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首页> 外文期刊>Biological chemistry >The Calpastatin-Derived Calpain Inhibitor CP1B Reduces mRNA Expression of Matrix Metalloproteinase-2 and -9 and Invasion by Leukemic THP-1 Cells
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The Calpastatin-Derived Calpain Inhibitor CP1B Reduces mRNA Expression of Matrix Metalloproteinase-2 and -9 and Invasion by Leukemic THP-1 Cells

机译：钙抑素衍生的钙蛋白酶抑制剂CP1B降低基质金属蛋白酶-2和-9的mRNA表达以及白血病THP-1细胞的侵袭

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The ubiquitous proteases μ- and m-calpain are Ca~(2+)-dependent cysteine endopeptidases. Besides involvement in a variety of physio (patho) ogical processes, recent studies suggest a pivotal role of calpains in differentiation of hematopoietic cells and tumor cell invasion. However, the precise actions of calpains and their endogenous inhibitor, calpastatin, in these processes are only partially understood. Here we have studied the role of the calpain/calpastatin system in the invasion of leukemic cells under basal and diferentiation-stimulating conditions. To further differentiate the human leukaemic cell line THP-1 (monocytic), the cells were treated for 24 hours with the differentiation-stimulating reagents phorbol 12-myristate 13-acetate (PMA) and dimethyl sulfoxide (DMSO). Macrophage- and granulocyte-like differentiation was confirmed by induction of vimentin expression as well as by microscopic and fluorescence-assisted cytometric analysis. Extracellular matrix (ECM) invasion of both the basal and differentiation-stimulated cells in a Matrigel assay was inhibited by pre-incubation of the cells with the specific calpain inhibitor CP1B for 24 hours. Inhibition of invasiveness correlated with decreased mRNA expression and secretion of the matrix metallopoteinases MMP-2 and MMP-9. In contrast, addition of CP1B only during the invasion process did neither influence transmigration nor MMP release. This is the first report showing that the calpain/calpastatin system mediates MMP-mRNA expression of the leukemic THP-1 cells and as a consequence their invasiveness.
机译：普遍存在的蛋白酶μ-和m-钙蛋白酶是Ca〜（2+）依赖的半胱氨酸内肽酶。除了参与各种生理（病理）过程外，最近的研究表明钙蛋白酶在造血细胞分化和肿瘤细胞侵袭中起着关键作用。但是，钙蛋白酶及其内源性抑制剂钙蛋白酶抑制素在这些过程中的确切作用仅得到部分了解。在这里，我们研究了钙蛋白酶/钙蛋白酶抑制剂系统在基础和分化刺激条件下在白血病细胞侵袭中的作用。为了进一步分化人白血病细胞系THP-1（单核细胞），用分化刺激试剂佛波12-肉豆蔻酸酯13-乙酸酯（PMA）和二甲基亚砜（DMSO）处理细胞24小时。巨噬细胞和粒细胞样分化已通过波形蛋白表达的诱导以及显微镜和荧光辅助细胞计数分析得到证实。通过将细胞与特定钙蛋白酶抑制剂CP1B预温育24小时，可以抑制Matrigel分析中基底细胞和分化刺激细胞的细胞外基质（ECM）入侵。侵袭性的抑制与基质金属蛋白酶MMP-2和MMP-9的mRNA表达降低和分泌有关。相反，仅在入侵过程中添加CP1B既不影响转运也不影响MMP释放。这是第一个报告，表明钙蛋白酶/钙蛋白酶抑制剂系统介导白血病THP-1细胞的MMP-mRNA表达，并因此介导其侵袭性。

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