The 1996 Lindberg Award. Calcium antagonists alter cell shape and induce procollagenase synthesis in keloid and normal human dermal fibroblasts

Abstract

Fibroblast cytomorphology is tightly coupled to phenotypic expression, particularly as it relates to extracellular matrix protein synthesis and degradation. We have observed that calcium antagonists, such as verapamil and trifluoperazine, depolymerize actin filaments and alter fibroblast cell shape from bipolar to spherical. Characteristically, the depolymerization of actin filaments, which mediates the cell shape change, turns on procollagenase gene expression in normal human skin fibroblasts. We have found the same effects of calcium antagonists on cell shape, cytoskeletal components, and induction of procollagenase in the keloid fibroblasts of three cell lines, CB792, CW792, and WT949. Rounded cells were seen in 74.8% of verapamil-treated and 86.7% of trifluoperazine-treated cells, whereas only 1.1% of the control cells were spherical. The percentage of cells that synthesized collagenase in the control, verapamil-treated, and trifluoperazine-treated groups was 3.8%, 42.8%, and 53.4%, respectively. Approximately 60% of rounded cells exhibited increased collagenase synthesis when the cells were treated with a calcium antagonist. These results indicate considerable heterogeneity in the phenotypic response to morphologic change. The amount of procollagenase synthesized in a cell was estimated by the fluorescence intensity of the fluorescein-labeled antibody. The normalized fluorescence intensity of procollagenase in the control cells was about 2 to 2.6 times that of background. In contrast, the normalized fluorescence intensity of procollagenase in the calcium antagonist-treated cells was about 2.4 to 12 times that of background. This high intensity level indicates an increase in procollagenase production in the calcium antagonist-treated cells. Calcium green dye used to study cytosolic calcium revealed that after cells were treated with verapamil, the cytosolic calcium ion concentration first increased and then decreased. The change of cytosolic calcium ion concentration may be related to the depolymerization of actin filaments and the alteration of cell shape.