Detection of melanoma cells in the blood of melanoma patients by melanoma-inhibitory activity (MIA) reverse transcription-PCR.

摘要: The detection of tumor-specific mRNA transcripts in the blood patients by reverse transcription (RT)-PCR has been used as a very sensitive technique for determining systemically disseminated tumor cells. On basis previous expression studies, we aimed to trace melanoma cells RT-PCR melanoma-inhibitory activity (MIA) mRNA. To detect sensitively MIA total RNA isolated from peripheral mononuclear (PBMCs), established PCR-ELISA system. With this assay, detected one cell 2 ml single round 32 PCR cycles. A 295 PBMC samples 166 with melanocytic tumors were tested RT-PCR-ELISA: (a) 58 (99 samples) malignant melanomas stage I; (b) 49 (65 II; and (c) 47 (116 metastasized (stages III IV), an additional 12 (15 benign nevi. Forty-four (26.8%) 164 stages I II positive mRNA; III/IV, 33 (28.4%) 116 patients, irrespective clinically evident disease, positive. Eleven (84.6%) 13 disease without treatment mRNA-positive contrast only 19 (25.7%) 74 IV metastasis during chemotherapy. Furthermore, none 16 detectable metastases at that time point chemotherapy was mRNA-positive. Interestingly, 44 II, 20 still when retested after complete excision tumor. Our results reveal amplification PBMCs provides useful means systemic circulation. correlation between burden detected, and, addition, significant effect respect reduction number spread observed. However, seems be little value surrogate marker clinical staging or metastatic disease.