Impairment of NF-κB activation and modulation of gene expression by calpastatin
Impairment of NF-κB activation and modulation of gene expression by calpastatin
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DOI：
10.1152/ajpcell.2000.279.3.c709
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发表时间：
2000-09-01
影响因子：
5.5
通讯作者：
Shi, XL
中科院分区：
文献类型：
--
作者：
Chen, F;Demers, LM;Shi, XL
为了解决钙蛋白酶系统参与基础和二氧化硅诱导的核因子（NF）-κ B B激活，建立了几个人支气管上皮细胞系，其中钙蛋白酶的细胞内抑制剂，钙蛋白酶抑制剂，稳定表达。在稳定过表达钙蛋白酶抑制蛋白的细胞中观察到基础和二氧化硅诱导的抑制剂（I κ B α）降解和NF-κ B活化减少。此外，其中钙蛋白酶被钙蛋白酶抑制素的过表达组成性抑制的细胞表现出显着的形态学变化。而空载体转染的细胞显示出与亲本细胞的形态学无法区分，过表达钙蛋白酶抑制蛋白的细胞表现出马赛克形态学变化，细胞接种后30分钟形成的板层减少。基因过滤器微阵列实验，其中3,965个人类基因可以同时评估其表达，表明钙蛋白酶抑制蛋白下调编码几种膜相关蛋白或核蛋白的基因，上调胶原蛋白α 2，DAZ和线粒体囊硒蛋白的基因。这些结果表明，除了对细胞骨架蛋白和其他细胞调节蛋白的蛋白水解活性外，钙蛋白酶-钙蛋白酶抑制蛋白系统还可以影响编码结构蛋白或调节蛋白的基因的表达水平。
To address the involvement of the calpain system in both basal and silica-induced nuclear factor (NF)-kappa B activation, several human bronchial epithelial cell lines were established in which an intracellular inhibitor of calpain, calpastatin, was stably expressed. Reduced basal and silica-induced inhibitor (I kappa B alpha) degradation and NF-kappa B activation were observed in cells stably overexpressing calpastatin. In addition, the cells in which calpain was constitutively inhibited by the overexpression of calpastatin exhibited a notable morphological change. Whereas empty vector-transfected cells displayed a morphology indistinguishable from that of parental cells, cells overexpressing calpastatin exhibited a mosaic morphological change with reduced formation of lamella 30 min after the cells were seeded. Gene-filter microarray experiments, in which 3,965 human genes can be evaluated for their expression at the same time, showed that calpastatin downregulated genes encoding several membrane-associated proteins or nuclear proteins and upregulated genes of collagen alpha 2, DAZ, and mitochondrial capsule selenoprotein. These results suggest that, in addition to their proteolytic activities on cytoskeletal proteins and other cellular regulatory proteins, calpain-calpastatin systems can also affect the expression levels of genes encoding structural or regulatory proteins.