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Inhibition of endopeptidase and exopeptidase activity of cathepsin B impairs extracellular matrix degradation and tumour invasion

  • Ana Mitrović , Bojana Mirković , Izidor Sosič , Stanislav Gobec and Janko Kos EMAIL logo
Published/Copyright: November 12, 2015

Abstract

Cathepsin B is a lysosomal cysteine protease that is implicated in a number of physiological processes, including protein turnover in lysosomes. Changes in its expression are associated with a variety of pathological processes, including cancer. Due to the structural feature, termed the occluding loop, cathepsin B differs from other cysteine proteases in possessing both, endopeptidase and exopeptidase activity. Here we investigated the impact of both cathepsin B activities on intracellular and extracellular collagen IV degradation and tumour cell invasion using new selective synthetic inhibitors, 2-{[(8-hydroxy-5-nitroquinoline-7-yl)methyl]amino}-acetonitrile (1), 8-(4-methylpiperidin-1-yl)-5-nitroquinoline (2) and 7-[(4-methylpiperidin-1yl)methyl]-5-nitroquinolin-8-ol (3). All three compounds (5 μm) reduced extracellular degradation of collagen IV by MCF-10A neoT cells by 45–70% as determined by spectrofluorimetry and they (50 μm) attenuated intracellular collagen IV degradation by 40-60% as measured with flow cytometry. Furthermore, all three compounds (5 μm) impaired MCF-10A neoT cell invasion by 40–80% as assessed by measuring electrical impedance in real time. Compounds 1 and 3 (5 μm), but not compound 2, significantly reduced the growth of MMTV-PyMT multicellular tumour spheroids. Collectively, these data suggest that the efficient strategy to impair harmful cathepsin B activity in tumour progression may include simultaneous and potent inhibition of cathepsin B endopeptidase and exopeptidase activities.


Corresponding author: Janko Kos, Faculty of Pharmacy, Department of Pharmaceutical Biology, University of Ljubljana, Aškerčeva cesta 7, SI-1000 Ljubljana, Slovenia; and Jožef Stefan Institute, Department of Biotechnology, Jamova cesta 39, SI-1000 Ljubljana, Slovenia, e-mail:
aAna Mitrović and Bojana Mirković: These authors contributed equally to this work.

Acknowledgments

This work was supported by the Slovenian Research Agency (grant number J4-5529 to J.K.).

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Received: 2015-9-1
Accepted: 2015-11-7
Published Online: 2015-11-12
Published in Print: 2016-1-1

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