ABSTRACT
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: Nineteen peripheral blood samples were obtained from 17 melanoma patients. When tested, 15 of them presented with clinically detectable metastatic disease. Samples of peripheral blood (7 mL) were collected from each patient into vacutainer cell preparation tubes. Mononuclear cells were isolated, total cellular RNA extracted, and then used as a template for reverse transcription to complementary DNA (cDNA). The cDNA was thereafter assayed by PCR for the expression of melanocyte-associated transcripts of tyrosinase, gp100, MART1/Melan-A, and TRP-2 genes. RESULTS: Gp100 gene expression was detected in 13 out of 19 samples. In 4 of them, TRP-2 gene expression was also detectable. Expression of tyrosinase and Melan-A/MART-1 genes could not be observed. Interestingly, gp100 and TRP-2 gene transcripts were detected in patients having recurrent and/or metastatic disease at the time of testing. CONCLUSION: The results we obtained support the use of RT-PCR assay for indirect detection of melanoma cells in peripheral blood of melanoma patients. As the transcripts for the tyrosinase gene and MART-1/Melan A gene were not detected, additional optimization experiments of RT-PCR assay are required.
Subject(s)
Melanoma/genetics , Reverse Transcriptase Polymerase Chain Reaction , Skin Neoplasms/genetics , Adult , Aged , Epitopes/blood , Epitopes/genetics , Female , Gene Expression , Humans , Intramolecular Oxidoreductases/blood , Intramolecular Oxidoreductases/genetics , Male , Membrane Glycoproteins/blood , Membrane Glycoproteins/genetics , Middle Aged , Neoplasm Proteins/blood , Neoplasm Proteins/genetics , Sensitivity and Specificity , gp100 Melanoma AntigenABSTRACT
In the Department of Plastic Surgery of "Dubrava" University Hospital from 1993 to 1999 four patients were treated for radiation induced sarcoma. All of the patients were formerly operated for breast cancer and irradiated postoperatively. The mean time span between radiotherapy and development of sarcoma was 4.75 years. Four patients were treated with wide excision and immediate reconstruction with local flaps. One of the patients had fibrosarcoma, two patients had lymphangiosarcoma, and one patient had osteosarcoma. Two patients died within two years. Radiation induced sarcomas are therapy resistant, and the review of literature did not show large controlled investigations which would offer the most optimal treatment. Most frequently a rapid progression of the disease is seen.
