Farthmann B, Eberle J, Krasagakis K, Gstöttner M, Wang N, Bisson S, Orfanos C E
Department of Dermatology, University Medical Center Benjamin Franklin, The Free University of Berlin, Germany.
J Invest Dermatol. 1998 Mar;110(3):263-7. doi: 10.1046/j.1523-1747.1998.00131.x.
Reverse transcriptase polymerase chain reaction for the detection of tyrosinase-mRNA-positive cells in peripheral blood of melanoma patients, as a possible marker of hematogenous dissemination, has demonstrated varying detection rates. This study examined the sensitivity and reproducibility of the technique using a protocol of multiple polymerase chain reaction to determine circulating melanocytic cells. For each of the 123 melanoma patients included in this study, four nested polymerase chain reactions were performed from two blood specimens requiring both polymerase chain reactions from at least one blood sample to be positive to consider a patient as positive. Thus, a definitive result was obtained in 98% of the cases, whereas only 1.6% lacked conclusive findings. Thus, we found a correlation between the tyrosinase detection rate and the clinical stage. Circulating tyrosinase-mRNA-positive cells were detected in 13% of patients with primary tumor, 17% with regional skin/lymph node metastasis, and 44% with distant metastasis. Positivity also correlated with known melanoma progression markers such as gender, tumor thickness, and histologic type. Positive results were obtained more frequently in (i) men compared with women, (ii) patients with thick primary melanomas (> 4 mm: 38%) compared with those with thinner tumors (1.1-4 mm, 22%; < or = 1 mm, 5%), and (iii) patients with nonclassifiable (38%), nodular (34%), and occult primary melanomas (30%) compared with those with acrolentiginous (17%), superficial spreading (9%), or lentigo maligna melanoma (0%). These findings suggest that detection of tyrosinase-mRNA-positive cells in peripheral blood is not an adequate marker for identifying melanoma patients with distant metastasis. Reverse transcriptase polymerase chain positivity in early melanoma stages, however, as corresponding to other prognostic parameters, may indicate increased risk for the development of hematogenous metastasis and may be of value as a progression marker.
逆转录聚合酶链反应用于检测黑色素瘤患者外周血中酪氨酸酶 - mRNA阳性细胞，作为血行播散的一种可能标志物，其检测率各不相同。本研究使用多重聚合酶链反应方案来检测循环黑色素细胞，以检验该技术的敏感性和可重复性。对于本研究纳入的123例黑色素瘤患者中的每一例，从两份血液标本中进行四次巢式聚合酶链反应，要求至少一份血液样本的聚合酶链反应呈阳性才能将患者判定为阳性。因此，98%的病例获得了明确结果，而只有1.6%缺乏确凿的发现。我们发现酪氨酸酶检测率与临床分期之间存在相关性。在原发性肿瘤患者中，13%检测到循环酪氨酸酶 - mRNA阳性细胞；区域皮肤/淋巴结转移患者中为17%；远处转移患者中为44%。阳性结果也与已知的黑色素瘤进展标志物相关，如性别、肿瘤厚度和组织学类型。在以下情况中更频繁地获得阳性结果：（i）男性高于女性；（ii）原发性黑色素瘤厚（>4mm：38%）的患者高于肿瘤较薄（1.1 - 4mm，22%；≤1mm，5%）的患者；（iii）不可分类（38%）、结节状（34%）和隐匿性原发性黑色素瘤（30%）的患者高于肢端雀斑样痣型（17%）、浅表扩散型（9%）或恶性雀斑样痣黑色素瘤（0%）的患者。这些发现表明，外周血中酪氨酸酶 - mRNA阳性细胞的检测并非识别远处转移黑色素瘤患者的充分标志物。然而，早期黑色素瘤阶段的逆转录聚合酶链反应阳性，与其他预后参数相对应，可能表明血行转移发生风险增加，并且可能作为一种进展标志物具有价值。
