Schmidt H, Sørensen B S, von der Maase H, Bang C, Agger R, Hokland M, Nexo E
Department of Oncology, Aarhus University Hospital, Denmark.
Melanoma Res. 2002 Dec;12(6):585-92. doi: 10.1097/00008390-200212000-00008.
Circulating malignant cells in peripheral blood are thought to be precursors and surrogate markers of distant metastases and hence markers of a poor clinical outcome. In this study, we used the detection of MART-1 and tyrosinase (TYR) mRNA with a quantitative reverse transcription-polymerase chain reaction (RT-PCR) assay to identify circulating melanoma cells. Blood samples were obtained from 35 patients with metastatic melanoma before, during and after treatment with interleukin-2, interferon-alpha and cisplatin. In addition, MART-1 and TYR protein was identified by immunohistochemistry in consecutive biopsies from 15 of the patients. Analysis of three daily blood samples for 3 days demonstrated that four out of 11 patients examined were negative for both markers on all occasions, and two patients were positive for both markers on all occasions but one. The remaining five patients showed sporadic low positive results for one or the other of the two markers. By comparing the immunohistochemistry results from consecutive biopsies with the RT-PCR results, we demonstrated that patients with MART-1 and TYR protein in their tumour cells had circulating MART-1 and TYR mRNA in 77% and 54% of the cases, respectively. During treatment, the majority of patients who were positive for MART-1 and TYR mRNA converted to being negative. However, these conversions did not significantly correlate with objective response. The presence of TYR mRNA in one of the first two samples showed a trend towards being an independent prognostic factor for poor survival.
外周血中的循环恶性细胞被认为是远处转移的前体和替代标志物，因此也是临床预后不良的标志物。在本研究中，我们使用定量逆转录-聚合酶链反应（RT-PCR）检测MART-1和酪氨酸酶（TYR）mRNA，以鉴定循环中的黑色素瘤细胞。从35例转移性黑色素瘤患者在接受白细胞介素-2、干扰素-α和顺铂治疗前、治疗期间及治疗后采集血样。此外，通过免疫组织化学在15例患者的连续活检组织中鉴定MART-1和TYR蛋白。对11例患者连续3天每天采集的血样进行分析，结果显示，在所有检测时间点，11例接受检测的患者中有4例两种标志物均为阴性，2例患者除一次检测外，其余所有检测时间点两种标志物均为阳性。其余5例患者两种标志物中的一种或另一种呈现散发性低阳性结果。通过将连续活检组织的免疫组织化学结果与RT-PCR结果进行比较，我们发现肿瘤细胞中存在MART-1和TYR蛋白的患者，其循环中的MART-1和TYR mRNA阳性率分别为77%和54%。在治疗期间，大多数MART-1和TYR mRNA阳性的患者转变为阴性。然而，这些转变与客观反应并无显著相关性。在前两个样本中的一个样本中检测到TYR mRNA，显示出作为生存不良独立预后因素的趋势。
