Circulating tumour cells escape from EpCAM-based detection due to epithelial-to-mesenchymal transition

BMC Cancer

Tobias M Gorges¹  Ingeborg Tinhofer²  Michael Drosch¹  Lars Röse¹  Thomas M Zollner¹  Thomas Krahn¹  Oliver von Ahsen³ 

[1] Bayer Pharma AG, Muellerstr. 178, Berlin 13353, Germany

[2] Charite CCM, Klinik für Radioonkologie und Strahlentherapie Chariteplatz 1, Berlin 10117, Germany

[3] Boehringer Ingelheim Pharma GmbH & Co. KG Drug Metabolism&Pharmacokinetics, Biberach, Germany

关键词: Epithelial-mesenchymal transition;    Metastasis;    Xenograft;    Breast cancer;    Circulating tumour cells;   

Others  :  1080429
DOI  :  10.1186/1471-2407-12-178

 received in 2012-01-23, accepted in 2012-04-11,  发布年份 2012

【 摘 要 】

Background

Circulating tumour cells (CTCs) have shown prognostic relevance in metastatic breast, prostate, colon and pancreatic cancer. For further development of CTCs as a biomarker, we compared the performance of different protocols for CTC detection in murine breast cancer xenograft models (MDA-MB-231, MDA-MB-468 and KPL-4). Blood samples were taken from tumour bearing animals (20 to 200 mm²) and analysed for CTCs using 1. an epithelial marker based enrichment method (AdnaTest), 2. an antibody independent technique, targeting human gene transcripts (qualitative PCR), and 3. an antibody-independent approach, targeting human DNA-sequences (quantitative PCR). Further, gene expression changes associated with epithelial-to-mesenchymal transition (EMT) were determined with an EMT-specific PCR assay.

Methods

We used the commercially available Adna Test, RT-PCR on human housekeeping genes and a PCR on AluJ sequences to detect CTCs in xenografts models. Phenotypic changes in CTCs were tested with the commercially available “Human Epithelial to Mesenchymal Transition RT-Profiler PCR Array”.

Results

Although the AdnaTest detects as few as 1 tumour cell in 1 ml of mouse blood spiking experiments, no CTCs were detectable with this approach in vivo despite visible metastasis formation. The presence of CTCs could, however, be demonstrated by PCR targeting human transcripts or DNA-sequences - without epithelial pre-enrichment. The failure of CTC detection by the AdnaTest resulted from downregulation of EpCAM, whereas mesenchymal markers like Twist and EGFR were upregulated on CTCs. Such a change in the expression profile during metastatic spread of tumour cells has already been reported and was linked to a biological program termed epithelial-mesenchymal transition (EMT).

Conclusions

The use of EpCAM-based enrichment techniques leads to the failure to detect CTC populations that have undergone EMT. Our findings may explain clinical results where low CTC numbers have been reported even in patients with late metastatic cancers. These results are a starting point for the identification of new markers for detection or capture of CTCs, including the mesenchymal-like subpopulations.

【 授权许可】

   
2012 Gorges et al.; licensee BioMed Central Ltd.

【 图 表 】

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