A recombinant matriptase causes an increase in caspase-3 activity in a small-intestinal epithelial IEC-6 line cultured on fibronectin-coated plates
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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重组基质酶导致在纤连蛋白包被板上培养的小肠上皮 IEC-6 系中 caspase-3 活性增加
DOI：
10.1007/s10616-013-9582-2
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发表时间：
2014
期刊：
影响因子：
2.2
通讯作者：
Kuniyo Inouye & Tohru Fushiki
中科院分区：
文献类型：
--
作者：
Seiya Mochida ;Satoshi Tsuzuki ;Kuniyo Inouye & Tohru Fushiki
间质蛋白酶是上皮来源的II型跨膜丝氨酸蛋白酶。这种蛋白酶在小肠上皮细胞的绒毛尖端显著表达，在该处衰老细胞经历脱落和/或凋亡。上皮细胞（包括小肠上皮细胞）的基底膜含有细胞外基质（ECM）蛋白，例如纤连蛋白和层粘连蛋白。我们以前发现，高浓度的重组间质蛋白酶催化结构域（r-MatCD）（例如1 μM）引起在层粘连蛋白包被的平板上培养的大鼠小肠上皮IEC-6系中凋亡效应物半胱天冬酶-3的分离增加和活性增加，并提出在其高水平表达的位点，间质蛋白酶通过介导细胞-ECM粘附丧失的机制促进上皮细胞脱落和/或脱落诱导的死亡。在这项研究中，我们发现，即使没有增加细胞分离，高浓度的r-MatCD导致在IEC-6细胞中培养的纤连蛋白包被的板上的caspase-3活性增加，这表明重组matriptase可以通过与细胞分离无关的机制引起细胞凋亡。此外，发现r-MatCD处理的IEC-6细胞在纤连蛋白上显示梭形形态学变化。我们认为r-MatCD通过破坏细胞完整性的机制导致纤连蛋白上的IEC-6细胞凋亡。
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.