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Am J Physiol Cell Physiol 279: C709-C716, 2000;
0363-6143/00 $5.00
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Vol. 279, Issue 3, C709-C716, September 2000

Impairment of NF-kappa B activation and modulation of gene expression by calpastatin

Fei Chen1, Laurence M. Demers2, Val Vallyathan1, Yongju Lu1, Vincent Castranova1, and Xianglin Shi1

1 Pathology and Physiology Research Branch, Health Effects Laboratory Division, National Institute for Occupational Safety and Health, Morgantown, West Virginia 26505; and 2 Department of Pathology, Pennsylvania State University College of Medicine, Hershey, Pennsylvania 17033

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 (Ikappa Balpha ) 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. Genefilter 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.

nuclear factor-kappa B; calpain; silica; epithelial cells


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