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RT-PCR detection of Tyrosinase, gp100, MART1/Melan-A, and TRP-2 gene transcripts in peripheral blood of melanoma patients

Aim. Pilot study with the aim to detect the expression of genes encoding tyrosinase, gp100, MART-1/Melan A and tyrosinase-related protein-2 (TRP-2) in peripheral blood of melanoma patients by reverse transcription - polymerase chain reaction (RT-PCR). Methods. 19 peripheral blood samples from 17 melanoma patients, majority of them with clinically metastatic disease were assayed. From each patient 7 to 8 ml of peripheral blood was collected into vacutainer cell preparation tube (CPT ; Becton Dickinson). Mononuclear cells were collected, total cellular RNA was extracted, reverse transcribed (cDNA) and by PCR assayed for the expression for melanocyte-associated transcripts of tyrosinase, gp100, MART1/Melan-A and TRP-2. Results. Gp100 gene expression was detected in 13 out of 19 samples, TRP-2 gene was detected in 4 samples which were also gp100 gene positive. Expression of tyrosinase and Melan-A/MART-1gene could not be detected. Expression of gp100 and of TRP-2 gene was detected in patients having recurrent and/or metastatic disease at the time of testing. Conclusions. Obtained results support the usage of RT-PCR assay in the indirect detection of melanoma cells in peripheral blood of melanoma patients. Since the transcripts for the tyrosinase gene and MART-1/Melan A gene were not detected additional optimization experiments of RT-PCR assay are required.

melanoma; peripheral blood; RT-PCR; tyrosinase; gp100; MART-1/Melan A; TRP-2

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