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A recombinant matriptase causes an increase in caspase-3 activity in a small-intestinal epithelial IEC-6 line cultured on fibronectin-coated plates

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Abstract

Matriptase is an epithelial-derived type-II transmembrane serine protease. This protease is expressed prominently in the villus tip of small-intestinal epithelia at which senescent cells undergo shedding and/or apoptosis. The basement membrane of epithelial cells, including small-intestinal epithelial cells, contains extracellular matrix (ECM) proteins such as fibronectin and laminin. We found previously that high concentrations of a recombinant matriptase catalytic domain (r-MatCD) (e.g. 1 μM) caused an increased detachment of and increases in the activity of apoptotic effector caspase-3 in a rat small-intestinal epithelial IEC-6 line cultured on laminin-coated plates and proposed that at sites with its high level of expression, matriptase contributes to promoting shedding and/or detachment-induced death of epithelial cells through a mechanism mediating loss of cell-ECM adhesion. In this study, we found that even without increasing cell detachment, a high concentration of r-MatCD causes an increase in caspase-3 activity in IEC-6 cells cultured on fibronectin-coated plates, suggesting that the recombinant matriptase can cause apoptosis by a mechanism unrelated to cell detachment. Also, r-MatCD-treated IEC-6 cells on fibronectin were found to display spindle-like morphological changes. We suggest that r-MatCD causes apoptosis of IEC-6 on fibronectin by a mechanism involving the disruption of cell integrity.

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Abbreviations

Ac:

Acetyl

ADAM:

A disintegrin and metalloprotease

DMEM:

Dulbecco’s modified Eagle’s medium

ECM:

Extracellular matrix

EMT:

Epithelial-to-mesenchymal transition

MMP:

Matrix metalloprotease

SFM:

Serum-free DMEM supplemented with insulin and bovine serum albumin

MCA:

4-Methylcoumaryl-7-amide

r-EK:

Recombinant form of enterokinase catalytic domain

SIECs:

Small-intestinal epithelial cells

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Acknowledgments

This work was supported in part by Grants-in-Aid for Scientific Research from Japan Society for the Promotion of Science.

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Authors and Affiliations

  1. Laboratory of Nutrition, Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Sakyo-ku, Kyoto, 606-8502, Japan

    Seiya Mochida, Satoshi Tsuzuki & Tohru Fushiki

  2. Laboratory of Enzyme Chemistry, Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Sakyo-ku, Kyoto, 606-8502, Japan

    Kuniyo Inouye

Authors
  1. Seiya Mochida
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  2. Satoshi Tsuzuki
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  3. Kuniyo Inouye
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  4. Tohru Fushiki
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Correspondence to Satoshi Tsuzuki.

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Mochida, S., Tsuzuki, S., Inouye, K. et al. A recombinant matriptase causes an increase in caspase-3 activity in a small-intestinal epithelial IEC-6 line cultured on fibronectin-coated plates. Cytotechnology 66, 357–363 (2014). https://doi.org/10.1007/s10616-013-9582-2

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