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Electroporation and carrier DNA cause p53 activation, cell cycle arrest, and apoptosis
Dina Lepik1, Viljar Jaks, Lilian Kadaja
1Department of Cell Biology, Institute of Molecular and Cell Biology, Tartu University, 23 Riia Street, Tartu 51010, Estonia. dlepik@ebc.ee
Abstract:
Methods used in transient transfection of cells may alter cellular signaling pathways that in turn may lead to misinterpretation of the results. A variety of genotoxic agents cause the accumulation of the p53 protein leading to either apoptosis or growth arrest. Here we report the effect of electroporation and carrier DNA on the stability, cellular localization, and transcriptional activity of p53. We show that electroporation leads to p53-dependent and also p53-independent cell-cycle arrest and apoptosis. At the same time a chemical agent polyethylenimine that is also used for transient transfection of cells causes neither upregulation of p53 nor cellular response.
Insights
Electroporation causes cell cycle arrest and apoptosis, affecting p53 protein stability and activity. Polyethylenimine, a transfection agent, did not impact p53 or cellular response.
Area of Science:
- Cell Biology
- Molecular Biology
- Biotechnology
Background:
- Transient transfection methods can alter cellular signaling pathways, potentially leading to inaccurate experimental results.
- The p53 protein plays a critical role in cellular responses to genotoxic stress, mediating apoptosis or growth arrest.
Purpose of the Study:
- To investigate the impact of electroporation and carrier DNA on the stability, cellular localization, and transcriptional activity of the p53 protein.
- To compare the cellular effects of electroporation with those of polyethylenimine, another common transfection agent.
Main Methods:
- Cells were subjected to electroporation and transfection with carrier DNA.
- The stability, cellular localization, and transcriptional activity of p53 were analyzed.
- Cellular responses, including cell-cycle arrest and apoptosis, were assessed.
- The effects of polyethylenimine on p53 and cellular responses were also evaluated.
Main Results:
- Electroporation induced both p53-dependent and p53-independent cell-cycle arrest and apoptosis.
- Carrier DNA did not significantly alter these effects.
- Polyethylenimine did not lead to p53 upregulation or any observable cellular response.
Conclusions:
- Electroporation is a potent method that can trigger significant cellular responses, including apoptosis and cell-cycle arrest, independent of p53 in some cases.
- The choice of transfection method is critical, as electroporation significantly impacts cellular pathways, while polyethylenimine appears to have a minimal effect on p53-mediated responses.