Letter | Published:

Tumour-cell-induced endothelial cell necroptosis via death receptor 6 promotes metastasis

Nature volume 536, pages 215218 (11 August 2016) | Download Citation


Metastasis is the leading cause of cancer-related death in humans. It is a complex multistep process during which individual tumour cells spread primarily through the circulatory system to colonize distant organs1,2,3. Once in the circulation, tumour cells remain vulnerable, and their metastatic potential largely depends on a rapid and efficient way to escape from the blood stream by passing the endothelial barrier4,5,6,7,8,9. Evidence has been provided that tumour cell extravasation resembles leukocyte transendothelial migration7,8,9. However, it remains unclear how tumour cells interact with endothelial cells during extravasation and how these processes are regulated on a molecular level. Here we show that human and murine tumour cells induce programmed necrosis (necroptosis) of endothelial cells, which promotes tumour cell extravasation and metastasis. Treatment of mice with the receptor-interacting serine/threonine-protein kinase 1 (RIPK1)-inhibitor necrostatin-1 or endothelial-cell-specific deletion of RIPK3 reduced tumour-cell-induced endothelial necroptosis, tumour cell extravasation and metastasis. In contrast, pharmacological caspase inhibition or endothelial-cell-specific loss of caspase-8 promoted these processes. We furthermore show in vitro and in vivo that tumour-cell-induced endothelial necroptosis leading to extravasation and metastasis requires amyloid precursor protein expressed by tumour cells and its receptor, death receptor 6 (DR6), on endothelial cells as the primary mediators of these effects. Our data identify a new mechanism underlying tumour cell extravasation and metastasis, and suggest endothelial DR6-mediated necroptotic signalling pathways as targets for anti-metastatic therapies.

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We thank S. Fulda and our friends and colleagues for comments on the manuscript. We also thank S. Hümmer for secretarial help and C. Ringel, J. Hoffmann, I.-M. Gross, D. Magalei and M. Winkels for technical help. This work was supported by the German Cancer Aid and the Max Planck Society. K.H. was supported by the China Scholarship Council. U.C.M. was supported by a grant from the Deutsche Forschungsgemeinschaft (MU 1457/9-2). M.P. received funding from the European Research Council (grant agreement 323040), the Deutsche Forschungsgemeinschaft (SFB670, SFB829), Worldwide Cancer Research (grant 15-0228) and the Helmholtz Alliance Preclinical Comprehensive Cancer Center.

Author information


  1. Max Planck Institute for Heart and Lung Research, Department of Pharmacology, Ludwigstrasse 43, 61231 Bad Nauheim, Germany

    • Boris Strilic
    • , Lida Yang
    • , Julián Albarrán-Juárez
    •  & Stefan Offermanns
  2. University of Cologne, Institute for Genetics, Center for Molecular Medicine (CMMC), and Cologne Excellence Cluster on Cellular Stress Responses in Aging-Associated Diseases (CECAD), Joseph-Stelzmann-Strasse 26, 50931 Cologne, Germany

    • Laurens Wachsmuth
    •  & Manolis Pasparakis
  3. University of Heidelberg, Department of Bioinformatics and Functional Genomics, Institute for Pharmacy and Molecular Biotechnology, Im Neuenheimer Feld 364, 69120 Heidelberg, Germany

    • Kang Han
    •  & Ulrike C. Müller
  4. J. W. Goethe University Frankfurt, Medical Faculty, Theodor-Stern-Kai 7, 60590 Frankfurt, Germany

    • Stefan Offermanns


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B.S. performed most of the experiments and analysed the data. L.Y. generated mice with a conditional Ripk3 allele and contributed to in vitro and in vivo studies. J.A.J. contributed to in vitro experiments. L.W. and M.P. generated MLKL-deficient animals. K.H. and U.C.M. purified APPsα and performed APP-related experiments. B.S. and S.O. designed the study, discussed data and wrote the manuscript. All authors commented on the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Boris Strilic or Stefan Offermanns.

Reviewer Information Nature thanks C. Betsholtz, S. Tavazoie and the other anonymous reviewer(s) for their contribution to the peer review of this work.

Extended data

Supplementary information

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  1. 1.

    Supplementary Figures

    This file contains Supplementary Figures 1-2, original source images for data obtained by electrophoretic separation.

Excel files

  1. 1.

    Supplementary Table 1

    This table contains information regarding the identification of endothelial receptors involved in the regulation of tumour cell-induced endothelial cell death (expression data, siRNA target sequences).


  1. 1.

    Examples of endothelial cells undergoing necroptosis after contact with a tumour cell

    Time lapse imaging of HUVEC upon contact with MDA-MB-231 tumour cells (GFP, green). Hoechst33342 labels all cell nuclei (blue) and EthD-III (red) labels necroptotic cells. Imaging shows endothelial cells that undergo necroptotic cell death (arrow head) several hours after contact with a tumour cell.

  2. 2.

    Examples of endothelial cells undergoing necroptosis after contact with a tumour cell

    Time lapse imaging of HUVEC upon contact with MDA-MB-231 tumour cells (GFP, green). Hoechst33342 labels all cell nuclei (blue) and EthD-III (red) labels necroptotic cells. Imaging shows endothelial cells that undergo necroptotic cell death (arrow head) several hours after contact with a tumour cell.

  3. 3.

    Examples of endothelial cells undergoing necroptosis after contact with a tumour cell

    Time lapse imaging of HUVEC upon contact with MDA-MB-231 tumour cells (GFP, green). Hoechst33342 labels all cell nuclei (blue) and EthD-III (red) labels necroptotic cells. Imaging shows endothelial cells that undergo necroptotic cell death (arrow head) several hours after contact with a tumour cell.

  4. 4.

    Morphological criteria for the distinction between living, apoptotic and necrotic endothelial cells

    Time lapse imaging of HUVEC cultured in the presence of PBS (control) or in the presence of H2O2 (1 mM) to induce necrosis or TNF (100 ng/ml) to induce apoptosis. Cell nuclei were stained with Hoechst33342 (blue). Nuclei of necrotic cells stained positive for the membrane-impermeant nuclear dye EthD-III (red). Living cells appear with normal round to kidney-shaped nuclei and are negative for EthD-III. Necrotic cells appear with normal round to kidney-shaped nuclei or with minor degrees of nuclear shrinkage (no condensation and no fragmentation) and are positive for EthD-III. Apoptotic cells appear with strong condensed and frequently fragmented nuclei and are negative for EthD-III. No late apoptotic cells are shown in the videos.

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