Yonsei Med J.  2015 May;56(3):634-640. 10.3349/ymj.2015.56.3.634.

Detection of Plasma BRAF(V600E) Mutation Is Associated with Lung Metastasis in Papillary Thyroid Carcinomas

• Kim BH
• Kim IJ
• Lee BJ
• Lee JC
• Kim IS
• Kim SJ
• Kim WJ
• Jeon YK
• Kim SS
• Kim YK

• KMID: 2450335
• DOI: http://doi.org/10.3349/ymj.2015.56.3.634

Abstract

PURPOSE
The BRAF(V600E) mutation represents a novel indicator of the progression and aggressiveness of papillary thyroid carcinoma (PTC). The purpose of this study was to determine the clinical significance of free circulating mutant BRAF(V600E) in predicting the advanced disease of PTC.
MATERIALS AND METHODS
Seventy seven matched tumor and plasma samples obtained from patients with both benign and PTC were analyzed for BRAF(V600E) mutation using a peptide nucleic acid (PNA) clamp real-time polymerase chain reaction (PCR).
RESULTS
The BRAF(V600E) mutation was absent in tumor DNA samples obtained from patients with benign follicular adenomas or adenomatous goiter. In contrast, 49 of 72 (68.1%) PTC tumors were positive for the BRAF(V600E) mutation. Among them, 3 (6.1%) patients with PTC were positive for BRAF(V600E) mutation in plasma and tumor. However, all 3 patients (100%) had lateral lymph node and lung metastasis.
CONCLUSION
These findings suggest that the BRAF(V600E) mutation can be detected using a PNA clamp real-time PCR in the blood of PTC patients with lung metastasis. Future studies are warranted to determine clinical significance of serum BRAF(V600E) mutation in large prospective studies.

Keyword

BRAF; papillary thyroid carcinoma; plasma; DNA; biomarker

MeSH Terms

Adenocarcinoma, Papillary/*genetics/secondary
Adult
Aged
Carcinoma/*genetics/pathology
DNA Mutational Analysis
DNA, Neoplasm/*genetics
Female
Humans
Lung Neoplasms/*genetics
Lymph Nodes/pathology
Lymphatic Metastasis
Male
Middle Aged
Mutation
Neoplasm Invasiveness
Neoplasm Staging
*Peptide Nucleic Acids
Prospective Studies
Proto-Oncogene Proteins B-raf/*genetics
Real-Time Polymerase Chain Reaction
Thyroid Neoplasms/*genetics/pathology
DNA, Neoplasm
Peptide Nucleic Acids
Proto-Oncogene Proteins B-raf

Figure

• 

  Fig. 1 PNA Clamp™ standard curves in tumor DNA samples and plasma DNA sample. The detection signal was obtained by intercalation of SYBR green fluorescent dye of real-time PCR. A PNA/DNA hybrid with a single base-pair mismatch did not suppress annealing of the PCR primer (A) and amplification of mutant alleles (B). PNA, peptide nucleic acid; PCR, polemerase chain reaction; RFU, relative fluorescence units.

• 

  Fig. 2 Using the direct sequencing of DNA, representative sequence chromatographs from BRAF axon 15 showing wild type (A) and mutation (B) in papillary thyroid carcinomas. Arrows indicate mutations.

Reference

1. Mazzaferri EL. Long-term outcome of patients with differentiated thyroid carcinoma: effect of therapy. Endocr Pract. 2000; 6:469–476.

2. Grebe SK, Hay ID. Thyroid cancer nodal metastases: biologic significance and therapeutic considerations. Surg Oncol Clin N Am. 1996; 5:43–63.

3. Machens A, Hinze R, Thomusch O, Dralle H. Pattern of nodal metastasis for primary and reoperative thyroid cancer. World J Surg. 2002; 26:22–28.

4. Stulak JM, Grant CS, Farley DR, Thompson GB, van Heerden JA, Hay ID, et al. Value of preoperative ultrasonography in the surgical management of initial and reoperative papillary thyroid cancer. Arch Surg. 2006; 141:489–494.

5. Shaha AR, Ferlito A, Rinaldo A. Distant metastases from thyroid and parathyroid cancer. ORL J Otorhinolaryngol Relat Spec. 2001; 63:243–249.

6. Benbassat CA, Mechlis-Frish S, Hirsch D. Clinicopathological characteristics and long-term outcome in patients with distant metastases from differentiated thyroid cancer. World J Surg. 2006; 30:1088–1095.

7. Liu D, Liu Z, Condouris S, Xing M. BRAF V600E maintains proliferation, transformation, and tumorigenicity of BRAF-mutant papillary thyroid cancer cells. J Clin Endocrinol Metab. 2007; 92:2264–2271.

8. Cradic KW, Milosevic D, Rosenberg AM, Erickson LA, McIver B, Grebe SK. Mutant BRAF(T1799A) can be detected in the blood of papillary thyroid carcinoma patients and correlates with disease status. J Clin Endocrinol Metab. 2009; 94:5001–5009.

9. Chuang TC, Chuang AY, Poeta L, Koch WM, Califano JA, Tufano RP. Detectable BRAF mutation in serum DNA samples from patients with papillary thyroid carcinomas. Head Neck. 2010; 32:229–234.

10. Kim KH, Kang DW, Kim SH, Seong IO, Kang DY. Mutations of the BRAF gene in papillary thyroid carcinoma in a Korean population. Yonsei Med J. 2004; 45:818–821.

11. Kim TY, Kim WB, Rhee YS, Song JY, Kim JM, Gong G, et al. The BRAF mutation is useful for prediction of clinical recurrence in low-risk patients with conventional papillary thyroid carcinoma. Clin Endocrinol (Oxf). 2006; 65:364–368.

12. Jeong D, Jeong Y, Park JH, Han SW, Kim SY, Kim YJ, et al. BRAF (V600E) mutation analysis in papillary thyroid carcinomas by peptide nucleic acid clamp real-time PCR. Ann Surg Oncol. 2013; 20:759–766.

13. Kwon MJ, Lee SE, Kang SY, Choi YL. Frequency of KRAS, BRAF, and PIK3CA mutations in advanced colorectal cancers: comparison of peptide nucleic acid-mediated PCR clamping and direct sequencing in formalin-fixed, paraffin-embedded tissue. Pathol Res Pract. 2011; 207:762–768.

14. Zaydfudim V, Feurer ID, Griffin MR, Phay JE. The impact of lymph node involvement on survival in patients with papillary and follicular thyroid carcinoma. Surgery. 2008; 144:1070–1077.

15. Mazzaferri EL. Management of a solitary thyroid nodule. N Engl J Med. 1993; 328:553–559.

16. Riesco-Eizaguirre G, Gutiérrez-Martínez P, García-Cabezas MA, Nistal M, Santisteban P. The oncogene BRAF V600E is associated with a high risk of recurrence and less differentiated papillary thyroid carcinoma due to the impairment of Na+/I- targeting to the membrane. Endocr Relat Cancer. 2006; 13:257–269.

17. Elisei R, Ugolini C, Viola D, Lupi C, Biagini A, Giannini R, et al. BRAF(V600E) mutation and outcome of patients with papillary thyroid carcinoma: a 15-year median follow-up study. J Clin Endocrinol Metab. 2008; 93:3943–3949.

18. 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; 92:4085–4090.

19. Lee JH, Lee ES, Kim YS. Clinicopathologic significance of BRAF V600E mutation in papillary carcinomas of the thyroid: a meta-analysis. Cancer. 2007; 110:38–46.

20. 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; 246:466–470.

21. Trovisco V, Couto JP, Cameselle-Teijeiro J, de Castro IV, Fonseca E, Soares P, et al. Acquisition of BRAF gene mutations is not a requirement for nodal metastasis of papillary thyroid carcinoma. Clin Endocrinol (Oxf). 2008; 69:683–685.

22. Xing M. BRAF mutation in papillary thyroid cancer: pathogenic role, molecular bases, and clinical implications. Endocr Rev. 2007; 28:742–762.

23. Guan H, Ji M, Bao R, Yu H, Wang Y, Hou P, et al. Association of high iodine intake with the T1799A BRAF mutation in papillary thyroid cancer. J Clin Endocrinol Metab. 2009; 94:1612–1617.

24. Kim JY, Moon SJ, Kim KR, Sohn CY, Oh JJ. Dietary iodine intake and urinary iodine excretion in normal Korean adults. Yonsei Med J. 1998; 39:355–362.

25. Kang SH, Pyo JY, Yang SW, Hong SW. Detection of BRAF V600E mutation with thyroid tissue using pyrosequencing: comparison with PNA-clamping and real-time PCR. Am J Clin Pathol. 2013; 139:759–764.

26. Kim TY, Kim WB, Song JY, Rhee YS, Gong G, Cho YM, et al. The BRAF mutation is not associated with poor prognostic factors in Korean patients with conventional papillary thyroid microcarcinoma. Clin Endocrinol (Oxf). 2005; 63:588–593.

27. Rha SY, Lee JC, Kwon KH, Lee HJ, Kim KS, Jo YS, et al. The relationship between the BRAF mutations in thyroid papillary carcinomas and the prognostic factors. J Korean Soc Endocrinol. 2005; 20:224–229.

28. Trovisco V, Soares P, Preto A, de Castro IV, Lima J, Castro P, et al. Type and prevalence of BRAF mutations are closely associated with papillary thyroid carcinoma histotype and patients' age but not with tumour aggressiveness. Virchows Arch. 2005; 446:589–595.