.: ORİJİNAL ARAŞTIRMA
Analysis of BRAFV600E, KRAS Codon 12/13 and Novel Mutations in Patients with Thyroid Cancer in Blacksea Region of Turkey
Karadeniz Bölgesinde Tiroid Kanserli Hastalarda BRAFV600E, KRAS Kodon 12/13 ve Yeni Mutasyonların Analizi
Turkiye Klinikleri J Med Sci. 2021;41(1):80-5
doi: 10.5336/medsci.2020-76105
Makale Dili: EN
Tam Metin
ABSTRACT
Objective: The incidence of thyroid cancer in many regions of the world has been steadily increasing over the past decades. Point mutations in the B-Raf (BRAF) and RAS proto-oncogenes are crucial in the molecular pathogenesis of thyroid cancers. In this study, it was aimed to investigate the BRAFV600E and KRAS codon 12/13 mutations in thyroid cancer patients of the Turkish population. Material and Methods: In this study, totally 32 cases with thyroid cancer were investigated. Ten of the paraffin-embedded thyroid cancer tissues and 22 of peripheral blood samples from patients were included in the study. Genomic DNA was extracted from the paraffin-embedded sections and peripheral blood samples, and then polymerase chain reaction and DNA sequencing were performed for mutation analysis of BRAF and KRAS genes. Results: BRAFV600E mutations were found in 13 (40.6%) patients, and 19 patients (59.4%) had wild-type profiles. G12N, G12V, and G13V mutations that are frequently seen in KRAS were not observed, but four new mutations (K16E, L19G, E31D, and M1I) were found in the current cohort. Conclusion: BRAFV600E mutation was detected in patients with thyroid cancer, and four new mutations, which are in the GTP binding regions of the protein, were detected in the KRAS gene. These new mutations might be related to papillary thyroid cancer.
Keywords: Proto-oncogene proteins B-Raf; genes, Ras; thyroid cancer, papillary; mutation
Objective: The incidence of thyroid cancer in many regions of the world has been steadily increasing over the past decades. Point mutations in the B-Raf (BRAF) and RAS proto-oncogenes are crucial in the molecular pathogenesis of thyroid cancers. In this study, it was aimed to investigate the BRAFV600E and KRAS codon 12/13 mutations in thyroid cancer patients of the Turkish population. Material and Methods: In this study, totally 32 cases with thyroid cancer were investigated. Ten of the paraffin-embedded thyroid cancer tissues and 22 of peripheral blood samples from patients were included in the study. Genomic DNA was extracted from the paraffin-embedded sections and peripheral blood samples, and then polymerase chain reaction and DNA sequencing were performed for mutation analysis of BRAF and KRAS genes. Results: BRAFV600E mutations were found in 13 (40.6%) patients, and 19 patients (59.4%) had wild-type profiles. G12N, G12V, and G13V mutations that are frequently seen in KRAS were not observed, but four new mutations (K16E, L19G, E31D, and M1I) were found in the current cohort. Conclusion: BRAFV600E mutation was detected in patients with thyroid cancer, and four new mutations, which are in the GTP binding regions of the protein, were detected in the KRAS gene. These new mutations might be related to papillary thyroid cancer.
Keywords: Proto-oncogene proteins B-Raf; genes, Ras; thyroid cancer, papillary; mutation
ÖZET
Amaç: Dünyanın pek çok bölgesinde tiroid kanseri görülme sıklığı, son yıllarda sürekli olarak artmaktadır. B-Raf (BRAF) ve RAS proto-onkogenlerindeki nokta mutasyonları, tiroid kanserlerinin moleküler patogenezinde çok önemlidir. Bu çalışmada, Türk toplumundaki tiroid kanseri hastalarında BRAFV600E ve KRAS kodon 12/13 mutasyonlarının araştırılması amaçlandı. Gereç ve Yöntemler: Çalışmada, toplam 32 tiroid kanseri vakası incelendi. Parafine gömülü tiroid kanser dokularından 10'u ve hastalardan alınan periferik kan örneklerinden 22'si çalışmaya dâhil edildi. Parafine gömülü kesitlerden ve periferik kan örneklerinden genomik DNA ekstrakte edildikten sonra BRAF ve KRAS genlerinin mutasyon analizi için polimeraz zincir reaksiyonu ve DNA dizi analizi yapıldı. Bulgular: BRAFV600E mutasyonları 13 (%40,6) hastada bulundu ve 19 (%59,4) hasta ise yabanıl tip profile sahipti. KRAS'ta sıklıkla görülen G12N, G12V ve G13V mutasyonları bu kohortta gözlenmedi, ancak 4 yeni mutasyon (K16E, L19G, E31D ve M1I) bulundu. Sonuç: Tiroid kanserli hastalarda BRAFV600E mutasyonu ve KRAS geninde proteinin GTP bağlanma bölgelerinde meydana gelen 4 yeni mutasyon tespit edildi. Bu yeni mutasyonlar papiller tiroid kanseriyle ilişkili olabilir.
Anahtar Kelimeler: Proto-onkogen proteinleri B-Raf; genler, Ras; tiroid kanser, papiller; mutasyon
Amaç: Dünyanın pek çok bölgesinde tiroid kanseri görülme sıklığı, son yıllarda sürekli olarak artmaktadır. B-Raf (BRAF) ve RAS proto-onkogenlerindeki nokta mutasyonları, tiroid kanserlerinin moleküler patogenezinde çok önemlidir. Bu çalışmada, Türk toplumundaki tiroid kanseri hastalarında BRAFV600E ve KRAS kodon 12/13 mutasyonlarının araştırılması amaçlandı. Gereç ve Yöntemler: Çalışmada, toplam 32 tiroid kanseri vakası incelendi. Parafine gömülü tiroid kanser dokularından 10'u ve hastalardan alınan periferik kan örneklerinden 22'si çalışmaya dâhil edildi. Parafine gömülü kesitlerden ve periferik kan örneklerinden genomik DNA ekstrakte edildikten sonra BRAF ve KRAS genlerinin mutasyon analizi için polimeraz zincir reaksiyonu ve DNA dizi analizi yapıldı. Bulgular: BRAFV600E mutasyonları 13 (%40,6) hastada bulundu ve 19 (%59,4) hasta ise yabanıl tip profile sahipti. KRAS'ta sıklıkla görülen G12N, G12V ve G13V mutasyonları bu kohortta gözlenmedi, ancak 4 yeni mutasyon (K16E, L19G, E31D ve M1I) bulundu. Sonuç: Tiroid kanserli hastalarda BRAFV600E mutasyonu ve KRAS geninde proteinin GTP bağlanma bölgelerinde meydana gelen 4 yeni mutasyon tespit edildi. Bu yeni mutasyonlar papiller tiroid kanseriyle ilişkili olabilir.
Anahtar Kelimeler: Proto-onkogen proteinleri B-Raf; genler, Ras; tiroid kanser, papiller; mutasyon
REFERANSLAR:
- Nikiforov YE, Nikiforova MN. Molecular genetics and diagnosis of thyroid cancer. Nat Rev Endocrinol. 2011;7(10):569-80.[Crossref] [PubMed]
- Albores-Saavedra J, Henson DE, Glazer E, Schwartz AM. Changing patterns in the incidence and survival of thyroid cancer with follicular phenotype--papillary, follicular, and anaplastic: a morphological and epidemiological study. Endocr Pathol. 2007;18(1):1-7.[Crossref] [PubMed]
- Ron E, Lubin JH, Shore RE, Mabuchi K, Modan B, Pottern LM, et al. Thyroid cancer after exposure to external radiation: a pooled analysis of seven studies. Radiat Res. 1995;141(3):259-77.[Crossref] [PubMed]
- Enewold L, Zhu K, Ron E, Marrogi AJ, Stojadinovic A, Peoples GE, et al. Rising thyroid cancer incidence in the United States by demographic and tumor characteristics, 1980-2005. Cancer Epidemiol Biomarkers Prev. 2009;18(3):784-91.[Crossref] [PubMed] [PMC]
- Kimura ET, Nikiforova MN, Zhu Z, Knauf JA, Nikiforov YE, Fagin JA. High prevalence of BRAF mutations in thyroid cancer: genetic evidence for constitutive activation of the RET/PTC-RAS-BRAF signaling pathway in papillary thyroid carcinoma. Cancer Res. 2003;63(7):1454-7.[PubMed]
- Soares P, Trovisco V, Rocha AS, Lima J, Castro P, Preto A, et al. BRAF mutations and RET/PTC rearrangements are alternative events in the etiopathogenesis of PTC. Oncogene. 2003;22(29):4578-80.[Crossref] [PubMed]
- Nikiforov YE, Nikiforova MN. Molecular genetics and diagnosis of thyroid cancer. Nat Rev Endocrinol. 2011;7(10):569-80.[Crossref] [PubMed]
- Tufano RP, Teixeira GV, Bishop J, Carson KA, Xing M. BRAF mutation in papillary thyroid cancer and its value in tailoring initial treatment: a systematic review and meta-analysis. Medicine (Baltimore). 2012;91(5):274-86.[Crossref] [PubMed]
- Sithanandam G, Druck T, Cannizzaro LA, Leuzzi G, Huebner K, Rapp UR. B-raf and a B-raf pseudogene are located on 7q in man. Oncogene. 1992;7(4):795-9.[PubMed]
- Cohen Y, Xing M, Mambo E, Guo Z, Wu G, Trink B, et al. BRAF mutation in papillary thyroid carcinoma. J Natl Cancer Inst. 2003;95(8):625-7.[Crossref] [PubMed]
- Xing M. BRAF mutation in thyroid cancer. Endocr Relat Cancer. 2005;12(2):245-62.[Crossref] [PubMed]
- Kranenburg O. The KRAS oncogene: past, present, and future. Biochim Biophys Acta. 2005 Nov 25;1756(2):81-2.[Crossref] [PubMed]
- Ozen F, Ozdemir S, Zemheri E, Hacimuto G, Silan F, Ozdemir O. The proto-oncogene KRAS and BRAF profiles and some clinical characteristics in colorectal cancer in the Turkish population. Genet Test Mol Biomarkers. 2013;17(2):135-9.[Crossref] [PubMed]
- Okayama N, Nishioka M, Hazama S, Sakai K, Suehiro Y, Maekawa M, et al. The importance of evaluation of DNA amplificability in KRAS mutation testing with dideoxy sequencing using formalin-fixed and paraffin-embedded colorectal cancer tissues. Jpn J Clin Oncol. 2011;41(2):165-71.[Crossref] [PubMed]
- Edge S, Byrd DR, Compton CC, Fritz AG, Greene F, Trotti A. AJCC Cancer Staging Handbook. 7th ed. New York/USA: Springer; 2010.[Link]
- Rako I, Jakic-Razumovic J, Katalinic D, Sertic J, Plestina S. Mutation pattern of KRAS and BRAF oncogenes in colorectal cancer patients. Neoplasma. 2012;59(4):376-83.[Crossref] [PubMed]
- Stanojevic B, Dzodic R, Saenko V, Milovanovic Z, Pupic G, Zivkovic O, et al. Mutational and clinico-pathological analysis of papillary thyroid carcinoma in Serbia. Endocr J. 2011;58(5):381-93.[Crossref] [PubMed]
- Trovisco V, Soares P, Sobrinho-Simões M. B-RAF mutations in the etiopathogenesis, diagnosis, and prognosis of thyroid carcinomas. Hum Pathol. 2006;37(7):781-6.[Crossref] [PubMed]
- Xu X, Quiros RM, Gattuso P, Ain KB, Prinz RA. High prevalence of BRAF gene mutation in papillary thyroid carcinomas and thyroid tumor cell lines. Cancer Res. 2003;63(15):4561-7.[PubMed]
- Lupi C, Giannini R, Ugolini C, Proietti A, Berti P, Minuto M, et al. Association of BRAF V600E mutation with poor clinicopathological outcomes in 500 consecutive cases of papillary thyroid carcinoma. J Clin Endocrinol Metab. 2007;92(11):4085-90.[Crossref] [PubMed]
- Ito Y, Yoshida H, Maruo R, Morita S, Takano T, Hirokawa M, et al. BRAF mutation in papillary thyroid carcinoma in a Japanese population: its lack of correlation with high-risk clinicopathological features and disease-free survival of patients. Endocr J. 2009;56(1):89-97.[Crossref] [PubMed]
- Kurtulmus N, Duren M, Ince U, Cengiz Yakicier M, Peker O, Aydın O, et al. BRAFV600E mutation in Turkish patients with papillary thyroid cancer: strong correlation with indicators of tumor aggressiveness. Endocrine. 2012;42(2):404-10.[Crossref] [PubMed]
- Kebebew E, Weng J, Bauer J, Ranvier G, Clark OH, Duh QY, et al. The prevalence and prognostic value of BRAF mutation in thyroid cancer. Ann Surg. 2007;246(3):466-70; discussion 470-1.[Crossref] [PubMed] [PMC]
- Czarniecka A, Rusinek D, Stobiecka E, Krajewska J, Kowal M, Kropińska A, et al. Occurrence of BRAF mutations in a Polish cohort of PTC patients - preliminary results. Endokrynol Pol. 2010;61(5):462-6.[PubMed]
- Fugazzola L, Puxeddu E, Avenia N, Romei C, Cirello V, Cavaliere A, et al. Correlation between B-RAFV600E mutation and clinico-pathologic parameters in papillary thyroid carcinoma: data from a multicentric Italian study and review of the literature. Endocr Relat Cancer. 2006;13(2):455-64.[Crossref] [PubMed]
- Jung CK, Little MP, Lubin JH, Brenner AV, Wells SA Jr, Sigurdson AJ, et al. The increase in thyroid cancer incidence during the last four decades is accompanied by a high frequency of BRAF mutations and a sharp increase in RAS mutations. J Clin Endocrinol Metab. 2014;99(2):E276-85.[Crossref] [PubMed] [PMC]
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