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Significance of Allelic Percentage of BRAF c.1799T > A (V600E) Mutation in Papillary Thyroid Carcinoma

  • Translational Research and Biomarkers
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Abstract

Background

Somatic BRAF mutation is frequently observed in papillary thyroid carcinoma (PTC). Recent evidence suggests that PTCs are heterogeneous tumors containing a subclonal or oligoclonal occurrence of BRAF mutation. Conflicting results have been reported concerning the prognostic significance of the mutant allele frequency. Our present aim was to investigate the association between the percentage of BRAF c.1799T > A (p.Val600Glu) alleles and clinicopathological parameters in PTC.

Methods

Genomic DNA was extracted from fresh-frozen specimens obtained from 50 PTC patients undergoing total thyroidectomy. The BRAF mutation status was determined by Sanger sequencing. The percentage of mutant BRAF alleles was quantified by mass spectrometric genotyping, pyrosequencing, and competitive allele-specific TaqMan PCR (castPCR).

Results

Positive rate of BRAF mutation was 72 % by Sanger sequencing, 82 % by mass spectrometric genotying, and 84 % by pyrosequencing or castPCR. The average percentage of mutant BRAF alleles was 22.5, 31, and 30.7 %, respectively. There was a good correlation among three quantification methods (Spearman’s rho = 0.87–0.97; p < 0.0001). The mutant allele frequency was significantly correlated with tumor size (rho = 0.47–0.52; p < 0.01) and extrathyroidal invasion. The frequency showed no difference in pathological lymph node metastasis.

Conclusions

The percentage of mutant BRAF alleles is positively associated with tumor burden and extrathyroidal invasion in PTC. Relatively good correlations exist among mass spectrometric genotyping, pyrosequencing, and castPCR in quantification of mutant BRAF allele frequency.

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Acknowledgment

The authors would like to thank the Translational Core Laboratories of National Translational Medicine and Clinical Trial Resource Center for technical assistance in MALDI-TOF MS experiments. This work was supported by the National Science Council of Taiwan (100-2314-B-195-001-MY3) and Mackay Memorial Hospital (MMH-10206 and MMH-E-102-10).

Disclosures

Shih-Ping Cheng, Yi-Chiung Hsu, Chien-Liang Liu, Tsang-Pai Liu, Ming-Nan Chien, Tao-Yeuan Wang, and Jie-Jen Lee have nothing to disclose.

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Authors and Affiliations

  1. Mackay Junior College of Medicine, Nursing, and Management, Taipei, Taiwan

    Shih-Ping Cheng MD, PhD, Chien-Liang Liu MD, Tsang-Pai Liu MD, Ming-Nan Chien MD, Tao-Yeuan Wang MD & Jie-Jen Lee MD, PhD

  2. Department of Surgery, Mackay Medical College and Mackay Memorial Hospital, Taipei, Taiwan

    Shih-Ping Cheng MD, PhD, Chien-Liang Liu MD, Tsang-Pai Liu MD & Jie-Jen Lee MD, PhD

  3. Division of Endocrinology and Metabolism, Department of Medicine, Mackay Medical College and Mackay Memorial Hospital, Taipei, Taiwan

    Ming-Nan Chien MD

  4. Department of Pathology, Mackay Medical College and Mackay Memorial Hospital, Taipei, Taiwan

    Tao-Yeuan Wang MD

  5. Institute of Statistical Science, Academia Sinica, Taipei, Taiwan

    Yi-Chiung Hsu PhD

  6. Graduate Institute of Medical Sciences and Department of Pharmacology, Taipei Medical University, Taipei, Taiwan

    Shih-Ping Cheng MD, PhD & Jie-Jen Lee MD, PhD

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  1. Shih-Ping Cheng MD, PhD
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Cheng, SP., Hsu, YC., Liu, CL. et al. Significance of Allelic Percentage of BRAF c.1799T > A (V600E) Mutation in Papillary Thyroid Carcinoma. Ann Surg Oncol 21 (Suppl 4), 619–626 (2014). https://doi.org/10.1245/s10434-014-3723-5

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  • DOI: https://doi.org/10.1245/s10434-014-3723-5

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