Beta3-Tubulin Is Critical for Microtubule Dynamics, Cell Cycle Regulation, and Spontaneous Release of Microvesicles in Human Malignant Melanoma Cells (A375)
Abstract
:1. Introduction
2. Results
2.1. A375 Cells Express β3-Tubulin mRNA and Protein
2.2. siRNA Knockdown of β3-Tubulin Reduces the Spontaneous Release of MVs by A375 Cells
2.3. siRNA Knockdown of β3-Tubulin Suppresses MTs Dynamics and Induces G2/M Cell Cycle Arrest
3. Discussion
4. Materials and Methods
4.1. Cell Culture EGFP-MAP-4 Transfection and siRNA Silencing
4.2. RNA Isolation, cDNA Synthesis, and qPCR
4.3. Electrophoresis, Western Analysis, and Fluorescent Microscopy
4.4. MVs Purification and Flowcytometric Quantification
4.5. Analysis of Microtubules Dynamics
4.6. Propidium Iodide (PI)-Staining and Cell Cycle Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
MTs | Microtubules |
MVs | Microvesicles |
WGA | Wheat germ agglutinin |
MAA | Melanoma associated antigen |
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Altonsy, M.O.; Ganguly, A.; Amrein, M.; Surmanowicz, P.; Li, S.S.; Lauzon, G.J.; Mydlarski, P.R. Beta3-Tubulin Is Critical for Microtubule Dynamics, Cell Cycle Regulation, and Spontaneous Release of Microvesicles in Human Malignant Melanoma Cells (A375). Int. J. Mol. Sci. 2020, 21, 1656. https://doi.org/10.3390/ijms21051656
Altonsy MO, Ganguly A, Amrein M, Surmanowicz P, Li SS, Lauzon GJ, Mydlarski PR. Beta3-Tubulin Is Critical for Microtubule Dynamics, Cell Cycle Regulation, and Spontaneous Release of Microvesicles in Human Malignant Melanoma Cells (A375). International Journal of Molecular Sciences. 2020; 21(5):1656. https://doi.org/10.3390/ijms21051656
Chicago/Turabian StyleAltonsy, Mohammed O., Anutosh Ganguly, Matthias Amrein, Philip Surmanowicz, Shu Shun Li, Gilles J. Lauzon, and P. Régine Mydlarski. 2020. "Beta3-Tubulin Is Critical for Microtubule Dynamics, Cell Cycle Regulation, and Spontaneous Release of Microvesicles in Human Malignant Melanoma Cells (A375)" International Journal of Molecular Sciences 21, no. 5: 1656. https://doi.org/10.3390/ijms21051656