The DNA-Damage Response to γ-Radiation Is Affected by miR-27a in A549 Cells
Abstract
:1. Introduction
2. Results and Discussion
2.1. miR-27a Interacts with 3′UTR of ATM Gene
2.2. miR-27a Affects ATM Expression Levels in A549 Cells
2.3. Effects of miR-27a on Proliferation of A549 Cells
2.4. Effects of miR-27a Up-Regulation on Radiosensitivity of A549 Cells
3. Experimental Section
3.1. Cell Culture
3.2. Construction of Recombinant Vectors and Site-Directed Mutagenesis
- ATM Fwd-5′-ATCTAGGAGCTCAGGAGTGGAAGAAGGCACTG-3′
- ATM Rev-5′-ATCTAGTCTAGAACGCTGTCCAAAGTTTTTCC-3′
- ATMdel1 Fwd-5′-AGTGGAAGAAGGCACTCTCAGTGTTGGTGGAC-3′
- ATMdel1 Rev-5′-GTCCACCAACACTGAGAGTGCCTTCTTCCACT-3′
- ATMdel2 Fwd-5′-CAAGGACAAATGAGGAGTAGTTAGATGAAAATATTAATCATAGAATAGTTGTT-3′
- ATMdel2 Rev-5′-AACAACTATTCTATGATTAATATTTTCATCTAACTACTCCTCATTTGTCCTTG-3′
3.3. Transient Transfection and Cell Irradiation
3.4. Total RNA Isolation and qRT-PCR
3.5. Luciferase Reporter Assays
3.6. Clonogenic Survival Assay
3.7. Cell Cycle Analysis
3.8. Immunofluorescence Staining
3.9. Statistical Analysis
4. Conclusions
Acknowledgments
Conflicts of Interest
References
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Di Francesco, A.; De Pittà, C.; Moret, F.; Barbieri, V.; Celotti, L.; Mognato, M. The DNA-Damage Response to γ-Radiation Is Affected by miR-27a in A549 Cells. Int. J. Mol. Sci. 2013, 14, 17881-17896. https://doi.org/10.3390/ijms140917881
Di Francesco A, De Pittà C, Moret F, Barbieri V, Celotti L, Mognato M. The DNA-Damage Response to γ-Radiation Is Affected by miR-27a in A549 Cells. International Journal of Molecular Sciences. 2013; 14(9):17881-17896. https://doi.org/10.3390/ijms140917881
Chicago/Turabian StyleDi Francesco, Andrea, Cristiano De Pittà, Francesca Moret, Vito Barbieri, Lucia Celotti, and Maddalena Mognato. 2013. "The DNA-Damage Response to γ-Radiation Is Affected by miR-27a in A549 Cells" International Journal of Molecular Sciences 14, no. 9: 17881-17896. https://doi.org/10.3390/ijms140917881