Visús Carmen, Andres Raquel, Mayordomo Jose I, Martinez-Lorenzo María J, Murillo Laura, Sáez-Gutiérrez Berta, Diestre Clara, Marcos Ivan, Astier Pilar, Godino Javier, Carapeto-Marquez de Prado Francisco J, Larrad Luis, Tres Alejandro
Division of Medical Oncology, Hospital Clínico Universitario Lozano Blesa, Zaragoza, Spain.
Melanoma Res. 2007 Apr;17(2):83-9. doi: 10.1097/CMR.0b013e3280a60878.
A need for factors predictive of prognosis is present in patients who are diagnosed with malignant melanoma. The detection of circulating melanoma cells by reverse transcriptase-polymerase chain reaction for tyrosinase mRNA is a possible negative prognostic factor. The aim of this study was to assess the prognostic value of reverse transcriptase-PCR for tyrosinase mRNA in peripheral blood samples. From January 2000 to February 2003, duplicate blood samples were drawn from 114 melanoma patients following surgery and informed consent, and were tested with reverse transcriptase-PCR, for tyrosinase mRNA. Outer primers for the first PCR were R1 (sense): TTGGCAGATTGTCTGTAGCC and R2 (antisense): AGGCATTGTGCATGCTGCT. For the second round of PCR, nested primers were R3 (sense): GTCTTTATGCAATGGAACGC and R4 (antisense): GCTATCCCAGTAAGTGGACT. Threshold for detection of the technique was determined by adding serially diluted MelJuSo cells to healthy volunteer blood samples. Overall, 91 (79.1%) patients tested negative for tyrosinase mRNA and 24 (20.9%) tested positive. The number of patients who tested positive by stage was 3/38 (7.9%) for stage I, 3/22 (13.6%) for stage II, 5/30 (16.7%) for stage III and 13/24 (54.2%) for stage IV (P< 0.0001). 11/90 (12.2%) patients with no evidence of disease (stage I, II and III) tested positive and 13/24 (54.2%) patients with clinically confirmed distant metastases (stage IV) tested positive (P<0.00001). With median follow-up of 372 days or to death (range: 0-1303 days), median progression-free survival has not been reached for tyrosinase-negative patients and was 265 days for tyrosinase-positive patients (P<0.00001, log-rank test=21.07). Median overall survival was 344 days for tyrosinase-positive patients and has not been reached for tyrosinase-negative patients (P=0.0001, log-rank test=21.38). Stage, Breslow thickness and result of RT-PCR were significant prognostic factors for disease-free survival in a multivariate analysis, and stage was the only significant prognostic factor for overall survival. In conclusion, detection of circulating melanoma cells by reverse transcriptase-PCR for tyrosinase mRNA is a significant adverse prognostic factor for disease-free survival in patients with malignant melanoma.
对于被诊断为恶性黑色素瘤的患者而言，存在对预后预测因素的需求。通过逆转录聚合酶链反应检测酪氨酸酶mRNA来检测循环黑色素瘤细胞是一个可能的不良预后因素。本研究的目的是评估逆转录聚合酶链反应检测外周血样本中酪氨酸酶mRNA的预后价值。从2000年1月至2003年2月，在获得知情同意后，从114例黑色素瘤患者术后采集两份血样，并用逆转录聚合酶链反应检测酪氨酸酶mRNA。第一轮PCR的外侧引物为R1（正义链）：TTGGCAGATTGTCTGTAGCC和R2（反义链）：AGGCATTGTGCATGCTGCT。第二轮PCR的巢式引物为R3（正义链）：GTCTTTATGCAATGGAACGC和R4（反义链）：GCTATCCCAGTAAGTGGACT。通过向健康志愿者血样中逐次添加稀释的MelJuSo细胞来确定该技术的检测阈值。总体而言，91例（79.1%）患者酪氨酸酶mRNA检测为阴性，24例（20.9%）检测为阳性。按分期检测为阳性的患者数量为：I期3/38例（7.9%），II期3/22例（13.6%），III期5/30例（16.7%），IV期13/24例（54.2%）（P<0.0001）。11/90例（12.2%）无疾病证据（I、II和III期）的患者检测为阳性，13/24例（54.2%）临床确诊有远处转移（IV期）的患者检测为阳性（P<0.00001）。中位随访37
