Department of Obstetrics and Gynaecology, University of Tübingen, Calwerstraße 7, 72076 Tubingen, Germany.
Breast Cancer Res Treat. 2011 Dec;130(3):833-44. doi: 10.1007/s10549-011-1710-0. Epub 2011 Aug 21.
The potential advantage of using quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) methodology to detect metastasis in sentinel lymph nodes (SLNs) of breast cancer (BC) patients was evaluated in this prospective study. We measured the expression of relevant gene transcripts in SLNs using an innovative algorithm and compared the results of single-marker assays versus multi-marker assays with conventional histological detection methods. SLNs from women aged ≥ 18 years diagnosed with unilateral BC were examined by haematoxylin-eosin staining and immunohistochemistry and analysed for transcripts of several relevant genes using qRT-PCR (learning group). Four candidate panels of expressed transcript combinations with high sensitivity and specificity were selected for further investigation. The candidate panels were then validated using SLNs from a second group of BC patients (validation group). In the learning group, 74/314 SLN sections from 150 patients were positive for metastasis by histology. The transcripts analysed showed the following individual sensitivities/specificities: cytokeratin 19 (CK19) 94.6%/97.9%; mammaglobin 1 (MGB1) 82.4%/91.7%; mammaglobin 2 (MGB2) 82.4%/96.7%; carcinoembryonic antigen (CEA) 71.6%/97.5%; EPCAM (epithelial cell adhesion molecule) 91.9%/97.1%; and NY-BR-1 82.4%/93.8%. The optimal panel based on the predefined criteria comprised four markers: CK19, MGB1, EPCAM, and NY-BR-1, of which ≥ 2 had to be positive (95.9% sensitivity, 95.0% specificity, 85.5% positive predictive value (PPV), and 98.7% negative predictive value (NPV)). Overall concordance with histology was 95.2%. In the validation group, 84/315 SLN sections from 235 patients were histologically positive, and panel sensitivity, specificity and overall accuracy were 88.1, 95.2 and 93.3%, respectively, at the SLN section level. In conclusion, molecular staging using expression patterns of relevant transcripts in SLNs could serve as a useful complement to standard diagnostic work-up in BC patients. The proposed flexible multi-parametric approach does not improve the overall accuracy compared with the single-marker approach. However, it overcomes several limitations of the previously reported molecular assays for SLN diagnosis.
本前瞻性研究旨在评估使用定量逆转录聚合酶链反应 (qRT-PCR) 方法检测乳腺癌 (BC) 患者前哨淋巴结 (SLN) 转移的潜在优势。我们使用创新算法测量了 SLN 中相关基因转录本的表达，并比较了单标记检测与传统组织学检测方法的多标记检测的结果。通过苏木精-伊红染色和免疫组织化学检查了年龄≥18 岁被诊断为单侧 BC 的女性的 SLN，并使用 qRT-PCR 分析了几种相关基因的转录本（学习组）。选择了具有高灵敏度和特异性的四个候选表达转录本组合面板进行进一步研究。然后使用来自第二组 BC 患者的 SLN（验证组）验证候选面板。在学习组中，通过组织学检查，在 150 名患者的 314 个 SLN 切片中有 74 个为阳性。分析的转录本显示出以下个体敏感性/特异性：细胞角蛋白 19 (CK19) 94.6%/97.9%；乳球蛋白 1 (MGB1) 82.4%/91.7%；乳球蛋白 2 (MGB2) 82.4%/96.7%；癌胚抗原 (CEA) 71.6%/97.5%；上皮细胞黏附分子 (EPCAM) 91.9%/97.1%；和 NY-BR-1 82.4%/93.8%。基于预设标准的最佳面板由四个标志物组成：CK19、MGB1、EPCAM 和 NY-BR-1，其中≥2 个标志物必须为阳性（95.9%的敏感性、95.0%的特异性、85.5%的阳性预测值 (PPV) 和 98.7%的阴性预测值 (NPV)）。总体与组织学的一致性为 95.2%。在验证组中，235 名患者中有 84 个 SLN 切片组织学阳性，在 SLN 切片水平上，面板敏感性、特异性和总准确性分别为 88.1%、95.2%和 93.3%。总之，使用 SLN 中相关转录本的表达模式进行分子分期可能成为 BC 患者标准诊断的有用补充。与单标记方法相比，所提出的灵活的多参数方法并没有提高整体准确性。然而，它克服了以前报道的用于 SLN 诊断的分子检测方法的几个局限性。
