BRAF Mutational Analysis in Papillary Carcinomas With Mixed Follicular and Papillary Growth Patterns

Abstract

Current literature suggests that up to 60% of papillary carcinomas have mutations in the BRAF gene. However, follicular variant of papillary carcinoma has a much lower frequency of mutation. Tumors with mixed patterns of growth, including distinctive area of follicular and papillary growth, have not been well studied for the presence of the BRAF gene mutation. Cases of papillary carcinoma were identified with well-defined conventional papillary growth pattern, alongside other areas with follicular growth pattern. The tumor stage was obtained, along with demographic information. The different growth pattern areas were separately microdissected and DNA was extracted from the resulting tissue fragments. Polymerase chain reaction was performed for an amplicon in BRAF exon 15 that includes the mutation site. Cycle sequencing was performed using the BigDye Terminator kit and analysis was performed on an ABI automated sequencer (Applied Biosystems). The forward and reverse sequences were analyzed for point mutations. DNA extraction and polymerase chain reaction amplification was successful in all cases. Seventy percent of the tumors were positive for the BRAF mutation. In comparing the follicular and the papillary growth patterned areas, the BRAF mutation was concordant in all cases. Four cases had an additional separate focus of microscopic papillary carcinomas. These were negative for the BRAF mutation. Papillary carcinomas of the thyroid with papillary growth and areas of follicular growth have a high frequency of BRAF mutations. The BRAF mutational profile is identical in the follicular areas and in the conventional papillary growth areas. These molecular data support the common diagnostic decision that a tumor with any amount of conventional papillary growth should be designated as a conventional papillary carcinoma, regardless of the presence of follicular growth pattern areas.