Analyzing the Role of Proteases in Breast Cancer Progression and Metastasis Using Primary Cells from Transgenic Oncomice

Methods Mol Biol. 2021:2294:275-293. doi: 10.1007/978-1-0716-1350-4_20.


It is becoming increasingly evident that progression and metastasis of solid cancers is driven by the interaction of oncogene-transformed cancer cells and non-malignant host cells in the tumor stroma. In this process, the immune system contributes a complex set of highly important pro- and antitumor effects, which are not readily recapitulated by commonly used xenograft cancer models in immunodeficient mice.Therefore, we provide protocols for isolation of primary tumor cells from the MMTV-PymT mouse model for metastasizing breast cancer and their resubmission to congenic immunocompetent mice by orthotopic transplantation into the mammary gland or different routes of injection to induce organ-specific experimental metastasis, including intravenous, intracardiac, and caudal artery injection of tumor cells. Moreover, we describe protocols for sensitive detection and quantification of the metastatic burden.

Keywords: Bone metastasis; Brain metastasis; Breast cancer; Cathepsin; Experimental metastasis; Lung metastasis; Mammary adenocarcinoma; Mouse model; Orthotopic transplantation; Polyoma virus middle T antigen; Protease.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenocarcinoma / enzymology
  • Adenocarcinoma / pathology*
  • Animals
  • Brain Neoplasms / secondary*
  • Female
  • Lung Neoplasms / secondary*
  • Mammary Neoplasms, Experimental / enzymology
  • Mammary Neoplasms, Experimental / pathology*
  • Mice
  • Peptide Hydrolases / metabolism*
  • Transgenes
  • Tumor Cells, Cultured
  • Xenograft Model Antitumor Assays / methods*


  • Peptide Hydrolases