RhoA-Dependent HGF and c-Met Mediate Gas6-Induced Inhibition of Epithelial–Mesenchymal Transition, Migration, and Invasion of Lung Alveolar Epithelial Cells
Abstract
:1. Introduction
2. Materials and Methods
2.1. Reagents
2.2. Cell Lines and Culture
2.3. Isolation and Culture of Primary Cells
2.4. Incubation of Epithelial Cells
2.5. Transient Transfections
2.6. Immunoblot Analysis
2.7. Quantitative Real-Time Polymerase Chain Reaction (qPCR)
2.8. RhoA Activity Assay
2.9. Enzyme-Linked Immunosorbent Assay (ELISA) Measurement
2.10. Migration and Invasion Assays
2.11. Statistical Analysis
3. Results
3.1. Growth Arrest-Specific Protein 6 (Gas6)/Axl or Mer Signaling Induces RhoA-Dependent Hepatocyte Growth Factor (HGF) Secretion and c-Met Expression in Lung Epithelial Cells
3.2. Axl and Mer Receptor Tyrosine Kinases Mediate Gas6-Induced RhoA Activity, and Expression of HGF and c-Met in LA-4 Cells
3.3. RhoA is Involved in the Gas6-Induced Inhibition of Epithelial–Mesenchymal Transition (EMT) Marker Changes in LA-4 Cells
3.4. RhoA is Involved in the Inhibition of Phospho-Extracellular Signal-Related Kinase (ERK1/2) and AKT Activation and Reduction of EMT-Regulating Transcription Factor Expression by Gas6 in LA-4 Cells
3.5. The Rho Kinase is Involved in the Inhibition of EMT Marker Changes and Reduction of EMT-Regulating Transcription Factor Expression by Gas6 in Lung Epithelial Cells
3.6. c-Met Signaling is Involved in the Gas6-Induced Inhibition of EMT in LA-4 Cells and Primary AT II Cells
3.7. Treatment of Lung Epithelial Cells with Gas6 Inhibits the Migration and Invasion of Epithelial Cells via RhoA-Dependent HGF Secretion
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
References
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Jung, J.; Yang, K.; Kim, H.-J.; Lee, Y.-J.; Kim, M.; Choi, Y.-H.; Kang, J.L. RhoA-Dependent HGF and c-Met Mediate Gas6-Induced Inhibition of Epithelial–Mesenchymal Transition, Migration, and Invasion of Lung Alveolar Epithelial Cells. Biomolecules 2019, 9, 565. https://doi.org/10.3390/biom9100565
Jung J, Yang K, Kim H-J, Lee Y-J, Kim M, Choi Y-H, Kang JL. RhoA-Dependent HGF and c-Met Mediate Gas6-Induced Inhibition of Epithelial–Mesenchymal Transition, Migration, and Invasion of Lung Alveolar Epithelial Cells. Biomolecules. 2019; 9(10):565. https://doi.org/10.3390/biom9100565
Chicago/Turabian StyleJung, Jihye, Kyungwon Yang, Hee-Ja Kim, Ye-Ji Lee, Minsuk Kim, Youn-Hee Choi, and Jihee Lee Kang. 2019. "RhoA-Dependent HGF and c-Met Mediate Gas6-Induced Inhibition of Epithelial–Mesenchymal Transition, Migration, and Invasion of Lung Alveolar Epithelial Cells" Biomolecules 9, no. 10: 565. https://doi.org/10.3390/biom9100565