Cerium Oxide Nanoparticles Regulate Oxidative Stress in HeLa Cells by Increasing the Aquaporin-Mediated Hydrogen Peroxide Permeability
Abstract
:1. Introduction
2. Results
2.1. Aquaporins mRNA and Protein Expression in HeLa Cells
2.2. CNPs and Aquaporin-3, -6, -8, and -11 Localization in HeLa Cells
2.3. The Effect of CNPs on Water Permeability of HeLa Cells in Normal and Oxidative Stress Conditions
2.4. Effect of CNPs in HeLa Cells with Reduced Expression of Single AQPs
3. Discussion
4. Materials and Methods
4.1. Cell Culture
4.2. Cerium Nanoparticles Preparation and Characterization
4.3. RNA Isolation and RT-qPCR
4.4. SDS-PAGE and Immunoblotting
4.5. Double Immunofluorescence
4.6. Gene Silencing
4.7. Water Permeability Measurements
4.8. Hydrogen Peroxide Indicator Transfection for Optical Imaging
4.9. Intracellular H2O2 Detection by HyPer7-NES Imaging
4.10. Protein Content
4.11. Statistics
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Gene | Primer Sequences | Size (bp) | Accession Number | |
---|---|---|---|---|
AQP1 a | Forward | 5’-ACACCTCCTGGCTATTGACTACAC-3’ | 134 | NM_198098; variants 1, 5 |
Reverse | 5’-CCGATGAATGGCCCCACCCAGAA-3’ | |||
AQP2 a | Forward | 5’-CACCTCCTTGGGATCCATTACACC-3’ | 95 | NM_000486 |
Reverse | 5’-ACCCAGTGGTCATCAAATTTGCC-3’ | |||
AQP3 b | Forward | 5’-CCTGGTGATGTTTGGCTGTGGCTC-3’ | 147 | NM_004925; variants 1, 2 |
Reverse | 5’-TTCAGGTGGGCCCCAGAGACC-3’ | |||
AQP4 | Forward | 5’-GGAGTCACCATGGTTCATGGAA-3’ | 123 | NM_001650; variants 1–3 |
Reverse | 5’-AGTGACATCAGTCCGTTTGGAA-3’ | |||
AQP5 | Forward | 5’-GGTGGTGGAGCTGATTCTGA-3’ | 142 | NM_001651 |
Reverse | 5’-GAAGTAGATTCCGACAAGGTGG-3’ | |||
AQP6 | Forward | 5’-CACCTCATTGGGATCCACTTC-3’ | 103 | NM_ 001652; variants 1, 2 |
Reverse | 5’-CCCAGAAGACCCAGTGGACT-3’ | |||
AQP7 | Forward | 5’-GGACAGCTGATGGTGACCGG-3’ | 104 | NM_001170; variants 1–4 |
Reverse | 5’-AGCCACGCCTCATTCAGGAA-3’ | |||
AQP8 | Forward | 5’-TGGAGAGATAGCCATGTGTGAG-3’ | 106 | NM_001169 |
Reverse | 5’-TGGCTGCACAAACCGTTCGT-3’ | |||
AQP9 | Forward | 5’-CCCAGCTGTGTCTTTAGCAA-3’ | 133 | NM_020980; variants 1–3 |
Reverse | 5’-AAGTCCATCATAGTAAATGCCAAA-3’ | |||
AQP10 | Forward | 5’-CCTATGTTCTCTACCATGATGCCC-3’ | 137 | NM_080429 |
Reverse | 5’-CTGATCCAGGAAGCCATTGTTC-3’ | |||
AQP11 | Forward | 5’-TTTCTCTTCCACAGCGCTCT-3’ | 115 | NM_173039; variant 1 |
Reverse | 5’-CTCCTGTTAGACTTCCTCCTGC-3’ | |||
B2M | Hs_B2M_1_SG QuantiTect Primer Assay QT00088935, Qiagen | 98 | NM_004048 |
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Pellavio, G.; Sommi, P.; Anselmi-Tamburini, U.; DeMichelis, M.P.; Coniglio, S.; Laforenza, U. Cerium Oxide Nanoparticles Regulate Oxidative Stress in HeLa Cells by Increasing the Aquaporin-Mediated Hydrogen Peroxide Permeability. Int. J. Mol. Sci. 2022, 23, 10837. https://doi.org/10.3390/ijms231810837
Pellavio G, Sommi P, Anselmi-Tamburini U, DeMichelis MP, Coniglio S, Laforenza U. Cerium Oxide Nanoparticles Regulate Oxidative Stress in HeLa Cells by Increasing the Aquaporin-Mediated Hydrogen Peroxide Permeability. International Journal of Molecular Sciences. 2022; 23(18):10837. https://doi.org/10.3390/ijms231810837
Chicago/Turabian StylePellavio, Giorgia, Patrizia Sommi, Umberto Anselmi-Tamburini, Maria Paola DeMichelis, Stefania Coniglio, and Umberto Laforenza. 2022. "Cerium Oxide Nanoparticles Regulate Oxidative Stress in HeLa Cells by Increasing the Aquaporin-Mediated Hydrogen Peroxide Permeability" International Journal of Molecular Sciences 23, no. 18: 10837. https://doi.org/10.3390/ijms231810837