Telomerase reverse transcriptase (TERT) promoter mutations occur at a high frequency in melanoma, but the functional consequences of these mutations in melanoma remain to be clarified. In a large study of melanoma samples by The Cancer Genome Atlas network, mRNA expression analysis using RNA sequencing data showed that only TERT promoter mutations at C228T were associated with high TERT expression levels. The effect of hotspot C250 or C242T/C243T tandem mutations on telomerase expression has not yet been determined. To assess telomerase expression levels in melanomas harboring C250 or C242T/C243T mutations, we used real-time quantitative reverse transcriptase polymerase chain reaction (RT-qPCR) in a sample set of pediatric melanocytic tumors for which the methylation status and the mutational profile of the TERT promoter were known. We previously showed that in a subset of melanoma, TERT expression is mediated epigenetically by promoter hypermethylation rather than by promoter point mutations. Formalin-fixed paraffin-embedded (FFPE) tissues from 10 metastatic melanomas, of which 8 harbored a hotspot TERT promoter mutation (5 C250T; 2 C228; 1 C242T/C243T) and 2 carried a hypermethylated wild-type TERT promoter were examined. For controls, 9 atypical Spitz tumors with a wild-type unmethylated TERT promoter were also studied. Total RNA was isolated from FFPE tumor tissues and converted to cDNA. TERT expression levels were determined by RT-qPCR by using the TaqMan® Gene Expression Assay and normalized with GAPDH as the endogenous control. In the atypical Spitz tumor samples, TERT mRNA expression was undetectable or negligible. In contrast, TERT expression in all melanoma samples was elevated, ranging from 3- to 71-fold (median, 25-fold) when compared to an atypical Spitz tumor as a reference. Our data support that in melanoma, TERT promoter hotspot C250 and C242T/C243T mutations, similar to C228T mutations, correlate with TERT overexpression and that C250 and C242T/C243T mutations likely contribute biologically to tumorigenesis in melanoma.

Citation Format: Seungjae Lee, Patricia Opresko, Alberto Pappo, John Kirkwood, Armita Bahrami. Association of TERT promoter mutations with telomerase expression in melanoma. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 2730.