Association of BRAF V600E mutation with tumor recurrence in a small sample of Filipino patients with papillary thyroid cancer in a single center

Background and Objective@#Epidemiological studies have shown that Filipinos have a higher prevalence of welldifferentiated thyroid cancer and a higher rate of recurrence. The BRAF V600E mutation has been proposed as a potential prognostic marker in aggressive papillary thyroid cancers. In this study, we determined whether this mutation is a risk factor for tumor recurrence in papillary thyroid cancer among Filipinos.@*Methods@#We conducted an age and sex-matched case-control study of patients with papillary thyroid cancer; we had two groups – with and without tumor recurrence – of 14 patients each, with at least a 5-year follow-up. We extracted the DNA samples from the patients’ (paraffin-embedded) tumor biopsy tissue blocks from thyroidectomy specimens, then detected the BRAF V600E mutation using polymerase chain reaction. The McNemar’s test for difference of proportions in paired data was used to determine the association of BRAF V600E mutation with recurrence. @*Results@#The BRAF V600E mutation was found in 57.14% of all cases. We found a prevalence of 64.29% among those with recurrence and 50.00% among those without recurrence, with no significant difference between the two groups (p = 0.688).@*Conclusion@#Our study showed the BRAF V600E mutation was not associated with recurrence. We encountered several limitations: we had limited data regarding molecular methodologies in the Philippine setting, we had a small sample size, and therefore we could not study other parameters (e.g., tumor characteristics, lymph node metastasis, stage of disease). We hope that this paves the way for future studies and collaborations to establish the role of BRAF V600E in Filipinos with papillary thyroid tumor recurrence.

Characterization of BRAF mutation in patients older than 45 years with well-differentiated thyroid carcinoma / Caracterização da mutação em BRAF em pacientes acima de 45 anos com carcinoma bem diferenciado de tireoide

Abstract Introduction: Papillary thyroid carcinoma is the most frequent endocrine neoplasia and its incidence has tripled over the past 35years. Although papillary thyroid carcinoma carries a good prognosis, 10%-30% of patients still develop recurrence and metastasis. Some clinical and genetic features are associated with worse prognosis. The most frequent mutation is the BRAF p.V600E, which has been associated with many clinical features of poor prognosis. However, many studies have produced controversial results without any association between BRAF mutation and clinicopathological features of poor prognosis. Objective: Since the prognostic value of BRAF mutations remains controversial, this study aims to investigate the importance of this mutation in therapeutic decisions for papillary thyroid carcinoma. Methods: Therefore, we evaluated whether the presence of BRAF mutation is associated with features of poor prognosis in 85 patients with papillary thyroid carcinoma older than 45years treated at A.C. Camargo Cancer Center, from 1980 to 2007. BRAF mutation was evaluated by pyrosequencing. Statistical analysis was performed using SPSS. Results: The mean age of patients was 54 years (range: 45 - 77 years), 73 were women (85.8%) and 12 were men (14.2%). Among them, 39 cases (45.9%) presented extrathyroidal extension and 11 cases had recurrent disease. BRAF mutation was detected in 57 (67%) patients. No significant association was observed between BRAF mutation and gender (p =0.743), age (p = 0.236), N-stage (p =0.423), vascular and perineural infiltration (p =0.085 or multifocality (p = 1.0). Although not statistically significant, the majority of patients with recurrent disease were BRAF positive (9 out of 11) (p =0.325). Patients affected by BRAF mutation are associated with tumors larger than 1 cm (p =0.034) and with extrathyroidal extension (p =0.033). Conclusion: Although BRAF testing is widely available, there are no consistent data to support improvement in outcomes from incorporating it into therapeutic decision for thyroid cancer.

Resumo Introdução: O carcinoma papilífero de tireoide é a neoplasia endócrina mais frequente e sua incidência triplicou nos últimos 35 anos. Embora o carcinoma papilífero de tireoide tenha um bom prognóstico, 1% a 30% dos pacientes desenvolvem recorrência e metástase. Algumas características clínicas e genéticas estão associadas a um pior prognóstico. A mutação mais frequente é a BRAF p.V600E, a qual tem sido associada a muitas características clínicas de pior prognóstico. No entanto, muitos estudos apresentam resultados controversos, sem qualquer associação entre a mutação em BRAF e características clinicopatológicas de pior prognóstico. Objetivo: Uma vez que o valor prognóstico das mutações em BRAF permanece controverso, investigar a importância dessa mutação em decisões terapêuticas para o carcinoma papilífero de tireoide. Método: Foi avaliada a associação da mutação em BRAF com características de pior prognóstico em 85 pacientes com carcinoma papilífero de tireoide acima de 45 anos tratados no A.C. Camargo Cancer Center, de 1980 a 2007. A mutação em BRAF foi avaliada por pirossequenciamento. A análise estatística foi feita com o software SPSS. Resultados: A média de idade dos pacientes foi de 54 anos (variação de 45 - 77), 73 eram mulheres (85,8%) e 12 eram homens (14,2%). Entre eles, 39 casos (45,9%) apresentaram extensão extratireoidiana e 11, doença recorrente. A mutação em BRAF foi detectada em 57 (67%) pacientes. Não foi observada associação significante entre mutação em BRAF e sexo (p = 0,743), idade (p = 0,236), estágio N (p = 0,423), infiltração vascular e perineural (p = 0,085) ou multi-focalidade (p = 1,0). Apesar de não apresentar associação estatística, a maioria dos pacientes com doença recorrente foi positiva para BRAF (9 em 11) (p = 0,325). Os pacientes afetados pela mutação em BRAF estão associados a tumores maiores do que 1 cm (p = 0,034) e com extensão extratireoidiana (p = 0,033). Conclusão: Embora a mutação em BRAF seja amplamente avaliada, não há dados consistentes que demonstrem uma melhor sobrevida ou benefício clínico ao incorporá-la à decisão terapêutica para o câncer de tireoide.

The BRAFV600E mutation analysis and risk stratification in papillary thyroid carcinoma

ABSTRACT Objective: Although the prognostic role of BRAFV600E mutation in papillary thyroid carcinoma (PTC) is controversial, the American Thyroid Association (ATA) includes the mutational status in their risk stratification system. To evaluate the impact of the BRAFV600E mutation status on PTC risk stratification. Subjects and methods: PTC patients attending a university-based hospital who had the analysis of the BRAFV600E mutation were included. Persistent disease was defined as the presence of biochemical or structural disease. The performance of the ATA risk stratification system on predicting persistent disease with or without the BRAFV600E mutation status information was evaluated. Results: Of the 134 patients evaluated, 44 (32.8%) carried BRAFV600E mutation. The median tumor size was 1.7 cm (P25-75 1.0-3.0), 64 (47.8%) patients had lymph node, and 11 (8.2%) distant metastases. According to the ATA risk stratification system, patients were classified as low, intermediate, and high risk in 55 (41%), 59 (44%), and 20 (14%) patients, respectively. The data on BRAFV600E mutation reclassified 12 (8.9%) patients from low to intermediate risk. After a median follow-up of 8.5 years, the prevalence of persistent disease was similar in patients with and without BRAFV600E mutation (P = 0.42). Multivariate analysis failed to demonstrate an association between the BRAFV600E mutation and persistent disease status (RR 0.96; 95%CI 0.47-1.94). Notably, none of the patients reclassified from low to intermediate risk showed persistent disease on follow-up. Conclusion: Inclusion of BRAFV600E mutational status has a limited impact on risk stratification and does not add to the prediction of outcomes in PTC patients.

Clinicopathological spectrum of hairy-cell leukemia: A single-center study with brief review of Indian literature

Objective: The presence of specific chemotherapeutic protocols for hairy cell leukemia (HCL) makes it essential to discriminate this entity from other lymphoproliferative disorders. Hence, awareness of the variations in clinical presentations and immunophenotypic aberrancies is requisite to ensure diagnostic accuracy. Materials and Methods: A retrospective study was carried out to analyze the clinical-pathological profile of patients with HCL diagnosed over a period of 81 months (2010–September 2017) in our institute. Flow cytometry was performed in all the patients, and further, BRAFV600E mutation analysis was performed by real-time polymerase chain reaction in a limited number of samples. Result: A total of 353 lymphoproliferative disorders were assessed during the period, of which 16 (4.5%) were diagnosed as HCL, which included 15 cases of classical HCL and single case of HCL-v. Striking male predominance was noted with a median age of 52 (range 22–90 years). 47% patients presented with pancytopenia, while 20% cases had leukocytosis. Three patients presented with bleeding diathesis in the form of melena and purpuric spots. The absence of splenomegaly was observed in 20% patients (4/15) while 2 (13.3%) cases had lymphadenopathy. Hypocellular marrow was observed in 13% cases. Bright expression of CD20/CD22 along with CD25/CD103/CD123/CD11c was noted in all the patients of classical HCL. Aberrant expression of CD23 and CD5 was seen in 33% ( n =5) and 6.7% ( n =1) cases respectively. CD200 was positive in all the 5/15 cases tested. The case of HCL–v presented with very high leukocyte count and exhibited a CD103/CD11c+ and CD123/CD25- profile. BRAFV600E, mutation was present in all the four patients tested who included patients with a hypocellular marrow and absent splenomegaly. Conclusion: HCL has characteristic profiles, yet it may exhibit unusual clinical and immunophenotypic presentations. Perspicacious use of flow cytometry and BRAFV600E mutation analysis will aid in the diagnosis in unprecedented cases

Contrast-enhanced ultrasound and BRAF mutation in diagnosis of extracapsular extension of papillary thyroid carcinoma / 中国医学影像技术

Objective: To investigate the value of contrast-enhanced ultrasound (CEUS) and BRAF mutation in pre-operative diagnosis of extracapsular extension in papillary thyroid carcinoma (PTC). Methods: A total of 129 PTC in 119 patients were enrolled, including 25 PTC with extracapsular extension (A group) and 104 PTC without extracapsular extension (B group). Both the range of contact between PTC and thyroid capsule and the presence of discontinued capsule in ultrasound (US) and CEUS, as well as BRAF mutation examination result were recorded. The diagnostic performance of the above features in diagnosis of extracapsular extension in PTC, and the overall diagnostic performance of their combination were calculated. Results: Significant differences were found in BRAF mutation, range of capsule contact and the presence of discontinued capsule between US and CEUS (all P<0.05). In different ranges of contact, the threshold of ≥25% demonstrated the highest diagnostic accuracy in both US and CEUS. CEUS had higher accuracy than US in diagnosis of extracapsular extension in PTC based on single ultrasonic feature as well as the combination of ultrasonic features and BRAF mutation. The accuracy of CEUS combined with BRAF was 88.37% (114/129). Conclusion: The characteristics of extracapsular extension in PTC include BRAF mutation, ≥25% range of contact between PTC and thyroid capsule and the presence of discontinued capsule in US and CEUS. CEUS has higher accuracy than US. Combination of BRAF mutation and CEUS has important clinical significance in preoperative diagnosis of PTC extracapsular extension.

Association between BRAF (V600E) mutation and clinicopathological features of papillary thyroid carcinoma: a Brazilian single-centre case series

ABSTRACT Objectives: We aimed to investigate the prevalence of the BRAF (V600E) mutation in consecutive cases of papillary thyroid carcinoma (PTC) in patients diagnosed and treated at the Hospital Sao Rafael (Salvador, BA, Brazil) and evaluate its association with clinical and pathological characteristics of PTC. Subjects and methods: We retrospectively enrolled in the study a total of 43 consecutive PTC patients who underwent total thyroidectomy. We performed DNA extraction from formalin-fixed paraffin-embedded (FFPE) tumour tissue samples. Polymerase chain reaction (PCR) and direct sequencing were used to determine BRAF (V600E) mutation status. Univariate and multivariate logistic regression analyses were employed to identify independent associations. Results: The prevalence of BRAF (V600E) mutation was 65.1% (28/43). A high frequency of older patients (p value: 0.004) was observed among the BRAF-mutated PTC group and, in contrast, a low frequency of concurrent Hashimoto's thyroiditis (HT) (p value: 0.011) was noted. Multivariate analysis confirmed that older age (OR: 1.15; 95% CI: 1.00 - 1.33; p value: 0.047) and HT (OR: 0.05; 95% CI: 0.006-0.40; p value: 0.005) were independent factors associated with BRAF (V600E) mutation. Conclusion: We found a high prevalence of BRAF (V600E) mutation in PTC cases. Older age and no concurrent HT were independently associated with BRAF (V600E) mutation.

Expression of NF2 Modulates the Progression of BRAFV600E Mutated Thyroid Cancer Cells

BACKGROUND: We previously reported the frequent neurofibromatosis 2 (NF2) gene mutations in anaplastic thyroid cancers in association with the BRAF V600E mutation. We aimed to investigate the role of NF2 in thyroid cancer with BRAF mutation. METHODS: To identify the function of NF2 in thyroid cancers, we investigated the changes in cell proliferation, colon formation, migration and invasion of thyroid cancer cells (8505C, BHT101, and KTC-1) with BRAF V600E mutation after overexpression and knock-down of NF2. We also examined how cell proliferation changed when NF2 was mutagenized. Human NF2 expression in papillary thyroid carcinoma (PTC) was analyzed using the The Cancer Genome Atlas (TCGA) data. RESULTS: First, NF2 was overexpressed in 8505C and KTC-1 cells. Compared to control, NF2 overexpressed group of both thyroid cancer cells showed significant inhibition in cell proliferation and colony formation. These results were also confirmed by cell migration and invasion assay. After knock-down of NF2 in 8505C cells, there were no significant changes in cell proliferation and colony formation, compared with the control group. However, after mutagenized S288* and Q470* sites of NF2 gene, the cell proliferation increased compared to NF2 overexpression group. In the analysis of TCGA data, the mRNA expression of NF2 was significantly decreased in PTCs with lateral cervical lymph node (LN) metastasis compared with PTCs without LN metastasis. CONCLUSION: Our study suggests that NF2 might play a role as a tumor suppressor in thyroid cancer with BRAF mutation. More studies are needed to elucidate the mechanism how NF2 acts in thyroid cancer with BRAF mutation.

Active and Passive Smoking, BRAF(V600E) Mutation Status, and the Risk of Papillary Thyroid Cancer: A Large-Scale Case-Control and Case-Only Study / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: The association between tobacco smoking and thyroid cancer remains uncertain. We evaluated the associations of active and passive smokingwith the risk of papillary thyroid cancer (PTC), the most common type of thyroid cancer, and with the BRAF(V600E) mutation, the most common oncogenic mutation in PTC related to poor prognosis. MATERIALS AND METHODS: We conducted this study with newly diagnosed PTC patients (n=2,142) and community controls (n=21,420) individually matched to cases for age and sex. Information on active and passive smoking and potential confounders were obtained from structured questionnaires, anthropometric measurements, and medical records. BRAF(V600E) mutation status was assessed in PTC patients. We evaluated the associations of active and passive smoking with PTC and BRAF(V600E) mutation risk using conditional and unconditional logistic regression models, respectively. RESULTS: We did not find associations between exposure indices of active and passive smoking and PTC risk in both men and women, except for the association between current smoking and lower PTC risk. Cumulative smoking ≥ 20 pack-years was associated with lower BRAF(V600E) mutation risk in male PTC patients (odds ratio [OR], 0.55; 95% confidence interval [CI], 0.30 to 1.00). The CI for the association was wider in female PTC patients (OR, 0.23; 95% CI, 0.02 to 2.62), possibly owing to a smaller sample size in this stratum. CONCLUSION: We did not find consistent associations between active and passive smoking and PTC risk. Cumulative smoking ≥ 20 pack-years was associated with lower BRAF(V600E) mutation risk in male PTC patients.

Utility of BRAF VE1 Immunohistochemistry as a Screening Tool for Colorectal Cancer Harboring BRAF V600E Mutation

BACKGROUND: BRAF mutation has been recognized as an important biomarker of colorectal cancer (CRC) for targeted therapy and prognosis prediction. However, sequencing for every CRC case is not cost-effective. An antibody specific for BRAF V600E mutant protein has been introduced, and we thus examined the utility of BRAF VE1 immunohistochemistry for evaluating BRAF mutations in CRC. METHODS: Fifty-one BRAF-mutated CRCs and 100 age and sexmatched BRAF wild-type CRCs between 2005 and 2015 were selected from the archives of Asan Medical Center. Tissue microarrays were constructed and stained with BRAF VE1 antibody. RESULTS: Forty-nine of the 51 BRAF-mutant CRCs (96.1%) showed more than moderate cytoplasmic staining, except for two weakly stained cases. Six of 100 BRAF wild-type cases also stained positive with BRAF VE1 antibody; four stained weakly and two stained moderately. Normal colonic crypts showed nonspecific weak staining, and a few CRC cases exhibited moderate nuclear reactivity (3 BRAF-mutant and 10 BRAF wild-type cases). BRAF-mutated CRC patients had higher pathologic stages and worse survival than BRAF wild-type patients. CONCLUSIONS: BRAF VE1 immunohistochemistry showed high sensitivity and specificity, but occasional nonspecific staining in tumor cell nuclei and normal colonic crypts may limit their routine clinical use. Thus, BRAF VE1 immunohistochemistry may be a useful screening tool for BRAF V600E mutation in CRCs, provided that additional sequencing studies can be done to confirm the mutation in BRAF VE1 antibody-positive cases.

The Significance of TROP2 Expression in Predicting BRAF Mutations in Papillary Thyroid Carcinoma

BACKGROUND: Trophoblast antigen 2 (TROP2) is a human trophoblast cell-surface glycoprotein that is overexpressed in several types of epithelial cancers, and is suggested to be associated with an unfavorable prognosis. BRAF mutations are the most common genetic alteration in papillary thyroid carcinoma (PTC). We evaluated the correlation between TROP2 expression and BRAF mutation in PTC. METHODS: First, we carried out pyrosequencing for BRAF mutations and immunohistochemistry for TROP2 expression with a tissue microarray consisting of 52 PTC cases. Membranous staining in at least 5% of tumor cells was designated as positive staining and we analyzed the relationship between TROP2 expression and diverse clinicopathological factors, including BRAF mutation. Second, we tested TROP2 mRNA expression in three thyroid cancer cell lines with BRAF mutations (BCPAP, SNU790, and 8505C) and a normal thyroid cell line. Additionally, we checked TROP2 protein levels in a normal thyroid cell line after introduction of the BRAF V600E mutation. RESULTS: In this study, 21 of 26 cases with BRAF mutation showed TROP2 immunoreactivity, whereas all 26 cases without BRAF mutation showed no immunoreactivity for TROP2 with a statistically significant difference (p<.001). Upregulation of TROP2 mRNA was observed in all three thyroid cancer cell lines, but not in the normal thyroid cell line. Interestingly, however, the TROP2 expression was increased in the normal thyroid cell line after introduction of the BRAF V600E mutation. CONCLUSIONS: Based on these results, we concluded that TROP2 expression is significantly associated with BRAF mutation and that TROP2 immunohistochemistry could be used for predicting BRAF mutations or diagnosing papillary thyroid carcinoma.

Extent of field change in colorectal cancers with BRAF mutation

<p><b>INTRODUCTION</b>Sporadic colorectal cancers with BRAF mutations constitute two distinct subgroups of colorectal cancers. Recent studies have linked the presence of the BRAF mutation to a familial inheritance pattern. This was a proof-of-concept study that aimed to examine: (a) the extent of field change in sporadic colorectal cancers with BRAF mutation; and (b) the extent of resection margins required and the pattern of DNA mismatch repair protein loss in these tumours.</p><p><b>METHODS</b>Eight microsatellite instability-high tumours with positive BRAF mutation from an existing histopathological database were selected for BRAF mutation and mismatch repair protein analysis.</p><p><b>RESULTS</b>All the resection margins were negative for BRAF mutation. Three tumours had loss of MLH1 and PMS2 expressions, and five tumours had no protein loss. Six peritumoral tissues were negative and one was positive for BRAF mutation.</p><p><b>CONCLUSION</b>The results suggest that any early field change effect is restricted to the immediate vicinity of the tumour and is not a pan-colonic phenomenon. Current guidelines on resection margins are adequate for BRAF mutation-positive colorectal cancers. Any suggestion of a hereditary link to these tumours is likely not related to germline BRAF gene mutations. The pattern of protein loss reinforces previous findings for the two subgroups of BRAF mutation-positive colorectal cancers.</p>

The correlation between glucose metabolism and BRAF mutation in 18 F-FDG PET/CT imaging of papil-lary thyroid carcinoma / 实用肿瘤学杂志

Objective The objective of this study was to evaluate the correlation between maximum standard uptake values( SUVmax ) of 18 F-FDG PET/CT and BRAF mutation in patients with papillary thyroid car-cinoma(PTC).Methods Atotalof51patients(meanageof49.3±12.9years)whounderwent18F-FDGPET/CT imaging and biopsy before thyroidectomy were retrospectively analyzed. Based on the pathological results, there were 48 patients with PTC and 3 patients with follicular thyroid carcinoma(FTC). The SUVmax of thyroid nodule was measured by semi-quantitative analysis. The correlation between clinical data( gender,age,tumor size and thyroglobulin concentration) and SUVmax was also analyzed. The difference of SUVmax between the two groups was analyzed by comparing the BRAF V600E mutation group and the non-mutation group. Results In patients with PTC,27 patients had BRAF V600E mutations and 11 patients had no tumor mutation. The SUVmax of BRAF V600E mutation group was significantly higher than that in the non-mutated group(5. 5 ± 3. 9 vs. 2. 2 ± 1. 2,P=0. 002). The SUVmax of patients with tumor diameter≥1 cm was significantly higher than that in patients with tumor <1 cm(P<0. 05). The SUVmax of patients with elevated thyroglobulin concentration was higher than that in normal patients(P<0. 05). No BRAF V600E mutation was observed in the FTC group. Conclusion The BRAF V600E mutant gene has a high SUVmax value in patients with PTC. There were significant difference in SUVmax among different tumor size and serum thyroglobulin concentration.

Predicting Extrathyroidal Extension in Patients With Papillary Thyroid Microcarcinoma According to a BRAF Mutation

OBJECTIVES: The aim of this study was to evaluate the association between preoperative parameters and extrathyroidal extension (ETE) of papillary thyroid microcarcinoma (PTMC) according to the BRAF mutation and to evaluate the preoperative predictability of ETE. METHODS: We analyzed the medical records of 332 patients with PTMC (140 in the BRAF– group and 192 in the BRAF+ group). The presence of ETE was subjected to a correlation analysis with age, sex, tumor size, clinical nodal status, and ultrasonography (US) findings. Among the US findings, the correlation between tumors and the thyroid capsule was categorized into four groups; US group A, intraparechymal; US group B, tumor abutting the capsule 50% of diameter; and US group D, tumor destroyed the capsule. The predictive value of ETE, including sensitivity, specificity, and positive and negative predictive values were evaluated. RESULTS: Tumor size and US group were significantly correlated with gross ETE in the BRAF– and BRAF+ groups. Tumor size of 0.5 cm and US groups B and C in the BRAF– group were cutoff values for gross ETE, with a negative predictive value of 100%, whereas tumor size of 0.7 cm and US groups A and B in the BRAF+ group had negative predictive values of 92.4% and 100%, respectively. CONCLUSION: Excluding of ETE by US was categorized according to tumor size and US findings. A different categorization to exclude ETE is needed according to the BRAF mutation.

Evaluation of the VE1 Antibody in Thyroid Cytology Using Ex Vivo Papillary Thyroid Carcinoma Specimens

BACKGROUND: Recently, VE1, a monoclonal antibody against the BRAFV600E mutant protein, has been investigated in terms of its detection of the BRAFV600E mutation. Although VE1 immunostaining and molecular methods used to assess papillary thyroid carcinoma in surgical specimens are in good agreement, evaluation of VE1 in thyroid cytology samples is rarely performed, and its diagnostic value in cytology has not been well established. In present study, we explored VE1 immunoexpression in cytology samples from ex vivo papillary thyroid carcinoma specimens in order to minimize limitations of low cellularity and sampling/targeting errors originated from thyroid fineneedle aspiration and compared our results with those obtained using the corresponding papillary thyroid carcinoma tissues. METHODS: The VE1 antibody was evaluated in 21 cases of thyroid cytology obtained directly from ex vivo thyroid specimens. VE1 immunostaining was performed using liquid-based cytology, and the results were compared with those obtained using the corresponding tissues. RESULTS: Of 21 cases, 19 classic papillary thyroid carcinomas had BRAFV600E mutations, whereas two follicular variants expressed wild-type BRAF. VE1 immunoexpression varied according to specimen type. In detection of the BRAFV600E mutation, VE1 immunostaining of the surgical specimen exhibited 100% sensitivity and 100% specificity, whereas VE1 immunostaining of the cytology specimen exhibited only 94.7% sensitivity and 0% specificity. CONCLUSIONS: Our data suggest that VE1 immunostaining of a cytology specimen is less specific than that of a surgical specimen for detection of the BRAFV600E mutation, and that VE1 immunostaining of a cytology specimen should be further evaluated and optimized for clinical use.

Use of the BRAF V600E mutation in fine-needle aspirations of thyroid nodules / 中华超声影像学杂志

Objective To evaluate the diagnostic and prognostic value of BRAF V600E mutation screening of ultrasound-guided fine-needle aspiration (FNA)specimens in patients with thyroid nodule. Methods The BRAF V600E mutation status were assessed in FNA specimens of 104 patients with thyroid nodules before operations.The BRAF mutation status,clinical,and pathology records of the patients were reviewed and the associations between these characteristics and papillary thyroid cancer (PTC ) were analyzed.Results Seventy-one PTC and 14 benign thyroid nodules were included in this study.BRAF V600E mutations were found in 57/71 (80%)PTC.All benign thyroid nodules had no BRAF V600E mutation.The sensitivity,specificity,positive predictive value and negative predictive value of BRAF V600E mutations in differentiation between PTC and benign thyroid nodules were 80%,100%,100% and 50%(P < 0.001 ).In 44 patients with PTC who underwent surgery,the central compartment lymph node metastases and extrathyroidal invasion were not significantly different between BRAF-positive and BRAF-negative PTC (P = 0.283 and 0.307 ).Conclusions BRAF V600E mutation may be a potential tool to facilitate ultrasound in diagnosis of PTC.In patients with PTC,the presence of the BRAFV600E mutation was not significantly associated with prognostic factors.

Study on relationship between expression of survivin, galectin-3 and TPO and BRAFV600E mutation in patients with papillary thyroid microcarcinoma and its clinical significance / 中国免疫学杂志

Objective:To detect the expressions of survivin, galectin-3 and thyroid peroxidase in papillary thyroid microcarcinoma ( PTMC) and cancer adjacent normal tissue to explore the clinical significant.The correlations between the expressions of survivin,Gal-3 and TPO with BRAFV600E gene mutation in PTMC were analyzed.Methods: The expressions of survivin,Gal-3 and TPO were measured by immunohistochemical ( IHC ) method in 56 cases of PTMC tissue and adjacent normal tissue; BRAFV600E mutation was detected by PCR amplification and subsequently sequencing.Chi square test was used to analyse the relation in the expression rates of survivin,Gal-3 and TPO and BRAFV600E gene mutation.Results:The survivin and Gal-3 were strongly expressed but TPO was negatively expressed.The survivin and Gal-3 were negative in adjacent normal tissues but TPO was expressed.There were sig-nificant differences in the expression rates of survivin, Gal-3 and TPO between PTMC tissue and adjacent normal tissue (all P0.05).There were no correlation was found between the expressions of survivin,Gal-3,TPO and the BRAFV600E gene mutation in PTMC(all P>0.05).Conclusion: The strong expressions of survivin and Gal-3,the mild expression of TPO and BRAF mutation may be important in the development of PTMC,and the detection of BRAFV600E gene mutation and the expressions of survivin, Gal-3 and TPO could be used to the judgment of pathogenetic condition and prognostic outcomes.

Diagnostic Effectiveness of PCR-based Tests Detecting BRAF Mutation for Treating Malignant Melanoma: A Systematic Review

BACKGROUND: We aimed to conduct a systematic review of previously published material to evaluate the diagnostic effectiveness of PCR-based tests in detecting BRAF mutation. METHODS: Eight Korean databases, including KoreaMed, Ovid-MEDLINE, and Ovid-EMBASE were used to identify relevant published studies. Nine studies describing usage of real-time PCR, dual-priming oligonucleotide (DPO)-multiplex real-time PCR and allele-specific PCR were included in the final assessment. Two reviewers screened all references independently for assessing the quality of the included articles and extracted data. RESULTS: The rate of detection of the BRAF mutations was lower in the Korean population (11.1-17.2%) than that in the Western population (36.7-82.2%). The diagnostic accuracy of the BRAF mutation tests was assessed on the basis of four previous reports, all of which employed real-time PCR on malignant melanoma. In fact, the diagnostic accuracy of real-time PCR was found to be higher than that of sequencing tests (pooled sensitivity, 0.96; pooled specificity, 0.83; and summary receiver operating characteristic area under the curve, 0.99). In addition, we found that there was no publication bias in meta-analysis. The concordance rate of the BRAF mutation tests compared with reference tests was 87.9-98.1%. CONCLUSIONS: Real-time PCR for the detection of the BRAF gene mutation is an effective technology for determining the appropriateness of treatment with BRAF kinase inhibitors in terminal stage cancer as well as metastatic and malignant melanoma.

Can Ultrasound Be as a Surrogate Marker for Diagnosing a Papillary Thyroid Cancer? Comparison with BRAF Mutation Analysis

PURPOSE: We investigated the merit of ultrasound (US) features and BRAF(V600E) mutation as an additional study of cytology and compared the diagnostic performances of cytology alone, cytology with US correlation, cytology with BRAF(V600E) mutation, and a combination of cytology, US, and BRAF(V600E) mutation all together. MATERIALS AND METHODS: This study included 185 patients (mean age, 48.4 years; range 20-77 years) with 191 thyroid nodules who underwent US-guided fine-needle aspiration (FNA) with an additional BRAF(V600E) mutation test. Three radiologists highly experienced in thyroid imaging retrospectively reviewed US images and classified each nodule into two categories (positive for malignancy or negative for malignancy). Interobserver variability (IOV) of US assessment between the three readers was estimated using the generalized kappa statistic of Landis and Koch. We also calculated the diagnostic performances of these studies. RESULTS: There were 131 cases of malignancy (131/191, 68.6%) and 60 cases of benign nodules (60/191, 31.4%). In terms of IOV of US assessment, the generalized kappa value was 0.242, indicating fair agreement was reached. The combination of cytology with BRAF(V600E) showed higher specificity (100%) and positive predictive value (PPV) (100%) compared to the combination of cytology, BRAF(V600E), and US (specificity 28.3%, 66.7%, 68.3%; PPV 74.6%, 86.6%, 86.8%, respectively; p<0.001). However, cytology with BRAF(V600E) showed lower sensitivity (84.7%) than cytology with BRAF(V600E) and US (96.2%, 98.5%, 95.4%, respectively; p<0.001). CONCLUSION: Considering the diagnostic performance and low reproducibility of US, the combination of FNA with BRAF(V600E) is the most reliable and objective method for diagnosing thyroid malignancy.

Can Ultrasound Be as a Surrogate Marker for Diagnosing a Papillary Thyroid Cancer? Comparison with BRAF Mutation Analysis

PURPOSE: We investigated the merit of ultrasound (US) features and BRAF(V600E) mutation as an additional study of cytology and compared the diagnostic performances of cytology alone, cytology with US correlation, cytology with BRAF(V600E) mutation, and a combination of cytology, US, and BRAF(V600E) mutation all together. MATERIALS AND METHODS: This study included 185 patients (mean age, 48.4 years; range 20-77 years) with 191 thyroid nodules who underwent US-guided fine-needle aspiration (FNA) with an additional BRAF(V600E) mutation test. Three radiologists highly experienced in thyroid imaging retrospectively reviewed US images and classified each nodule into two categories (positive for malignancy or negative for malignancy). Interobserver variability (IOV) of US assessment between the three readers was estimated using the generalized kappa statistic of Landis and Koch. We also calculated the diagnostic performances of these studies. RESULTS: There were 131 cases of malignancy (131/191, 68.6%) and 60 cases of benign nodules (60/191, 31.4%). In terms of IOV of US assessment, the generalized kappa value was 0.242, indicating fair agreement was reached. The combination of cytology with BRAF(V600E) showed higher specificity (100%) and positive predictive value (PPV) (100%) compared to the combination of cytology, BRAF(V600E), and US (specificity 28.3%, 66.7%, 68.3%; PPV 74.6%, 86.6%, 86.8%, respectively; p<0.001). However, cytology with BRAF(V600E) showed lower sensitivity (84.7%) than cytology with BRAF(V600E) and US (96.2%, 98.5%, 95.4%, respectively; p<0.001). CONCLUSION: Considering the diagnostic performance and low reproducibility of US, the combination of FNA with BRAF(V600E) is the most reliable and objective method for diagnosing thyroid malignancy.

BRAF mutation may predict higher necessity of postoperative radioactive iodine ablation in papillary thyroid cancer

PURPOSE: The primary aim of the present study was to analyze the association between high-risk clinicopathologic characteristics and the BRAFV600E mutation. METHODS: From March 2010 to September 2012, we performed analysis of the BRAF mutation (assessing V600E point mutation of BRAF gene, exon 15, on chromosome 7q34 by real-time polymerase chain reaction kit) from 499 papillary thyroid carcinoma (PTC) patients who underwent thyroidectomy. We analyzed the relation between the mutation and known clinicopathologic risk factors of PTC. RESULTS: BRAF mutations were found in 353 of 499 patients (70.7%). On univariate analysis, BRAF mutations were more frequently detected in patients with central lymph node metastasis (78.5% vs. 66.7%, P = 0.007) and classic PTC type (71.3% vs. 16.7%, P = 0.011). Patients with one or more aggressive pathologic feature such as lymph node metastasis, multifocality, and extrathyroidal extension showed higher BRAF mutation rate (73.5% vs. 62.3%, P = 0.022). BRAF mutation group showed more aggressive pathologic features, which is considered as higher necessity of radioactive iodine ablation (relative risk, 1.617; P = 0.035). CONCLUSION: This study found that BRAF mutation is associated with classic PTC and central lymph node metastasis and higher necessity of radioactive iodine ablation.