Enrichment with anti-cytokeratin alone or combined with anti-EpCAM antibodies significantly increases the sensitivity for circulating tumor cell detection in metastatic breast cancer patients.
Enrichment with anti-cytokeratin alone or combined with anti-EpCAM antibodies significantly increases the sensitivity for circulating tumor cell detection in metastatic breast cancer patients.
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DOI：
10.1186/bcr2131
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发表时间：
2008
期刊：
影响因子：
--
通讯作者：
Burke JF
中科院分区：
文献类型：
--
作者：
Deng G;Herrler M;Burgess D;Manna E;Krag D;Burke JF
循环肿瘤细胞（CTCs）在大多数癌症患者中都可以检测到，它们可以满足现有的医学需求，在治疗过程中监测癌症患者并帮助确定复发性疾病。CTC在癌症患者的血液中很少发现，为了灵敏地检测CTC，需要富集。尽管CTC的鉴定标准是细胞角蛋白阳性（CK+）、CD45阴性（CD45-）和4'6-二氨基-2-苯基吲哚（核染色）阳性（DAPI+），但大多数CTC富集技术都是基于抗epcam抗体的。然而，一些肿瘤细胞表达低或不表达EpCAM。在这里，我们提出了一种高灵敏度和可重复性的富集方法，该方法基于单独结合抗ck或抗ck和抗epcam抗体的组合。使用CellSearch™系统（Veridex， LLC, rariitan， NJ， USA）处理49例转移性乳腺癌患者的血液样本，同时使用我们的CTC检测方法。我们单独使用抗ck或与抗epcam抗体联合使用CTC富集。使用亮场和荧光标记抗ck、抗cd45和DAPI（核染色）图像进行CTC鉴定。Ariol®系统（Genetix USA Inc, San Jose， CA， USA）用于玻片上ctc的自动细胞图像捕获和分析。我们的方法能够富集CK+&EpCAM+、CK+&EpCAM-/low和CK-/low&EpCAM+三种类型的ctc。在盲法比较中，我们的抗ck抗体富集方法在49例乳腺癌患者中显示出明显高于CellSearch™系统的CTC阳性率（49% vs. 29%）和更大的动态CTC检测范围（1 ~ 571 vs. 1 ~ 270）。在CTC计数较高的患者（每7.5 ml血液中有20个CTC）中，我们的方法比CellSearch™方法检测到的CTC多15%至111%。根据既定的CTC标准，Ariol®系统的三张荧光和明场图像减少了CTC假阳性事件的数量。我们的数据表明，肿瘤特异性细胞内CK标记可以用于有效的CTC富集。单独使用抗ck或联合使用抗epcam抗体可显著提高检测灵敏度。Ariol®系统的三个荧光和明场优越图像减少了假阳性CTC事件。
Circulating tumor cells (CTCs) are detectable in most cancer patients and they can meet an existing medical need to monitor cancer patients during a course of treatment and to help determine recurrent disease. CTCs are rarely found in the blood of cancer patients and enrichment is necessary for sensitive CTC detection. Most CTC enrichment technologies are anti-EpCAM antibody based even though CTC identification criteria are cytokeratin positive (CK+), CD45 negative (CD45-) and 4'6-diamidino-2-phenylindole (nuclear stain) positive (DAPI+). However, some tumor cells express low or no EpCAM. Here we present a highly sensitive and reproducible enrichment method that is based on binding to anti-CK alone or a combination of anti-CK and anti-EpCAM antibodies. Blood samples from 49 patients with metastatic breast cancer were processed using the CellSearch™ system (Veridex, LLC, Raritan, NJ, USA), in parallel with our CTC assay method. We used anti-CK alone or in combination with anti-EpCAM antibodies for CTC enrichment. Brightfield and fluorescence labeled anti-CK, anti-CD45 and DAPI (nuclear stain) images were used for CTC identification. The Ariol® system (Genetix USA Inc, San Jose, CA, USA) was used for automated cell image capture and analysis of CTCs on glass slides. Our method has the capability to enrich three types of CTCs including CK+&EpCAM+, CK+&EpCAM-/low, and CK-/low&EpCAM+ cells. In the blind method comparison, our anti-CK antibody enrichment method showed a significantly higher CTC positive rate (49% vs. 29%) and a larger dynamic CTC detected range (1 to 571 vs. 1 to 270) than that of the CellSearch™ system in the total of 49 breast cancer patients. Our method detected 15 to 111% more CTCs than the CellSearch™ method in patients with higher CTC counts (>20 CTCs per 7.5 ml of blood). The three fluorescent and brightfield images from the Ariol® system reduced the number of false-positive CTC events according to the established CTC criteria. Our data indicate that the tumor-specific intracellular CK marker could be used for efficient CTC enrichment. Enrichment with anti-CK alone or combined with anti-EpCAM antibodies significantly enhances assay sensitivity. The three fluorescent and brightfield superior images with the Ariol® system reduced false-positive CTC events.
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影响因子：
45.3
作者：
Cristofanilli, M;Hayes, DF;Terstappen, LWMM
通讯作者：
Terstappen, LWMM
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158.5
作者：
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4.5
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3.7
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