Comparison of three molecular assays for the detection and molecular characterization of circulating tumor cells in breast cancer.
Comparison of three molecular assays for the detection and molecular characterization of circulating tumor cells in breast cancer.
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DOI：
10.1186/bcr3395
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发表时间：
2013-03-07
期刊：
影响因子：
--
通讯作者：
Lianidou ES
中科院分区：
文献类型：
--
作者：
Strati A;Kasimir-Bauer S;Markou A;Parisi C;Lianidou ES
迫切需要对循环肿瘤细胞 (CTC) 检测和分子表征的不同分析方法进行比较研究，因为在临床实践中使用之前，测定标准化至关重要。我们比较了三种不同的 CTC 分子检测方法。为了避免由于分析前错误而导致的差异，我们在整个研究过程中使用了相同的 cDNA。使用抗 EpCAM 和抗 MUC1 涂层磁珠从 254 名早期乳腺癌和 51 名转移性乳腺癌患者以及 30 名健康个体的 2 × 5 ml 外周血中分离出 CTC。通过以下方法分析相同的 cDNA：a) 针对 CK-19 的单重 RT-qPCR 测定； b) 针对 CK-19、HER-2、MAGE-A3 和 PBGD 的多重 RT-qPCR； c) 针对 GA733-2、MUC-1、HER-2 和 β-肌动蛋白的市售分子检测（AdnaTest BreastCancer）。在早期乳腺癌中，CK-19 RT-qPCR、多重 RT-qPCR 和 AdnaTest 分别在 14.2%、22.8% 和 16.5% 的受试者中呈 CTC 阳性。 AdnaTest 与 CK-19 RT-qPCR 之间的一致性为 72.4%，而 AdnaTest 与多重 RT-qPCR 之间的一致性为 64.6%。在有明显转移的患者中，CK-19 RT-qPCR、多重 RT-qPCR 和 AdnaTest 分别在 41.2%、39.2% 和 54.9% 的患者中呈阳性。 AdnaTest 与 CK-19 RT-qPCR 之间的一致性为 70.6%，而 AdnaTest 与多重 RT-qPCR 之间的一致性为 68.6%。所有 CTC 检测在大约 70% 的病例中给出相似的结果。在转移性环境中发现了更好的一致性，这可能是由于该组中较高的肿瘤负荷所致。不一致可能归因于用于评估 CTC 阳性的不同基因转录本。我们的结果表明 CTC 异质性对于通过不同分析方法进行检测的重要性。
Comparison studies between different analytical methodologies for circulating tumor cells (CTC) detection and molecular characterization are urgently needed, since standardization of assays is essential before their use in clinical practice. We compared three different CTC molecular assays. To avoid discrepancies due to pre-analytical errors we used the same cDNAs throughout our study. CTC were isolated using anti-EpCAM and anti-MUC1 coated magnetic beads from 2 × 5 ml of peripheral blood of 254 early and 51 metastatic breast cancer patients and 30 healthy individuals. The same cDNAs were analyzed by: a) singleplex RT-qPCR assay for CK-19; b) multiplex RT-qPCR for CK-19, HER-2, MAGE- A3, and PBGD; and c) a commercially available molecular assay (AdnaTest BreastCancer) for GA733-2, MUC-1, HER-2 and beta-actin. In early breast cancer, CK-19 RT-qPCR, multiplex RT-qPCR and the AdnaTest, were positive for the presence of CTC in 14.2%, 22.8% and 16.5% subjects, respectively. The concordance between the AdnaTest and CK-19 RT-qPCR was 72.4% while between the AdnaTest and multiplex RT-qPCR was 64.6%. In patients with overt metastasis, CK-19 RT-qPCR, multiplex RT-qPCR and the AdnaTest were positive in 41.2%, 39.2% and 54.9% patients, respectively. The concordance between the AdnaTest and CK-19 RT-qPCR was 70.6% while between the AdnaTest and multiplex RT-qPCR was 68.6%. All CTC assays gave similar results in about 70% of cases. Better agreement was found in the metastatic setting, possibly explained by the higher tumor load in this group. Discordances could be attributed to the different gene transcripts used to evaluate CTC positivity. Our results indicate the importance of CTC heterogeneity for their detection by different analytical methodologies.
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DOI：
10.1186/bcr2349
发表时间：
2009
期刊：
Breast cancer research : BCR
影响因子：
--
作者：
Fehm T;Hoffmann O;Aktas B;Becker S;Solomayer EF;Wallwiener D;Kimmig R;Kasimir-Bauer S
通讯作者：
Kasimir-Bauer S
影响因子：
158.5
作者：
Cristofanilli, M;Budd, GT;Hayes, DF
通讯作者：
Hayes, DF
DOI：
10.1186/bcr3099
发表时间：
2012-01-20
期刊：
Breast cancer research : BCR
影响因子：
--
作者：
Kasimir-Bauer S;Hoffmann O;Wallwiener D;Kimmig R;Fehm T
通讯作者：
Fehm T
影响因子：
11.5
作者：
Sieuwerts, Anieta M.;Mostert, Bianca;Martens, John W. M.
通讯作者：
Martens, John W. M.
影响因子：
78.8
作者：
Pantel, Klaus;Alix-Panabieres, Catherine;Riethdorf, Sabine
通讯作者：
Riethdorf, Sabine