Immunoexpression of Mutated BRAF V600E Protein in Papillary Thyroid Carcinoma
Asian Oncology Research Journal,
Background and Objectives: In papillary thyroid cancer (PTC), BRAF V600E is the most prevalent genetic alteration, and in different populations, its frequency ranges from 29% to 83%. BRAF mutation is mostly detected by DNA-based molecular methods, which are labor-intensive and time-consuming. A method, immunohistochemistry (IHC), was recently introduced to detect BRAF-mutated proteins. This method enables a monoclonal BRAF V600E mutation-specific antibody that can distinguish BRAF V600E from wild-type protein in conventionally processed, formalin-fixed, paraffin-embedded tissue and the mutant protein that was directly visualized in tumour cells in a tissue context. This study aimed to determine the BRAF V600E-mutated protein's immunoexpression in papillary thyroid carcinoma.
Methodology: The study was a laboratory-based cross-sectional descriptive study. A total of 44 histologically proven paraffin-embedded tissue blocks of PTC were collected. Anti-BRAF V600E rabbit monoclonal primary antibody was used for immunohistochemistry on tissue sections, and the staining intensity was scored from 0 to 3 (+): 0, no cytoplasmic staining in tumour cells; 1: faint cytoplasmic staining in over 10% of tumour cells; 2+, moderate cytoplasmic staining in over 10% of tumour cells; 3+, strong cytoplasmic staining in over 10% of tumour cells. Tumour cells with a score of 1+, 2+, or 3+ were considered positive for mutated BRAF V600E immunoexpression, and those with a score of 0 were considered negative.
Results: Among 44 cases, 34 (77.3%) were positive, and 10 (22.7%) were negative for the mutated BRAF V600E protein by IHC staining. In terms of staining intensity, 4 (9.1%), 20 (45.5%), and 10 (22.7%) cases had IHC scores of 1+, 2+, and 3+, respectively. This study reported a high-frequency rate (77.3%) of mutated BRAF protein, similar to the frequency reported in other Asian countries. There was no association between mutated BRAF V600E protein status and either age or gender.
Conclusion: The most effective PTC diagnostic marker is BRAF V600E mutation. The IHC technique using BRAF V600E mutation-specific antibodies is relatively simple and faster and is therefore proposed as the most reliable first-line method for detecting BRAF V600E-mutated proteins.
- immunohistochemistry (IHC)
- BRAF V600E
- papillary thyroid carcinoma
- formalin-fixed paraffin-embedded tissue
- monoclonal antibody
- cytoplasmic staining
How to Cite
Ge J, Wang J, Wang H, Jiang X, Liao Q, Gong Q, Mo Y, Li X, Li G, Xiong W, Zhao J. The BRAF V600E mutation is a predictor of the effect of radioiodine therapy in papillary thyroid cancer. Journal of Cancer. 2020;11(4):932.
Xing M. BRAF mutation in papillary thyroid cancer: pathogenic role, molecular bases, and clinical implications. Endocr Rev. 2007;28(7):742-62.
Liu C, Chen T, Liu Z. Associations between BRAF(V600E) and prognostic factors and poor outcomes in papillary thyroid carcinoma: a meta-analysis. World J Surg Oncol. 2016;14(1):241.
Huang T, Zhuge J, Zhang WW. Sensitive detection of BRAF V600E mutation by Amplification Refractory Mutation System (ARMS)-PCR. Biomark Res. 2013;1(1):3.
Lasota J, Kowalik A, Wasag B, Wang ZF, Felisiak-Golabek A, Coates T, Kopczynski J, Gozdz S, Miettinen M. Detection of the BRAF V600E mutation in colon carcinoma: critical evaluation of the imunohistochemical approach. The American journal of surgical pathology. 2014 Sep 1;38(9):1235-41.
Zhao J, Liu P, Yu Y, Zhi J, Zheng X, Yu J, Gao M. Comparison of diagnostic methods for the detection of a BRAF mutation in papillary thyroid cancer. Oncology Letters. 2019 May 1;17(5):4661-6.
Zhu X, Luo Y, Bai Q, Lu Y, Lu Y, Wu L, Zhou X. Specific immunohistochemical detection of the BRAF V600E mutation in primary and metastatic papillary thyroid carcinoma. Experimental and Molecular Pathology. 2016 Feb 1;100(1):236- 41.
Martinuzzi C, Pastorino L, Andreotti V, Garuti A, Minuto M, Fiocca R, Bianchi-Scarrà G, Ghiorzo P, Grillo F, Mastracci L. A combination of immunohistochemistry and molecular approaches improves highly sensitive detection of BRAF mutations in papillary thyroid cancer. Endocrine. 2016 Sep;53(3):672-80.
Rashid FA, Tabassum S, Khan MS, Ansari HR, Asif M, Sheikh AK, Sameer Aga S. VE1 immunohistochemistry is an adjunct tool for detection of BRAFV600E mutation: Validation in thyroid cancer patients. Journal of Clinical Laboratory Analysis. 2021 Feb;35(2):e23628.
Choden S, Keelawat S, Jung CK, Bychkov A. An affordable immunohistochemical approach to estimate the prevalence of BRAF(V600E) in large cohort studies-establishing the baseline rate of BRAF mutation in an institutional series of papillary thyroid carcinoma from Thailand. Gland Surg. 2020;9(5):1867-77.
Escobar J, Guzmán GE, Urbano MA, Ballen LJ, Martínez V, Arrunategui A. Histological and Immunohistochemical Braf V600e Mutation Detection in Papillary Thyroid Carcinoma. Research Square. 2022;10(5):1-13
Kaliszewski K, Diakowska D, Nowak L, Wojtczak B, Rudnicki J. The age threshold of the 8th edition AJCC classification is useful for indicating patients with aggressive papillary thyroid cancer in clinical practice. BMC Cancer. 2020; 20(1):1166.
Abdullah MI, Junit SM, Ng KL, Jayapalan JJ, Karikalan B, Hashim OH. Papillary Thyroid Cancer: Genetic Alterations and Molecular Biomarker Investigations. Int J Med Sci. 2019;16(3):450-60.
Drake T. What Is the Ideal Age Cutoff for Papillary and Follicular Thyroid Cancer Staging, and Should the Cutoffs Be Different? Clinical Thyroidology. 2021;33(4):177-9.
Cho JS, Yoon JH, Park MH, Shin SH, Jegal YJ, Lee JS, Kim HK. Age and prognosis of papillary thyroid carcinoma: retrospective stratification into three groups. Journal of the Korean Surgical Society. 2012 Nov 1;83(5):259-66.
Szymonek M, Kowalik A, Kopczyński J, Gąsior-Perczak D, Pałyga I, Walczyk A, Gadawska-Juszczyk K, Płusa A, Mężyk R, Chrapek M, Góźdź S. Immunohistochemistry cannot replace DNA analysis for evaluation of BRAF V600E mutations in papillary thyroid carcinoma. Oncotarget. 2017 Sep 26;8(43):74897.
Girardi FM. Thyroid Carcinoma Pattern Presentation According to Age. Int Arch Otorhinolaryngol. 2017;21(1):38-41.
LeClair K, Bell KJL, Furuya-Kanamori L, Doi SA, Francis DO, Davies L. Evaluation of Gender Inequity in Thyroid Cancer Diagnosis: Differences by Sex in US Thyroid Cancer Incidence Compared With a Meta-analysis of Subclinical Thyroid Cancer Rates at Autopsy. JAMA Intern Med. 2021;181(10):1351- 8.
Jonklaas J, Nogueras-Gonzalez G, Munsell M, Litofsky D, Ain KB, Bigos ST, Brierley JD, Cooper DS, Haugen BR, Ladenson PW, Magner J. The impact of age and gender on papillary thyroid cancer survival. The Journal of Clinical Endocrinology & Metabolism. 2012 Jun 1;97(6):E878-87.
Rahbari R, Zhang L, Kebebew E. Thyroid cancer gender disparity. Future Oncology. 2010 Nov;6(11):1771-9.
Qiu T, Lu H, Guo L, Huang W, Ling Y, Shan L, Li W, Ying J, Lv N. Detection of BRAF mutation in Chinese tumor patients using a highly sensitive antibody immunohistochemistry assay. Scientific reports. 2015 Mar 18;5(1):1-4.
Zhang X, Wang L, Wang J, Zhao H, Wu J, Liu S, Zhang L, Li Y, Xing X. Immunohistochemistry is a feasible method to screen BRAF V600E mutation in colorectal and papillary thyroid carcinoma. Experimental and Molecular Pathology. 2018 Aug 1;105(1):153-9.
Shi SR, Gu J, Turrens J, Cote RJ, Taylor CR. Development of the antigen retrieval technique: philosophical and theoretical bases. Antigen retrieval techniques: immunohistochemistry and molecular morphology. 2000:17-40.
Anitha N, Malathy L. Immunohistochemistry-a brief note on technical protocols. European Journal of Molecular & Clinical Medicine. 2020 Dec 16;7(10):696-702.
Shi Z, Stack M. An Update on Immunohistochemistry in Translational Cancer Research. Cancer Translational Medicine. 2015;1(4):115.
Bussolati G, Leonardo E. Technical pitfalls potentially affecting diagnoses in immunohistochemistry. J Clin Pathol. 2008;61(11):1184-92.
Shi Z, Stack M. An Update on Immunohistochemistry in Translational Cancer Research. Cancer Translational Medicine. 2015;1(4): 115-22.
Parker KG, White MG, Cipriani NA. Comparison of Molecular Methods and BRAF Immunohistochemistry (VE1 Clone) for the Detection of BRAF V600E Mutation in Papillary Thyroid Carcinoma: A Meta-Analysis. Head Neck Pathol. 2020; 14(4):1067-79.
De Biase D, Cesari V, Visani M, Casadei GP, Cremonini N, Gandolfi G, Sancisi V, Ragazzi M, Pession A, Ciarrocchi A, Tallini G. High-sensitivity BRAF mutation analysis: BRAF V600E is acquired early during tumor development but is heterogeneously distributed in a subset of papillary thyroid carcinomas. The Journal of Clinical Endocrinology & Metabolism. 2014 Aug 1;99(8):E1530-8.
Kristiani E, Hardjolukito ES, Harahap AS, Makes B. BRAF V600E Immunoexpression in Papillary Thyroid Carcinoma and Its Association with Prognostic Factors and Histopathologic Variant. Medicinus. 2021;8(1):12-21.
Choden S, Keelawat S, Jung CK, Bychkov A. VE1 Immunohistochemistry Improves the Limit of Genotyping for Detecting BRAF(V600E) Mutation in Papillary Thyroid Cancer. Cancers (Basel). 2020;12(3):596.
Fonseca D, Murthy SS, Tagore R, Rao V, Rao CS, Raju KV, Nemade HK, Challa S. BRAF status in the variants of papillary thyroid carcinoma. International Journal of Head and Neck Pathology. 2018 Jul 1;1(2):41.
Zhang B, Xu CW, Wu YF, Man QH, Song YY, Wang JJ, Wang HT, Wang HY, Li XB, Zhang H, Ye T. Diagnostic significance of the BRAF V600E mutation in conventional papillary thyroid carcinomas. Int J Clin Exp Med. 2016 Jan 1;9(5):8296-303.
Jung YY, Yoo JH, Park ES, Kim MK, Lee TJ, Cho BY, Chung YJ, Kang KH, Ahn HY, Kim HS. Clinicopathologic correlations of the BRAFV600E mutation, BRAF V600E immunohistochemistry, and BRAF RNA in situ hybridization in papillary thyroid carcinoma. Pathology-Research and Practice. 2015 Feb 1;211(2):162-70.
Zhang Y, Liu L, Liu Y, Cao N, Wang L, Xing C. Clinical significance of immunohistochemistry to detect BRAF V600E mutant protein in thyroid tissues. Medicine. 2021 Apr 23;100(16).
Kauffmann RM, Hamner JB, Ituarte PHG, Yim JH. Age greater than 60 years portends a worse prognosis in patients with papillary thyroid cancer: should there be three age categories for staging? BMC Cancer. 2018;18(1):316.
Li C, Lee KC, Schneider EB, Zeiger MA. BRAF V600E mutation and its association with clinicopathological features of papillary thyroid cancer: a meta-analysis. J Clin Endocrinol Metab. 2012;97(12):4559-70.
Koperek O, Kornauth C, Capper D, Berghoff AS, Asari R, Niederle B, et al. Immunohistochemical detection of the BRAF V600E-mutated protein in papillary thyroid carcinoma. The American Journal of Surgical Pathology. 2012;36 (6):844-50.
Sun J, Zhang J, Lu J, Gao J, Lu T, Ren X, Duan H, Liang Z. Immunohistochemistry is highly sensitive and specific for detecting the BRAF V600E mutation in papillary thyroid carcinoma. International journal of clinical and experimental pathology. 2015;8(11):15072.
Zagzag J, Pollack A, Dultz L, Dhar S, Ogilvie JB, Heller KS, Deng FM, Patel KN. Clinical utility of immunohistochemistry for the detection of the BRAF v600e mutation in papillary thyroid carcinoma. Surgery. 2013 Dec 1;154(6):1199-205.
Barreno LR, Mello JB, Barros-Filho MC, Francisco AL, Chulam TC, Pinto CA, Gonçalves-Filho J, Kowalski LP. Characterization of BRAF mutation in patients older than 45 years with well-differentiated thyroid carcinoma. Brazilian Journal of Otorhinolaryngology. 2022 Aug 15;88:523-8.
McKelvie PA, Chan F, Yu Y, Waring P, Gresshoff I, Farrell S, Williams RA. The prognostic significance of the BRAFV600E mutation in papillary thyroid carcinoma detected by mutation-specific immunohistochemistry. Pathology. 2013 Dec 1;45(7):637-44.
Ilie MI, Lassalle S, Long-Mira E, Bonnetaud C, Bordone O, Lespinet V, Lamy A, Sabourin JC, Haudebourg J, Butori C, Guevara N. Diagnostic value of immunohistochemistry for the detection of the BRAFV600E mutation in papillary thyroid carcinoma: comparative analysis with three DNA-based assays. Thyroid. 2014 May 1;24(5):858-66.
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