In Vitro and In Silico Mechanistic Insights into miR-21-5p-Mediated Topoisomerase Drug Resistance in Human Colorectal Cancer Cells
Abstract
:1. Introduction
2. Materials and Methods
2.1. Chemicals and Reagents
2.2. Cell Culture and Transfection
2.3. Determination of Cell Proliferation and Cell Viability
2.4. Quantitative Real-time Polymerase Chain Reaction (qPCR)
2.5. Western Blot Analysis
2.6. Band-depletion Assay
2.7. Microarray, Connectivity Map (CMap), and Kyoto Encyclopedia of Genes and Genomes (KEGG) Pathway Enrichment Analyses
2.8. CellMinerCDB Analysis
2.9. Kaplan–Meier Survival Analysis
3. Results
3.1. miR-21 Overexpression is Correlated with Drug Resistance to Topoisomerase Inhibitors
3.2. miR-21 Overexpression Attenuates VP-16-Induced Apoptosis without Affecting Expressions or Activities of Topoisomerases
3.3. Accelerated Autophagic flux is Associated with miR-21-Induced Drug Resistance
3.4. In Silico Analyses Reveal the Mechanistic Role of miR-21 Overexpression in the Development of Drug Resistance
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Chen, J.-C.; Hsieh, Y.-Y.; Lo, H.-L.; Li, A.; Chou, C.-J.; Yang, P.-M. In Vitro and In Silico Mechanistic Insights into miR-21-5p-Mediated Topoisomerase Drug Resistance in Human Colorectal Cancer Cells. Biomolecules 2019, 9, 467. https://doi.org/10.3390/biom9090467
Chen J-C, Hsieh Y-Y, Lo H-L, Li A, Chou C-J, Yang P-M. In Vitro and In Silico Mechanistic Insights into miR-21-5p-Mediated Topoisomerase Drug Resistance in Human Colorectal Cancer Cells. Biomolecules. 2019; 9(9):467. https://doi.org/10.3390/biom9090467
Chicago/Turabian StyleChen, Jung-Chien, Yao-Yu Hsieh, Hsiang-Ling Lo, Albert Li, Chia-Jung Chou, and Pei-Ming Yang. 2019. "In Vitro and In Silico Mechanistic Insights into miR-21-5p-Mediated Topoisomerase Drug Resistance in Human Colorectal Cancer Cells" Biomolecules 9, no. 9: 467. https://doi.org/10.3390/biom9090467