The detection of isolated tumor cells in bone marrow comparing bright-field immunocytochemistry and multicolor immunofluorescence
The detection of isolated tumor cells in bone marrow comparing bright-field immunocytochemistry and multicolor immunofluorescence
复制标题
DOI：
10.1245/aso.2005.12.004
复制
发表时间：
2005-09-01
影响因子：
3.7
通讯作者：
Krag, M
中科院分区：
文献类型：
--
作者：
Krag, DN;Kusminsky, R;Krag, M
背景资料：据报道，通过免疫细胞化学（ICC）检测骨髓中分离的肿瘤细胞可预测早期乳腺癌的进展。最常见的染色程序使用亮视野ICC与细胞角蛋白（CK）抗体标记分离的肿瘤细胞。然而，这种方法可能导致假阳性染色事件。我们使用免疫荧光（IF）开发一个更具体的检测孤立的肿瘤细胞在骨髓样本中检测乳腺癌patients.Methods：我们比较ICC和IF并排检测乳腺癌患者的骨髓穿刺液中的癌细胞和假阳性染色事件，骨髓从健康供体，健康供体血液加标癌细胞。对于两种方法，分离的肿瘤细胞检测的主要靶标是CK。结果：乳腺癌患者骨髓穿刺液中CK+事件的检出率ICC组为18/31（58%），IF组为21/31（68%）。然而，在IF中，21例CK+病例中有17例被HC标记物染色，因此被鉴定为假阳性事件。在健康供体血液和骨髓中观察到令人惊讶的高CK+事件率。在所有健康供体样本中，CK+事件很容易通过IF鉴定为HC。两种方法在供体血液中的加标癌细胞的检测灵敏度相似。结论：血液和骨髓中CK+事件的频率很高，重要的是要注意，这是观察到的患者和那些没有癌症。具有多个HC标志物的IF允许直接区分造血和非造血来源的CK+细胞。
Background: The detection of isolated tumor cells in bone marrow by immunocytochemistry (ICC) has been reported to predict progression of early-stage breast cancer. The most common staining procedure uses bright-field ICC with cytokeratin (CK) antibodies to label isolated tumor cells. However, this method can result in false-positive staining events. We used multicolor immunofluorescence (IF) to develop a more specific assay for detecting isolated tumor cells in marrow samples from breast cancer patients.Methods: We compared ICC and IF side by side for detection of cancer cells and false-positive staining events on bone marrow aspirates from breast cancer patients, bone marrow from healthy donors, and healthy donor blood spiked with cancer cells. The primary target for isolated tumor cell detection was CK for both methods. IF used an additional set of antibodies to label hematopoietic cells (HCs).Results: The detection rate of CK+ events in breast cancer patient bone marrow aspirates was 18 (58%) of 31 for ICC and 21 (68%) of 31 for IF. However, with IF, 17 of 21 CK+ cases were stained with HC markers and thus were identified as false-positive events. A surprisingly high CK+ event rate was observed in healthy donor blood and marrow. In all healthy donor samples, CK+ events were readily identified as HCs by IF. Detection sensitivity of spiked cancer cells in donor blood was similar for both methods.Conclusions: There is a high frequency of CK+ events in blood and marrow, and it is important to note that this is observed both in patients with and those without cancer. IF with multiple HC markers allows straightforward discrimination between CK+ cells of hematopoietic and nonhematopoietic origin.