Immunomagnetic enrichment, genomic characterization, and prognostic impact of circulating melanoma cells
Immunomagnetic enrichment, genomic characterization, and prognostic impact of circulating melanoma cells
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DOI：
10.1158/1078-0432.ccr-0424-03
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发表时间：
2004-01-15
影响因子：
11.5
通讯作者：
Klein, CA
中科院分区：
文献类型：
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作者：
Ulmer, A;Schmidt-Kittler, O;Klein, CA
目的：外周血中黑色素瘤细胞的发现，到目前为止主要是通过基于聚合酶链反应（PCR）对酪氨酸酶mRNA的检测推断出来的，它与转移性黑色素瘤有关。循环细胞的恶性性质及其预后意义均无法确定。为了解决这个问题，我们对免疫磁珠分离的循环黑色素瘤细胞进行了染色体畸变分析，并对入组患者进行了临床随访研究。
实验设计：在一项前瞻性研究中，采集了164例黑色素瘤患者和50例无恶性疾病的捐献者的血液样本。使用一种针对黑色素瘤相关硫酸软骨素蛋白聚糖的鼠单克隆抗体，通过免疫磁珠细胞分选富集循环黑色素瘤细胞。为了证明阳性细胞的恶性来源并确定其染色体畸变，我们通过单细胞比较基因组杂交（SCOMP）分析了来自7例患者的15个单独分离的细胞的基因组。
结果：循环黑色素瘤细胞的绝对频率和相对频率与分期以及是否存在可检测到的肿瘤有关。检测到两个或更多细胞与转移性黑色素瘤患者生存率降低显著相关。通过SCOMP分析的所有细胞都显示出多种染色体变化，并具有黑色素瘤的典型畸变。
结论：免疫磁富集能够分离循环黑色素瘤细胞并对其进行基因组特征分析。对转移性患者生存率的预后影响显然反映了正在进行的肿瘤扩散的侵袭性。对富集和分离的细胞进行直接基因组分析将有助于阐明全身性黑色素瘤形成的分子遗传基础。
Purpose: The finding of melanoma cells in the peripheral blood, thus far mainly inferred from the PCR-based demonstration of tyrosinase mRNA, has been associated with metastatic melanoma. Neither the malignant nature nor the prognostic significance of circulating cells could be established. To address this question, we analyzed immunomagnetically isolated circulating melanoma cells for chromosomal aberrations and performed a clinical follow-up study of the enrolled patients.Experimental Design: In a prospective study, blood samples were taken from 164 melanoma patients and 50 donors without malignant disease. Circulating melanoma cells were enriched by immunomagnetic cell sorting using a murine monoclonal antibody against the melanoma-associated chondroitin sulfate proteoglycan. To prove the malignant origin of the positive cells and to define their chromosomal aberrations, we analyzed the genomes of 15 individually isolated cells from seven patients by single-cell comparative genomic hybridization (SCOMP).Results: Absolute and relative frequencies of circulating melanoma cells were associated with stage and with the presence or absence of detectable tumor. The detection of two or more cells correlated significantly with a reduced survival of patients with metastatic melanoma. All of the cells that were analyzed by SCOMP displayed multiple chromosomal changes and carried aberrations typical for melanoma.Conclusions: Immunomagnetic enrichment enables isolation and genomic characterization of circulating melanoma cells. The prognostic impact on survival of metastatic patients apparently reflects the aggressiveness of an ongoing tumor spread. Direct genomic analysis of the enriched and isolated cells will help to clarify the molecular-genetic basis of the establishment of generalized melanoma.