Detection of melanoma cells in peripheral blood and sentinel lymph nodes by RT-PCR analysis: A comparative study with immunohistochemistry
Detection of melanoma cells in peripheral blood and sentinel lymph nodes by RT-PCR analysis: A comparative study with immunohistochemistry
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DOI：
10.1177/030089160008600422
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发表时间：
2000-07-01
期刊：
影响因子：
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通讯作者：
Bianchi, R
中科院分区：
文献类型：
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作者：
Pellegrino, D;Bellina, CR;Bianchi, R
淋巴结转移的存在是预测黑色素瘤患者复发或生存的最佳促诊断因素。已经证明黑色素瘤转移通过引流肿瘤的第一个淋巴结扩散淋巴结转移（sentinel lymph node，SN）与淋巴系统的关系，外周血中黑色素瘤细胞的检测与黑色素瘤的预后直接相关。我们开发了一种单步逆转录-聚合酶链反应（RT-PCR）检测两种黑色素瘤特异性标志物：编码与黑色素合成相关的酶的酪氨酸酶基因，以及存在于肿瘤浸润性T淋巴细胞中的黑色素瘤抗原相关的T细胞。该方法在10（7）个淋巴细胞的背景中检测到两个肿瘤细胞。30名I-IV期皮肤黑色素瘤患者进入研究。在转移性患者中，在手术前、切除原发性黑色素瘤病变和SN或全淋巴结切除术后一个月、化疗开始前和此后每三个月采集血液样本。收集22例患者的SN，通过RT-PCR和常规病理学及免疫组化检测进行分析。初步结果表明，RT-PCR检测黑色素瘤标志物是一种敏感和有价值的方法，用于检测微转移和早期诊断和分期的黑色素瘤。
The presence of lymph node metastases is the best proanostic factor for predicting relapse or survival in melanoma patients. It has been demonstrated that melanoma metastases spread through the first lymph node(s) draining the tumor (sentinel lymph node, SN) to the lymphatic system and that detection of melanoma cells in peripheral blood directly correlates with prognosis in melanoma.To identify lymph node metastases and circulating melanocytes, we developed a single-step reverse transcriptase-polymerase chain reaction assay (RT-PCR) for detection of two melanoma-specific markers: the tyrosinase gene, which encodes an enzyme associated with melanin synthesis, and melanoma antigen-related T-cells, which are present in tumor infiltrating T-lymphocytes. This method detects two tumor cells in a background of 10(7) lymphocytes.Thirty patients with stage I-IV cutaneous melanoma entered the study. Blood samples were taken preoperatively, one month after excision of the primary melanoma lesion and the SN or total lymphadenectomy, and before the start of chemotherapy and every three months thereafter in metastatic patients. SNs were collected from 22 patients, bisected and analyzed by RT-PCR and routine pathological and immunohistochemical tests. The preliminary results indicate that RT-PCR for melanoma markers is a sensitive and valuable method for the detection of micrometastases and for early diagnosis and staging of melanoma.