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Calmodulin downregulation in conditional knockout HeLa cells inhibits cell migration

Juan Alcalde1, Mads Munk2, María González-Muñoz3

  • 1Department of Biology, University of Copenhagen, 13 Universitetsparken, DK-2100, Copenhagen Ø, Denmark; Instituto de Investigaciones Biomédicas, Consejo Superior de Investigaciones Científicas and Universidad Autónoma de Madrid, Madrid, Spain.

Archives of Biochemistry and Biophysics
|November 21, 2020
PubMed

Insights

Calmodulin (CaM) downregulation impacts cell migration differently on 2D and 3D surfaces, affecting basal and epidermal growth factor (EGF)-dependent movement. This study reveals distinct CaM-regulated systems in cell migration.

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biochemistry

Background:

  • Studying calmodulin (CaM) function in cells traditionally uses inhibitors or RNA interference, which have limitations.
  • CaM inhibitors may lack specificity, and blocking all three mammalian CaM genes is challenging.
  • A novel CRISPR/Cas9 system enables precise CaM gene deletion and conditional expression for functional studies.

Purpose of the Study:

  • To investigate the role of calmodulin (CaM) in regulating cell migration.
  • To differentiate the effects of CaM downregulation on 2D versus 3D cell migration.
  • To explore CaM's influence on epidermal growth factor (EGF)-dependent migration.

Main Methods:

  • Utilized CRISPR/Cas9 gene editing for conditional CaM expression and gene deletion in HeLa cells.
  • Assessed cell migration in 2D and 3D environments under basal and EGF-stimulated conditions.
  • Quantified the expression levels of key migration-related proteins (Rac1, Cdc42, RhoA).

Main Results:

  • CaM downregulation inhibited basal 2D cell migration but not EGF-stimulated 2D migration.
  • CaM downregulation inhibited 3D cell migration in both basal and EGF-stimulated conditions.
  • Reduced expression of Rac1, Cdc42, and RhoA was observed upon CaM downregulation.

Conclusions:

  • Epidermal growth factor (EGF)-dependent 2D and 3D cell migration involve distinct calmodulin (CaM)-regulated pathways.
  • Calmodulin (CaM) plays a critical role in cell migration, potentially by regulating proteins like Rac1, Cdc42, and RhoA.
Keywords:
CalmodulinCdc42Cell migrationConditional knock-out cellsRac1RhoA

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