Multimarker quantitative real-time PCR detection of circulating melanoma cells in peripheral blood: Relation to disease stage in melanoma patients
Multimarker quantitative real-time PCR detection of circulating melanoma cells in peripheral blood: Relation to disease stage in melanoma patients
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DOI：
10.1373/clinchem.2004.045096
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发表时间：
2005-06-01
影响因子：
9.3
通讯作者：
Hoon, DSB
中科院分区：
文献类型：
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作者：
Koyanagi, K;Kuo, C;Hoon, DSB
背景：检测循环中的黑色素瘤细胞对评估肿瘤进展可能很重要。本研究的目的是建立一种特异、可靠的多标记实时定量逆转录聚合酶链式反应(QRT)方法，用于检测患者血液中黑色素瘤细胞。在优化研究中，我们测试了17个黑色素瘤细胞系和49个志愿者外周血白细胞(PBL)样本。我们进行了RNA和黑色素瘤细胞稀释研究，以评估检测限和分析的不精确度。我们检测了94例黑色素瘤患者[美国癌症联合委员会(AJCC)I期，20例；II期，20例；III期，32例；IV期，22例]的血液标本中的mRNAs。结果：在黑色素瘤细胞系中检测到所有标志物，而在志愿者的PBL中未检测到这些标志物。在黑色素瘤细胞稀释研究中，QRT法可在107个外周血淋巴细胞中检出1个黑色素瘤细胞。在AJCC I、II、III和IV期黑色素瘤患者中，分别有15%、30%、75%和86%的患者检测到标志物。阳性标志物的数量与AJCC分期显著相关(Spearman相关系数=0.58P<；0.0001)。血液中分子标志物的检测可能有助于黑色素瘤患者转移的检测和治疗反应的监测。(C)2005年美国临床化学协会
Background: Detection of melanoma cells in circulation may be important in assessing tumor progression. The objective of this study was to develop a specific, reliable multimarker quantitative real-time reverse transcription-PCR (qRT) assay for detecting melanoma cells in patients' blood.Methods: We developed qRT assays for the mRNA of four melanoma-associated markers: MART-1, Ga1NAc-T, PAX-3, and MAGE-A3. In optimization studies, we tested 17 melanoma cell lines and 49 peripheral blood leukocyte (PBL) samples from volunteers. We performed RNA and melanoma cell dilution studies to assess the detection limits and imprecision of the assays. We measured the mRNAs in blood specimens from 94 melanoma patients [American joint Committee on Cancer (AJCC) stage I, n 20; II, n = 20; III, n = 32; IV, n = 22].Results: All markers were frequently detected in melanoma cell lines, whereas none of the markers was detected in PBLs from volunteers. The qRT assay could detect 1 melanoma cell in 107 PBLs in the melanoma cell-dilution studies. Markers were detected in 15%, 30%, 75%, and 86% of melanoma patients with AJCC stage I, II, III, and IV disease, respectively. The number of positive markers and AJCC stage were significantly correlated (Spearman correlation coefficient = 0.58; P < 0.0001).Conclusions: Multimarker qRT can detect circulating melanoma cells in blood. Measurement of the studied molecular markers in blood may be useful in detection of metastasis and monitoring treatment response of melanoma patients. (c) 2005 American Association for Clinical Chemistry