RhoA and Rac1 in Liver Cancer Cells: Induction of Overexpression Using Mechanical Stimulation
Abstract
:1. Introduction
2. Materials and Methods
2.1. An Electromagnetic Actuation Device for Cell Stretching
2.2. Seeding and Culturing Liver Cancer Cells on the Deformable Membrane of the Electromagnetic Device
2.3. Applying Cyclic Stretching on the Cultured Cells
2.4. Cell Lysis Procedure
2.5. Quantification and Detection of RhoA and Rac1 Biomarkers Using ELISA Technique
2.6. Statistical Analysis
2.7. Immunofluorosence Staining
2.8. Fluorescence Microscopy
3. Results and Discussion
3.1. Qualitative Analysis of Effect of Stretching on Liver Cancer Cells
3.2. Quantitative Analysis of Effect of Stretching on Liver Cancer Cells
3.2.1. Difference between the Concentrations of RhoA and Rac1 before and after Applying Mechanical Strain
3.2.2. Time-Dependent Expression of RhoA
3.2.3. Detection of RhoA in Multiple Cell Lines
3.2.4. Time-Dependent Expression of Rac1
3.2.5. Detection of Rac1 in Multiple Cell Lines
3.2.6. Time-Dependent Activation of AFP
4. Conclusions
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Appendix A
Appendix A.1. Different Cell Lysis Methods
Appendix A.2. Standard Curve
Appendix A.3. Quantification of Control Marker AFP
S.N. | Comparison | p Value | Summary |
---|---|---|---|
1 | PLC/PRF/5 No stretch vs. 2 h stretch No stretch vs. 4 h stretch | 0.5139 0.8373 | ns ns |
2 | Hep G2 No stretch vs. 2 h stretch No stretch vs. 4 h stretch | 0.9045 >0.9999 | ns ns |
3 | Hep 3B No stretch vs. 2 h stretch No stretch vs. 4 h stretch | 0.9033 0.8183 | ns ns |
4 | SNU 245 No stretch vs. 2 h stretch No stretch vs. 4 h stretch | 0.5188 0.8486 | ns ns |
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Yadav, S.; Kashaninejad, N.; Nguyen, N.-T. RhoA and Rac1 in Liver Cancer Cells: Induction of Overexpression Using Mechanical Stimulation. Micromachines 2020, 11, 729. https://doi.org/10.3390/mi11080729
Yadav S, Kashaninejad N, Nguyen N-T. RhoA and Rac1 in Liver Cancer Cells: Induction of Overexpression Using Mechanical Stimulation. Micromachines. 2020; 11(8):729. https://doi.org/10.3390/mi11080729
Chicago/Turabian StyleYadav, Sharda, Navid Kashaninejad, and Nam-Trung Nguyen. 2020. "RhoA and Rac1 in Liver Cancer Cells: Induction of Overexpression Using Mechanical Stimulation" Micromachines 11, no. 8: 729. https://doi.org/10.3390/mi11080729