EPISPOT assay: detection of viable DTCs/CTCs in solid tumor patients.
EPISPOT assay: detection of viable DTCs/CTCs in solid tumor patients.
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DOI：
10.1007/978-3-642-28160-0_6
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发表时间：
2012-01-01
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通讯作者：
Alix-Panabieres, Catherine
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作者：
Alix-Panabieres, Catherine
外周血中循环肿瘤细胞（CTC）和骨髓中播散性肿瘤细胞（DTC）的计数和表征可以提供重要的预后信息，并可能有助于监测治疗效果。由于目前的测定不能区分凋亡和存活的DTC/CTC，因此现在可以应用新的ELISPOT测定（命名为“EPISPOT”），其检测从单个上皮癌细胞分泌/释放/脱落的蛋白质。将细胞在包被有抗体的膜上短时间培养，所述抗体捕获分泌/释放/脱落的蛋白，随后通过用荧光染料标记的二抗检测所述蛋白。在乳腺癌中，我们测量了细胞角蛋白-19（CK 19）和粘蛋白-1（MUC 1）的释放，并证明许多患者具有活性DTC，即使在明显局限性肿瘤的患者中（M（0）期：54%）。初步临床数据显示，DTC释放CK 19的患者预后不良。我们还研究了M1乳腺癌患者外周血中的CTC或CK 19分泌细胞，结果显示CK 19-SC患者的临床结局更差。在前列腺癌中，我们使用前列腺特异性抗原（PSA）分泌作为标志物，发现相当一部分CTC分泌成纤维细胞生长因子-2（FGF 2），这是一种已知的干细胞生长因子。总之，EPISPOT检测为检测和表征癌症患者中的活DTC/CTC提供了新的机会，并且可以扩展到揭示CTC/DTC蛋白指纹的多参数分析。
The enumeration and characterization of circulating tumor cells (CTCs) in the peripheral blood and disseminated tumor cells (DTCs) in bone marrow may provide important prognostic information and might help to monitor efficacy of therapy. Since current assays cannot distinguish between apoptotic and viable DTCs/CTCs, it is now possible to apply a novel ELISPOT assay (designated 'EPISPOT') that detects proteins secreted/released/shed from single epithelial cancer cells. Cells are cultured for a short time on a membrane coated with antibodies that capture the secreted/released/shed proteins which are subsequently detected by secondary antibodies labeled with fluorochromes. In breast cancer, we measured the release of cytokeratin-19 (CK19) and mucin-1 (MUC1) and demonstrated that many patients harbored viable DTCs, even in patients with apparently localized tumors (stage M(0): 54%). Preliminary clinical data showed that patients with DTC-releasing CK19 have an unfavorable outcome. We also studied CTCs or CK19-secreting cells in the peripheral blood of M1 breast cancer patients and showed that patients with CK19-SC had a worse clinical outcome. In prostate cancer, we used prostate-specific antigen (PSA) secretion as marker and found that a significant fraction of CTCs secreted fibroblast growth factor-2 (FGF2), a known stem cell growth factor. In conclusion, the EPISPOT assay offers a new opportunity to detect and characterize viable DTCs/CTCs in cancer patients and it can be extended to a multi-parameter analysis revealing a CTC/DTC protein fingerprint.