective: To investigate the correlation of routine ultrasound features and BRAFV600E gene to the cervical lymph node metastasis (CLNM) of thyroid papillary carcinoma (PTC). Methods: A hundred and seven patients with thyroid papillary carcinoma who underwent thyroid gland resection and cervical lymph node dissection in the First Medical Center of Chinese PLA General Hospital from January to December 2018 were recruited as study subjects, including 56 patients in the metastatic group and 51 patients in the non-metastatic group. Ultrasound examination and BRFV600E gene detection were performed, and the indicators obtained were compared between the two groups including gender, age, focus position (upper middle or inferior), close to the capsule or to the isthmus, focus located in the lef or right lobes, single or multiple, maximum diameter, aspect ratio, boundary, shape, internal echo, internal components, peripheral halos, calcification, microcalcification, color Doppler features, Hashimoto's disease, and BRFV600E gene mutation. Logistic regression analysis was conducted for gender, age, close to the capsule or to the isthmus, maximum diameter, shape, calcification, microcalcification, color Doppler features, Hashimoto's disease, and BRAFV600E gene mutation to establish a predictive model of cervical lymph node metastasis in patients with thyroid papillary carcinoma, and the predictive efficacy of the model was evaluated. Results: The results compared between the two groups showed that statistically significant differences existed in gender, age, maximum diameter of the lesion, close to capsule, close to isthmus, shape, acoustic halo, calcification and microcalcification (P<0.05). Logistic multivariate regression analysis showed that gender (OR=4.197), age (OR=4.895) and maximum diameter (OR=3.636) were risk factors for cervical lymph node metastasis in patients with thyroid papillary carcinoma (P<0.05). The prediction model established by logistic regression analysis was logistic (P)=-21.957+1.434×gender+1.588×age+1.291×maximum diameter. The efficiency test results of the prediction model showed that the sensitivity and specificity were up to 75.00% and 72.55% respectively. Conclusions: It is preliminarily confrmed that BRFV600E gene mutation is not correlated with cervical lymph node metastasis in patients with thyroid papillary carcinoma. For male patients aged less than 55 years old, the risk of cervical lymph node metastasis should be on guard when the maximum diameter of the carcinoma is large than 10 mm.

Screening of monomers from traditional Chinese medicine for specifically inhibiting brafV600E CT26 cells / 中国药理学通报

Aim: To screen BRAFV600E CT26 cell inhibitors from monomers of traditional Chinese medicine (TCM). Methods CT26 cell line was constructed with lentivirus plasmid to stably express BRAFV6C0E. The proliferation, migration and expression of related proteins in MEK/ERK signaling pathway were detected. The monomers of TCM were detected for biological activities as potential BRAF inhibitors by Discovery Studio 4. 0, and further evaluated by MTT assay. Results: The proliferation and migration of BRAFV6C0E CT26 cells were obviously strengthened compared with wild type control. The expressions of proteins in MEK/ ERK pathway were also activated in BRAFV6C0E CT26 cells. Compared with wild type control, Aloin, Angoroside C and Cyasterone exhibited the potent effect against BRAFV600E in CT26 cells (P <0. 05), and could down-regulate the expression of BRAFV600E. Conclusion: Aloin, Angoroside C, Cyasterone might be the potent inhibitors against BRAF for colon treatment.

The BRAFV600E mutation improve diagnostic sensitivity of thyroid nodules with benign or indeterminate cytology results / 中华内分泌外科杂志

Objective@#To investigate the role of BRAFV600E mutation in diagnosis of thyroid nodules when it is inconsonant with cytological results.@*Methods@#This study included 9837 patients who underwent US-FNA. We mainly analyzed 239 cases with benign or indeterminate cytology, but having a detection of BRAFV600E mutation. BRAFV600E mutation analysis was performed using a Amplification Refractory Mutation System Polymerase Chain Reaction.@*Results@#In 93 nodules with benign cytology results but positive BRAFV600E mutation, 84 nodules were malignant. Based on the results, US-FNA combined with BRAFV600E mutation analysis will improve sensitivity (Se=94.03%) and negative predictive value (NPV=2.69%) of the thyroid nodules diagnosis than using US-FNA alone (Se=71.03%, NPV=20.76%) .@*Conclusion@#BRAFV600E mutation analysis is an important tool in the diagnosis of PTC with high sensitivity and NPV. When facing patients with benign or indeterminate cytology but positive BRAFV600E mutation, thyroidectomy should be considered.

BRAFV600E mutation in hairy cell leukemia: A single-center experience

Background: BRAFV600E mutation has been reported as a unique genetic lesion of hairy cell leukemia (HCL), a subset of which lacks this lesion and shows adverse outcomes. Aims: To determine the prevalence of BRAFV600E in HCL from our center and derive clinicopathological correlation, if any. Materials and Methods: A 9-year retrospective analysis of 46 consecutive cases of HCL diagnosed on morphology and immunophenotyping was done. Stained smears were used as samples for amplification refractory mutation system polymerase-chain reaction using fluorescent primers for mutation detection. Results: BRAFV600E mutation was detected in 41/46 patients (89.1%) while absent in control samples of chronic lymphocytic leukemia. Cases mimicking HCL-variant clinically or immunophenotypically too showed the presence of this mutation. HCL with mutated BRAF presented at a younger age. No statistical difference in blood counts, tumor load, and immunophenotype patterns existed among BRAF mutated and unmutated group. Nine patients (45%) with mutated BRAF had residual disease following treatment with cladribine. Conclusion: BRAFV600E mutation analysis has a definitive role in the diagnosis of HCL.

Consistency and clinicopathologic significance of BRAFV600E protein expression and genic mutation in papillary thyroid carcinoma / 临床与实验病理学杂志

Purpose To investigate the consistency and clinicopathologic correlation of BRAFV600E protein expression and gene mutation in papillary thyroid carcinoma. Methods BRAFV600E protein expression and genn mutation was detected respectively by immunohistochemistry of SP and real time-PCR, then the consistency between the both methods was analyzed by Kappa-test, the correlation between BRAFV600E and clinicopatho-logic parameters was analyzed by Chi-square test in papillary thyroid carcinoma. Results The gene mutation and protein expression rates of BRAFV600E were 89.3％ and 88.3％, respec-tively, the differences were not significant, the concordance rate of the both methods was 97.0％, Kappa value was 0.847, the consistence was higher, meanwhile the mutation rates between age ＜45 and ≥45 were respectively 96.8％ and 85.9％, there were significant differences, the positive rates of the both detec-tion methods were higher in thyroid capsule invaded group than non-invaded group, the differences were significant. Conclusion The both methods have higher consistency, the immunohisto-chemistry can be used as an initial screening tool for detecting gene mutation, the gene mutation of BRAFV600E is significantly associated with age and capsule invasion, the relationship is not found between BRAFV600E mutation and the other clinicopatholog-ic parameters.

Analysing the risky factors of central lymph node metastasis in papillary thyroid microcarcinoma / 国际外科学杂志

Objective To evaluate the relationship between clinicopathological features and central lymph nodes metastasis in patients with papillary thyroid microcarcinoma (PTMC),and aim to provide an appropriate operation in clinic.Methods The clinical data of 536 patients undergoing surgical treatment in Department of Thyroid Surgery,the First Affiliated Hospital of Zhengzhou University from January 2015 to December 2017,who had pathologically confirmed popillary thyroid microcarcinoma (PTMC) were retrospectively analyzed.Thyroidectomy and isthmectomy with central compartment neck dissection were performed in all patients.The patients were divided into two groups according to whether there existed central lymph node metastasis,and compared the differences of gender,age,number of cancer lesions,tumor breakthrough envelope,BRAFV600E gene mutation,and tumor maximum diameter in the central lymph node metastasis,respectively,by t-test x2-test.Univariate logistic regression analysis and multivariate binary logistic regression analysis were conducted to find risky factors.Results There existed difference between two groups by completing the gender,age,the BRAFV600E gene mutation and maximal tumor diameter(P ＜ 0.05).Univariate logistic regression analysis indicated that gender (P =0.046),age (P ＜ 0.01),maximal tumor diameter(P ＜0.01) and the BRAFV600E gene mutation(P =0.016) were significant predictors for central lymph nodes metastasis.And multivariate binary logistic regression analysis revealed that the rate of lymph node metastasis significantly increased in cases of larger tumor diameter(P ＜0.01),BRAFV600E gene mutation(P =0.035) and ageing below 45 years old (P ＜ 0.01).Conclusions The treatment for central lymph node metastasis of PTMC should be different considering elements including BRAF600E gene mutation prophylactic,ageing below 45 years old and larger tumor diameter.Therefore central lymphadenectomy should be performed when the primary lesion was resected.

Evidence-based treatment for a patient with advanced rectal cancer with BRAF V600E mutation / 肿瘤

Objective: Based on evidence-based clinical evidence, an appropriate individualized treatment strategy was established for one patient with V-raf murine sarcomarosoncovirus oncogene homologous B (BRAF) V600E-mutant recurrent metastatic rectal cancer after chemotherapy. Methods: After evaluating the case data completely, the treatmentrelated clinical problems were put forward. Cochrance Library, PubMed, CBM (SinoMed), Embase, Chinese Journal Full-Text Database and Wanfang database were retrieved by computer to find the randomized controlled trial, systematic review and Meta-analysis about metastatic rectal cancer with BRAF V600E mutation. The 2001 Oxford Centre for Evidence-based Medicine Levels of Evidence was used to grade the literatures. Results: A total of 14 relevant studies were retrieved, most of which were grade A evidence, with high evidence intensity and reliable conclusion. Based on evidence-based clinical evidence, a suitable individualized treatment strategy was established for this patient, which was antiepidermal growth factor receptor (EGFR) antibody combined with BRAF inhibitor treatment. The short-term incidence rate of adverse reactions was low, the clinical symptoms were well controlled, and the result of response evaluation was partial response. Conclusion: Based on evidence-based clinical evidence, a reasonable treatment plan for patients with BRAF V600E-mutant metastatic colorectal cancer can effectively improve the therapeutic effect and prognosis.

Relationship between expression of BRAFV600E and Ki67 protein and clinical pathologic characteristics of thyroid carcinoma / 中国基层医药

Objective@#To explore the relationship between BRAFV600E, Ki67 protein and thyroid carcinoma with different pathologic characteristics, thus to provide clinical evidence on early prognosis and personalized treatment in patients with thyroid carcinoma.@*Methods@#From January 2015 to January 2017, 76 patients diagnosed as papillary thyroid carcinoma(PTC), who treated in Yinzhou People's Hospital, were enrolled.Twenty cases with normal tumor-adjacent tissue after operation and 34 patients with non-PTC were also enrolled as control.All the patients enrolled were not treated with endocrine, chemical and radiotherapy.The expression of BRAFV600E and Ki67 protein was detected by immunohistochemical method, and the BRAFV600E and Ki67 expression and their correlation with different pathologic characteristic of thyroid carcinoma was analyzed by SPSS 18.0.@*Results@#The positive rate of BRAFV600E and Ki67 protein in 110 patients with thyroid carcinoma was 56.36%(62 cases) and 39.09%(43 cases), with the highest expression of BRAFV600E and the lowest expression of Ki67 in patients with PTC respectively.The expression of BRAFV600E and Ki67 protein was all negative in normal tumor-adjacent tissue.The positive expression of BRAFV600E and Ki67 in patients with PTC demonstrated significant difference in TNM stage, tumor side and lymphoid node metastasis(BRAFV600E: χ2=5.281, 9.771, 9.771, all P<0.05; Ki67: χ2 =7.098, 4.070, 5.067, all P<0.05), while with no obvious difference in sex, age and numbers of tumor(BRAFV600E: χ2=0.078, 0.093, 0.061, all P>0.05; Ki67: χ2=0.224, 0.518, 1.281, all P>0.05).@*Conclusion@#The detection of BRAFV600E and Ki67 is benefit on judging of innocent and malignant, the malignant degree of thyroid tissue.The high expression of BRAFV600E and Ki67 is benefit on differentiating PTC and early evaluation of prognosis.

Comparison of orthotropic models of RET/PTC1 rearrangement and BRAFV600E mutation papillary thyroid cancer cell lines in nude mice / 中华内分泌代谢杂志

Objective To observe and compare the different orthotopic models of papillary thyroid cancer ( PTC) cell lines of RET/PTC1 rearrangement and BRAFV600E mutation in nude mice. Methods Human PTC cell lines TPC-1, BHP5-16 and BHP2-7 were used. The genotypes of RET/PTC1 rearrangement and BRAFV600E mutation were determined by realtime-PCR and DNA sequencing analysis. The cells(2×105) were injected into the thyroid gland of nude mice. The nude mice were executed at 4th, 12th week, and then their thyroid tumors were removed and weighed. The levels of thyroid hormone were detected using chemiluminescent immunoassay. Results Both TPC-1 and BHP2-7 cells were identified as RET/PTC1 rearrangement by real time-PCR, and the expression of RET/PTC1 rearrangement in BHP2-7 cell was higher than that of TPC-1 cell. BRAFV600E mutation was found in BHP5-16 cell by DNA sequencing analysis, but was not found in TPC-1 and BHP2-7 cells. There were different characteristics in three orthotopic nude model groups. Tumorigenic rates of TPC-1 and BHP5-16 groups were 100%, but the growth of tumor was more rapid in BHP5-16 group than that in TPC-1 group, with more weight tumor. The changes of thyroid hormone levels in BHP5-16 group and TPC-1 group were the same, which were normal at 4th week and sharply decreased at 12 th week(P0. 05). Conclusions It showed difference in the orthotopic models of PTC cell lines of RET/PTC1 rearrangement and BRAFV600E mutation in nude mice. BRAFV600E mutation has obvious impacts on increasing tumorigenic rate and promotion of tumor growth in the orthotopic model. It should not be ignored that advanced thyroid tumor will lead to the destruction of thyroid function.

Comparison of diagnostic value of TIRADS, BSRTC, BRAFV600E mutation detection and their combined use in differentiating thyroid nodules / 中华内分泌代谢杂志

Objective To compare the diagnostic efficiency of the thyroid imaging reporting and data system (TIRADS), the Bethesda system for reporting thyroid cytopathology(BSRTC) and BRAFV600E detection, and their combined use in the differentiation between benign and malignant thyroid nodules. Methods One hundred and twenty eight patients with 128 thyroid nodules who were scheduled for ultrasound-guided fine-needle aspiration biopsy (FNAB) were recruited for the study. All of them underwent ultrasound, fine-needle aspiration cytology(FNAC) examination, and BRAFV600E detection. TIRADS and BSRTC systems were adopted to judge the ultrasound and FNAC results. The receiver operating characteristic curve was established to assess the diagnostic value of each method. Results The sensitivity, specificity, and AUC of TIRADS were 74. 3%, 84. 5%, and 0. 794, respectively. BSRTC had higher specificity(98. 3%) and equal sensitivity compared to TIRADS. The sensitivity, specificity, and AUC of BRAFV600E detection were the highest ones among the three methods. Combinations of different methods could increase the diagnostic sensitivity and accuracy. The combination of FNAC and BRAFV600E detection significantly increased the diagnostic efficiency(AUC=0. 984), with sensitivity 98. 6% and specificity 98. 3%. Conclusions The diagnostic value of BRAFV600E detection in the differentiation of benign and malignant thyroid nodules is better than both TIRADS and BSRTC, and the combination of FNAC and BRAFV600E detection reaches the best diagnostic efficiency.

Effects of BRAF(V600E) mutation on Na(+)/I(-) symporter expression in papillary thyroid carcinoma / 华中科技大学学报（医学）（英德文版）

Radioiodine ablation (RIA) therapy is one of the most important treatments for papillary thyroid carcinoma (PTC), but some patients who received (131)I have radioiodine-refractory disease caused by the decreased expression of the Na(+)/I(-) symporter (NIS). BRAF(V600E) mutation is one possible risk factor that can disturb the NIS expression, but the roles are unclear in clinical practice. This research discussed the association of BRAF(V600E) mutation and NIS expression in PTC tissue and the clinical implications in RIA therapy. 134 PTC samples were collected between June 2013 and June 2014 from Tongji Hospital affiliated to Tongji Medical College, and their clinical characteristics were analyzed. RT-PCR was used to detect the BRAF(V600E) mutation from formalin-fixed paraffin-embedded samples, and immunohistochemistry was applied to detect the NIS expression. IPP software was used to calculate the relative expression quantity of NIS. We found that there was no significant correlation between the absorbance (A) values of NIS and clinicopathologic features in these cases, even thyroid stimulating hormone. BRAF(V600E) mutation showed inhibitory effect on the NIS expression without statistically significant difference in all PTC cases (β=-0.0195, P=0.085), but in the subgroup without hashimoto's thyroiditis (HT), BRAF(V600E) mutation could significantly inhibit the NIS expression (β=-0.0257, P=0.046). The results indicate that BRAF(V600E) mutation is correlated with a lower expression of NIS in PTCs without HT, suggesting the radioiodine-refractory effects during RIA therapy in these patients.

Expression of BIM gene in papillary thyroid carcinoma and its association with the BRAFV600E mutation / 中华内分泌代谢杂志

To study the link between BRAFV600E status and the expression of BIM gene in papillary thyroid carcinoma( PTC) tissues and to analyze the association of these factors with clinicopathological characteristics. BRAFV600E status was determined by MASA-PCR, and qPCR was applied to detect the expression of BIM gene. Finally, the associations of these factors with clinicopathological characteristics were analysed. The rate of mutant BRAFV600E in PTC was 54. 1% , and the expression of BIM gene was lowered in BRAFV600E positive PTC tissues. Additionally, there was significant association( P < 0. 05) between BRAFV600E positiveness and raised TNM Staging (Ⅲ/ Ⅳ), and lowered BIM expression was significantly associated (P<0. 05) with the tumor size and raised TNM Staging(Ⅲ/ Ⅳ). These findings may help us to know more about the mechanism of PTC and to develop new diagnostic biomarkers or prognostic indicators of PTC.

Analysis of the factors associated with clinicopathological characteristics of papillary thyroid carcinoma / 中华内分泌代谢杂志

Objective BRAFV600E mutation, RET/PTC rearrangement, and the concomitant of Hashimoto's thyroiditis(HT) could influence clinicopathological features of papillary thyroid carcinoma (PTC).This study is to investigate the distribution of three factors in PTC and to analyze their associations with clinicopathological characteristics.Methods Fine-needle aspiration samples were collected in a total of 122 conventional PTC patients, who were confirmed by surgery.The clinicopathological features were collected to analyze its association with different factors.BRAFV600E mutation and RET/PTC rearrangement were detected by pyrosequencing and Taqman-qPCR, respectively.Results BRAFV600E mutation was significantly correlated with an older age and a less coexistence with HT(P＜0.05).In contrast, RET/PTC rearrangement was more prevalent in young patients and was associated with the concomitant of HT(P＜0.05).In the age of ≥20 year and＜45 year groups, BRAFV600E mutation was significantly associated with extrathyroidal invasiveness.RET/PTC rearrangement was significantly associated with bilateral lymph node metastasis and the number of metastatic lymph node.Conclusions The distribution of three factors were different in PTC patients.In addition to the age at diagnosis, all of three factors should also be considered together to analyze the association of clinicopathological features of PTC.

Detection of BRAFV600E mutation with central compatment lymphy node metastasis in cN0 papillary thyroid carcinoma / 实用医学杂志

reference index of lymph node in central region.

Recent advances in molecular targeted therapy and expression of iodine uptake relative protein in papillary thyroid carcinoma with BRAFV600E mutation / 中华内分泌代谢杂志

Type-B RAF (BRAF) gene is one of the most popular genes of thyroid carcinoma in recent studies,and its mutation has significant relationships with the occurrence,development,treatment,and prognosis of papillary thyroid carcinoma(PTC).The influence of BRAFV600E mutation on the expression of iodine uptake relative proteins in PTC cells and the value of molecular targeted therapy with BRAFV600Einhibitors in the clinical treatment of PTC in the future were summarized in this review.