Three-dimensional cultures modeling premalignant progression of human breast epithelial cells: Role of cysteine cathepsins

Stefanie R. Mullins, Mansoureh Sameni, Galia Blum, Matthew Bogyo, Bonnie F. Sloane, Kamiar Moin*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

27 Scopus citations

Abstract

The expression of the cysteine protease cathepsin B is increased in early stages of human breast cancer. To assess the potential role of cathepsin B in premalignant progression of breast epithelial cells, we employed a 3D reconstituted basement membrane overlay culture model of MCF10A human breast epithelial cells and isogenic variants that replicate the in vivo phenotypes of hyper plasia (MCF10AneoT) and atypical hyperplasia (MCF10AT1). MCF10A cells developed into polarized acinar structures with central lumens. In contrast, MCF10AneoT and MCF10AT1 cells form larger structures in which the lumens are filled with cells. CA074Me, a cell-permeable inhibitor selective for the cysteine cathepsins B and L, reduced proliferation and increased apoptosis of MCF10A, MCF10AneoT and MCF10AT1 cells in 3D culture. We detected active cysteine cathepsins in the isogenic MCF10 variants in 3D culture with GB111, a cell-permeable activitybased probe, and established differential inhibition of cathepsin B in our 3D cultures. We conclude that cathepsin B promotes proliferation and premalignant progression of breast epithelial cells. These findings are consistent with studies by others showing that deletion of cathepsin B in the transgenic MMTV-PyMT mice, a murine model that is predisposed to development of mammary cancer, reduces malignant progression.

Original languageAmerican English
Pages (from-to)1405-1416
Number of pages12
JournalBiological Chemistry
Volume393
Issue number12
DOIs
StatePublished - Dec 2012
Externally publishedYes

Bibliographical note

Funding Information:
Acknowledgements: This work was supported by U.S. Public Health Service Grant CA 56586 and a Department of Defense Breast Cancer Center of Excellence (DAMD17-02–1-0693). The Microscopy, Imaging and Cytometry Resources Laboratory was supported by National Institutes of Health Center Grants P30ES06639 and P30CA22453, a Roadmap Grant U54RR020843 and the Perinatal Research Branch of the National Institutes of Health.

Keywords

  • Activity-based probes
  • Breast cancer
  • Cysteine cathepsins
  • Premalignant progression
  • Protease inhibitors

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