Krag David N, Kusminsky Roberto, Manna Edward, Weaver Donald, Harlow Seth P, Covelli Michael, Stanley Mary A, McCahill Laurence, Ittleman Frank, Leavitt Bruce, Krag Martin, Amarante Patricia
Department of Surgery, University of Vermont College of Medicine, Burlington, VT 05401, USA.
Appl Immunohistochem Mol Morphol. 2009 Oct;17(5):403-8. doi: 10.1097/PAI.0b013e31819f4d6b.
Detection of disseminated tumor cells in the bone marrow may provide important prognostic information in breast cancer patients. With few exceptions the number of stained cells scored as cancer is very low; there may be only 1 cell per slide. This makes definitive interpretation of cancer in marrow challenging. False-positive staining of marrow cells with cytokeratin (CK) antibody is relatively common and makes interpretation more difficult. In this report we focus on false-positive staining of marrow specimens from breast cancer patients and noncancer controls and demonstrate that the frequency of false-positive events is common. Bone marrow was collected from 23 cancer-free donors and 60 breast cancer patients. Samples were processed by Ficoll density gradient centrifugation and slides were prepared for immunocytochemical staining with CK and irrelevant (IR) antibody. Slides were evaluated manually and positive cells were categorized as tumor cells, hematopoetic cells, or questionable cells. False-positive staining events were commonly observed in noncancer cases stained with CK or IR antibodies and in breast cancer cases stained with IR antibody. There was little difference in the number of breast cancer marrow specimens scored as tumor cells regardless of whether the antibody used was CK or IR. It is important to devise improved criteria and methods for accurate detection and interpretation of disseminated tumor cells in the marrow of breast cancer patients.
骨髓中播散肿瘤细胞的检测可为乳腺癌患者提供重要的预后信息。除少数例外情况外,被判定为癌细胞的染色细胞数量非常少;每张玻片上可能只有1个细胞。这使得对骨髓中癌细胞的明确判读具有挑战性。骨髓细胞被细胞角蛋白(CK)抗体误染色相对常见,这使得判读更加困难。在本报告中,我们聚焦于乳腺癌患者和非癌对照者骨髓标本的误染色情况,并证明误阳性事件的发生率很常见。从23名无癌供者和60名乳腺癌患者采集骨髓。样本经Ficoll密度梯度离心处理,玻片制备用于用CK和无关(IR)抗体进行免疫细胞化学染色。玻片经人工评估,阳性细胞被分类为肿瘤细胞、造血细胞或可疑细胞。在使用CK或IR抗体染色的非癌病例以及使用IR抗体染色的乳腺癌病例中,均常见误阳性染色事件。无论使用的抗体是CK还是IR,判定为肿瘤细胞的乳腺癌骨髓标本数量几乎没有差异。制定改进的标准和方法以准确检测和判读乳腺癌患者骨髓中的播散肿瘤细胞非常重要。