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Clinical & Experimental Metastasis

, Volume 33, Issue 6, pp 551–562 | Cite as

Endoscopy-guided orthotopic implantation of colorectal cancer cells results in metastatic colorectal cancer in mice

  • Dominik Bettenworth
  • Marcus M. Mücke
  • Katrin Schwegmann
  • Andreas Faust
  • Christopher Poremba
  • Michael Schäfers
  • Dirk Domagk
  • Philipp LenzEmail author
Research Paper

Abstract

Advanced stage colorectal cancer (CRC) is still associated with limited prognosis. For preclinical evaluation of novel therapeutic approaches, murine models with orthotopic tumor growth and distant metastases are required. However, these models usually require surgical procedures possibly influencing tumor immunogenicity and development. The aim of this study was to establish a minimal-invasive endoscopy-based murine orthotopic model of metastatic CRC. During colonoscopy of CD-1 nude and non-obese diabetic/severe combined immunodeficiency (NOD/SCID) mice, implantation of Caco-2 and HT-29 CRC cells was performed subcutaneously (s.c.) or orthotopic into the colonic submucosa. White light endoscopy (WLE) and fluorescence endoscopy (FE) were applied for tumor detection in vivo. Ex vivo, resected tumors were examined by fluorescence reflectance imaging (FRI), histology, gelatin zymography and immunohistochemistry. In CD-1 nude mice, marked tumor growth was observed within 14 days after subcutaneous implantation while submucosal implantation failed to induce CRC after 17 weeks. In contrast, in NOD/SCID mice submucosal injection of HT-29 cells resulted in pronounced tumor growth 12 days post injectionem. Subsequently, rapid tumor expansion occurred, occupying the entire colonic circumference. Importantly, post mortem histological analyses confirmed liver metastases in 28.6 % and peritoneal metastases in 14.3 % of all mice. FRI and gelatin zymography did not detect a significantly increased matrix metalloproteinases (MMPs) expression in s.c. implanted tumors while MMP-tracer uptake was significantly enhanced in orthotopic implanted tumors. Neither s.c. nor orthotopic Caco-2 cell implantation resulted in tumor development. We successfully established an endoscopy-based model of metastatic CRC in immunodeficient mice.

Keywords

Colorectal cancer Metastasis Matrix metalloproteinase Fluorescence reflectance imaging Fluorescence endoscopy Endoscopy 

Abbreviations

CRC

Colorectal cancer

MMP

Matrix metalloproteinase

FRI

Fluorescence reflectance imaging

FE

Fluorescence endoscopy

NOD

Non-obese diabetic

SCID

Severe combined immunodeficiency

S.C.

Subcutaneous

NIR

Near infrared

MMPI

Matrix metalloproteinases inhibitor

G

Gauge

HE

Hematoxylin eosin

IL

Interleukin

TNF

Tumor necrosis factor

ROI

Region of interest

Notes

Acknowledgments

We thank S. Dufentester, E. Weber, I. Hoppe, C. Möllmann and D. Reinhardt for their expert technical assistance. This work was supported by an interdisciplinary grant from the Else-Kröner-Fresenius-Stiftung (2012_A94 to PL, DB, DD and MS). P. Lenz was supported by a research fellowship from the Deutsche Forschungsgemeinschaft, collaborative research center SFB 656, Münster, Germany. We thank Faekah Gohar for proofreading the manuscript. D. Bettenworth and P. Lenz were supported by a research fellowship from the Faculty of Medicine, Westfälische Wilhelms-Universität Münster.

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Copyright information

© Springer Science+Business Media Dordrecht 2016

Authors and Affiliations

  • Dominik Bettenworth
    • 1
  • Marcus M. Mücke
    • 1
  • Katrin Schwegmann
    • 2
  • Andreas Faust
    • 2
  • Christopher Poremba
    • 3
  • Michael Schäfers
    • 2
    • 4
  • Dirk Domagk
    • 1
    • 5
  • Philipp Lenz
    • 2
    • 6
    Email author
  1. 1.Department of Medicine BUniversity Hospital of MünsterMünsterGermany
  2. 2.European Institute for Molecular ImagingUniversity of MünsterMünsterGermany
  3. 3.Institute of Pathology Munich-NorthMunichGermany
  4. 4.Cells-in-Motion Cluster of Excellence (EXC 1003 – CiM)University of MünsterMünsterGermany
  5. 5.Josephs-Hospital WarendorfWarendorfGermany
  6. 6.Institute of Palliative CareUniversity Hospital of MünsterMünsterGermany

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