| dc.description.abstract | Background: E-cadherin is an important adhesion protein that plays a significant role in cell-cell adhesion and tissue integrity in epithelial tissues. It regulates migration and signaling pathways, as well as stabilize cell layers by forming adherens junctions. Genetic modification that alters the expression of E-cadherin can led to changes in cell adhesion and behavior. This study aimed to investigate the role of E-cadherin knockout cells in collective cell migration in keratinocyte monolayers.
Method: The methodology includes the use of CRISPR-Cas9 to create E-cadherin knockout cell clones. Gene expression was assessed through PCR genotyping and Western blotting, and protein localization and expression were analyzed using Immunofluorescence and Traction Force Microscopy. With HaCaT wild-type as a control test to compare the changes in cell adhesion, cytoskeletal organization, and traction forces. In addition, live cell imagining was performed to observe collective cell migration and morphology.
Results: E-cadherin knockout clones maintained the ability to form confluent monolayers similar to HaCaT wild-type cells, as well as the patterns of topological defects in wild-type was also founded in the clones. This could indicate that other adhesion mechanisms compensate for the loss of E-cadherin. Traction Force Microscopy display that E-cadherin knockout clones has increasing traction force and cell-cell tension compared to wild-type cells. The analysis of Immunofluorescence also showed altered expression of focal adhesion proteins like paxillin and vinculin. Interestingly, tight junctions and cytoskeletal integrity remained mostly intact even the loss of E-cadherin.
Conclusion: While E-cadherin is essential for maintain cellular organization during migration, the findings has showed that epithelial tissue can adapt to the loss of E-cadherin. This suggest that other adhesion proteins and compensatory pathways may have taken over the process. | |
