Calmodulin downregulation in conditional knockout HeLa cells inhibits cell migration.
Arch Biochem Biophys
; 697: 108680, 2021 01 15.
Article
en En
| MEDLINE
| ID: mdl-33220265
ABSTRACT
The study of calmodulin (CaM) functions in living cells has been tackled up to date using cell-permeant CaM inhibitors or interference-RNA methods. CaM inhibitors may lack specificity and the siRNA interference approach is challenging, as all three CaM genes expressing an identical protein in mammals have to be blocked. Therefore, we recently introduced a novel genetic system using CRISPR/Cas9-mediated gene deletion and conditional CaM expression to study the function of CaM in HeLa cells. Here, we describe the effect of CaM downregulation on the basal and epidermal growth factor (EGF)-dependent 2D- and 3D-migration in HeLa cells. CaM downregulation inhibited cell migration on a 2D-surface in the absence but not in the presence of EGF. In contrast, CaM downregulation led to inhibition of 3D-migration across a porous membrane both in the absence and presence of EGF. CaM downregulation decreased the expression of Rac1, Cdc42 and RhoA, all known to play crucial roles in cell migration. These results show that EGF-dependent 2D- and 3D-migration utilize distinct CaM-regulated systems and identify several essential migratory proteins directly or indirectly regulated by CaM.
Palabras clave
Texto completo:
1
Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
Calmodulina
/
Regulación hacia Abajo
/
Movimiento Celular
/
Técnicas de Inactivación de Genes
Tipo de estudio:
Prognostic_studies
Límite:
Humans
Idioma:
En
Revista:
Arch Biochem Biophys
Año:
2021
Tipo del documento:
Article
País de afiliación:
España