Protease activity at invadopodial focal digestive areas is dependent on NHE1-driven acidic pHe

Greco,Maria Raffaella; Antelmi,Ester; Busco,Giovanni; Guerra,Lorenzo; Rubino,Rosa; Casavola,Valeria; Reshkin,Stephan Joel; Cardone,Rosa Angela

doi:10.3892/or.2013.2923

Protease activity at invadopodial focal digestive areas is dependent on NHE1-driven acidic pHe

Authors:
- Maria Raffaella Greco
- Ester Antelmi
- Giovanni Busco
- Lorenzo Guerra
- Rosa Rubino
- Valeria Casavola
- Stephan Joel Reshkin
- Rosa Angela Cardone
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Affiliations: Department of Biosciences, Biotechnology and Biopharmaceutics, University of Bari, I-70126 Bari, Italy
Published online on: December 13, 2013 https://doi.org/10.3892/or.2013.2923
Pages: 940-946

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Abstract

Degradation of the extracellular matrix (ECM) is a critical step of tumor cell invasion and requires protease-dependent proteolysis focalized at the invadopodia where the proteolysis of the ECM occurs. Most of the extracellular proteases belong to serine- or metallo-proteases and the invadopodia is where protease activity is regulated. While recent data looking at global protease activity in the growth medium reported that their activity and role in invasion is dependent on Na+/H+ exchanger 1 (NHE1)-driven extracellular acidification, there is no data on this aspect at the invadopodia, and an open question remains whether this acid extracellular pH (pHe) activation of proteases in tumor cells occurs preferentially at invadopodia. We previously reported that the NHE1 is expressed in breast cancer invadopodia and that the NHE1‑dependent acidification of the peri-invadopodial space is critical for ECM proteolysis. In the present study, using, for the first time, in situ zymography analysis, we demonstrated a concordance between NHE1 activity, extracellular acidification and protease activity at invadopodia to finely regulate ECM digestion. We demonstrated that: (i) ECM proteolysis taking place at invadopodia is driven by acidification of the peri-invadopodia microenvironment; (ii) that the proteases have a functional pHe optimum that is acidic; (iii) more than one protease is functioning to digest the ECM at these invadopodial sites of ECM proteolysis; and (iv) lowering pHe or inhibiting the NHE1 increases protease secretion while blocking protease activity changes NHE1 expression at the invadopodia.

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