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Human plasma can modulate micronucleus frequency in TK6 and OE33 cells in vitro
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Gareth Jenkins
Mutation Research - Genetic Toxicology and Environmental Mutagenesis, Volume: 896, Start page: 503766
Swansea University Authors:
Hamsa Naser, Kathryn Munn, Rhiannon Wright, ETHAN GREWAL, Shareen Doak , Gareth Jenkins
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DOI (Published version): 10.1016/j.mrgentox.2024.503766
Abstract
In this paper, we studied the potential genotoxic effects of human plasma from healthy volunteers, as well as patients with gastro-oesophageal reflux disease, Barrett’s oesophagus (BO) and oesophageal adenocarcinoma (OAC) using the oesophageal adenocarcinoma cell line (OE33) and the lymphoblastoid c...
| Published in: | Mutation Research - Genetic Toxicology and Environmental Mutagenesis |
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| ISSN: | 1383-5718 |
| Published: |
Elsevier BV
2024
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| Online Access: |
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| URI: | https://cronfa.swan.ac.uk/Record/cronfa66474 |
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| Abstract: |
In this paper, we studied the potential genotoxic effects of human plasma from healthy volunteers, as well as patients with gastro-oesophageal reflux disease, Barrett’s oesophagus (BO) and oesophageal adenocarcinoma (OAC) using the oesophageal adenocarcinoma cell line (OE33) and the lymphoblastoid cell line (TK6). Both TK6 and OE33 cells were treated with plasma (10 % volume, replacing foetal bovine serum (FBS) or horse serum (HS)) at different time points of 4 hours (for the micronucleus (Mn) assay and the invasion assay) and 24 hours (for the cell cycle studies). Plasma-induced effects on DNA damage levels, cell viability and the cell cycle were studied by the micronucleus assay, cytokinesis block proliferation index (CBPI) and flow cytometry respectively. The expression of IL-8 in supernatants of TK6 cells and IFN-β in OE33 cells was also analysed by enzyme-linked immunosorbent assay (ELISA). Finally, we carried out an assessment of cellular invasion of OE33 cells following plasma treatment.The results of the micronucleus assay confirmed the genotoxicity of direct plasma treatment from some participants through the increase in DNA damage in TK6 cells. Conversely, some individual patient plasma samples reduced background levels of TK6 cell Mn frequency, in an anti-genotoxic fashion. In TK6 cells, (on average) plasma samples from patients with Barrett’s oesophagus induced higher micronucleus levels than healthy volunteers (p= 0.0019). There was little difference in Mn induction when using plasma versus serum to treat the cells in vitro. Cell cycle results showed that direct plasma treatment had a marked impact on OE33 cells at 24h (p=0.0182 for BO and p=0.0320 for OAC) by decreasing the proportion of cells in the S phase, while plasma exposure was less impactful on the cell cycle of TK6 cells. Invasion of OE33 cells was also seen to be non-significantly affected by plasma treatment of OE33 cells.The addition of N-acetyl cysteine NAC in a dose-dependent matter did not alter the formation of Mn in TK6 cells, suggesting that reactive oxygen species (ROS) are not the root cause of plasma’s genotoxicity. The concentration of IL-8 in TK6 cells and IFN-β in OE33 cells was significantly higher in cells treated with OAC-derived plasma than in the untreated negative control. Collectively, our results demonstrate that plasma-specific effects are detectable which helps us better understand some important aspects of the biology of blood-based biomarkers under development. |
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| Keywords: |
Genotoxicity; Cytotoxicity; DNA damage; micronucleus assay; cell cycle; plasma; serum |
| College: |
Faculty of Medicine, Health and Life Sciences |
| Funders: |
We thank Cancer Research Wales for funding this project and note that a PhD studentship awarded to Kathryn Munn by Swansea University contributed to this work. Dr Hamsa Naser was funded by the Faculty of Medicine, Health and Life Science at Swansea University. |
| Start Page: |
503766 |