MIGRATION OF 3T3 AND LUNG FIBROBLASTS IN RESPONSE TO CALCITONIN GENE-RELATED PEPTIDE AND BOMBESIN

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Abstract
Neuropeptides found in airways, such as bombesin and calcitonin gene-related peptide (CGRP), are known to elicit proliferation of several cell types, including fibroblasts and epithelial cells in culture and in vivo. However, the effects of these neuropeptides on fibroblast migration, which may also be a necessary part of airway repair, have not been well established. To determine if these peptides could stimulate fibroblast chemotaxis, we grew NIH 3T3 and IMR-90 cells in culture and studied migration using 48-well blindwell chambers. 3T3 cells were treated with 1014 to 107M bombesin or 1014 to 107M CGRP and permitted to migrate through a gelatin-coated filter for 2-24 hours. Both bombesin and CGRP elicited 3T3 migration which was both time and concentrationdependent. After 6 hours, migration of 3T3 cells treated with 107M bombesin was 33.9 +/- 4.4 cells versus control migration of 4.0 +/- 1.2 cells per 10 high-power fields (hpf) (P<.01, n = 4). Migration of 3T3 cells treated with 109M CGRP was 30.2 +/- 5.4 cells versus control migration of 10.7 +/- 1.4 cells per 10 hpf (P<0.02, n = 6). IMR-90 cells migrated in a similar manner in response to CGRP and bombesin. The response to each neuropeptide was both chemotactic and chemokinetic, and could be blocked completely with an appropriate receptor antagonist. We demonstrate that both bombesin and CGRP are chemotactic for 3T3 and IMR-90 fibroblasts in culture. These peptides therefore may have multiple roles in repair and healing of airway injury.

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