Clinical & Experimental Metastasis

, Volume 27, Issue 4, pp 261–271 | Cite as

A sensitive polymerase chain reaction-based method for detection and quantification of metastasis in human xenograft mouse models

  • Anastasia MalekEmail author
  • Carlo V. Catapano
  • Frank Czubayko
  • Achim Aigner
Research Paper


Tumor cell dissemination to distant organs accounts for the majority of cancer related deaths. Analysis of the stepwise process of metastasis formation and progression might provide novel therapeutic strategies for the treatment of disseminated cancer. However, studies with both biological and therapeutic endpoints would require highly sensitive and specific methods for precise quantification of the metastatic tumor burden in vivo. We have developed a quantitative real-time PCR-based assay for the detection and quantification of human tumor cells disseminated in mouse organs. The method relies on the parallel amplification of unique, species-specific, conserved and non-transcribed sequences in the mouse and human genomes. We tested the method in xenograft models to assess the metastatic potential of various cancer cell lines, the impact of injection modality and cell type on organ distribution, and the early stages of metastasis implantation and progression. With this method, we observed clear quantitative differences among colon cancer cell lines in terms of metastasis formation in the lung, consistent with the different in vitro growth properties. The mode of cell implantation and cell intrinsic properties strongly affected the metastatic pattern of prostate and breast cancer cell lines in mouse organs. The qPCR assay accurately determined the malignant cell burden even at early stages of metastasis progression in the lung. We describe a very sensitive assay for the highly reproducible detection and accurate quantification of human metastatic cells in mouse tissues and demonstrate its broad applicability to various experimental settings.


Metastasis Tumor xenografts Real-time PCR Colon cancer Prostate cancer Breast cancer Melanoma 



This work was supported by a grant from the Deutsche Forschungsgemeinschaft (DFG Forschergruppe 627 ‘Nanohale’) to F.C. and A.A. (AI 24/6-1), and grants from the Canton Ticino DECS and the Fondazione Ticinese Ricerca sul Cancro to C.V.C.


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Copyright information

© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  • Anastasia Malek
    • 1
    Email author
  • Carlo V. Catapano
    • 1
  • Frank Czubayko
    • 2
  • Achim Aigner
    • 2
  1. 1.Laboratory of Experimental OncologyOncology Institute of Southern SwitzerlandBellinzonaSwitzerland
  2. 2.Department of Pharmacology and ToxicologyPhilipps-University, School of MedicineMarburgGermany

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