Genotoxicity of pyrrolizidine alkaloids in metabolically inactive human cervical cancer HeLa cells co-cultured with human hepatoma HepG2 cells
© 2022. The Author(s)..
Pyrrolizidine alkaloids (PAs) are secondary plant metabolites, which can be found as contaminant in various foods and herbal products. Several PAs can cause hepatotoxicity and liver cancer via damaging hepatic sinusoidal endothelial cells (HSECs) after hepatic metabolization. HSECs themselves do not express the required metabolic enzymes for activation of PAs. Here we applied a co-culture model to mimic the in vivo hepatic environment and to study PA-induced effects on not metabolically active neighbour cells. In this co-culture model, bioactivation of PA was enabled by metabolically capable human hepatoma cells HepG2, which excrete the toxic and mutagenic pyrrole metabolites. The human cervical epithelial HeLa cells tagged with H2B-GFP were utilized as non-metabolically active neighbours because they can be identified easily based on their green fluorescence in the co-culture. The PAs europine, riddelliine and lasiocarpine induced micronuclei in HepG2 cells, and in HeLa H2B-GFP cells co-cultured with HepG2 cells, but not in HeLa H2B-GFP cells cultured alone. Metabolic inhibition of cytochrome P450 enzymes with ketoconazole abrogated micronucleus formation. The efflux transporter inhibitors verapamil and benzbromarone reduced micronucleus formation in the co-culture model. Furthermore, mitotic disturbances as an additional genotoxic mechanism of action were observed in HepG2 cells and in HeLa H2B-GFP cells co-cultured with HepG2 cells, but not in HeLa H2B-GFP cells cultured alone. Overall, we were able to show that PAs were activated by HepG2 cells and the metabolites induced genomic damage in co-cultured HeLa cells.
| Medienart: |
E-Artikel |
|---|
| Erscheinungsjahr: |
2023 |
|---|---|
| Erschienen: |
2023 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:97 |
|---|---|
| Enthalten in: |
Archives of toxicology - 97(2023), 1 vom: 06. Jan., Seite 295-306 |
| Sprache: |
Englisch |
|---|
| Beteiligte Personen: |
Hadi, Naji Said Aboud [VerfasserIn] |
|---|
| Links: |
|---|
| Themen: |
Co-culture |
|---|
| Anmerkungen: |
Date Completed 09.01.2023 Date Revised 18.01.2023 published: Print-Electronic Citation Status MEDLINE |
|---|
| doi: |
10.1007/s00204-022-03394-z |
|---|
| funding: |
|
|---|---|
| Förderinstitution / Projekttitel: |
|
| PPN (Katalog-ID): |
NLM347879535 |
|---|
| LEADER | 01000naa a22002652 4500 | ||
|---|---|---|---|
| 001 | NLM347879535 | ||
| 003 | DE-627 | ||
| 005 | 20231226034901.0 | ||
| 007 | cr uuu---uuuuu | ||
| 008 | 231226s2023 xx |||||o 00| ||eng c | ||
| 024 | 7 | |a 10.1007/s00204-022-03394-z |2 doi | |
| 028 | 5 | 2 | |a pubmed24n1159.xml |
| 035 | |a (DE-627)NLM347879535 | ||
| 035 | |a (NLM)36273350 | ||
| 040 | |a DE-627 |b ger |c DE-627 |e rakwb | ||
| 041 | |a eng | ||
| 100 | 1 | |a Hadi, Naji Said Aboud |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Genotoxicity of pyrrolizidine alkaloids in metabolically inactive human cervical cancer HeLa cells co-cultured with human hepatoma HepG2 cells |
| 264 | 1 | |c 2023 | |
| 336 | |a Text |b txt |2 rdacontent | ||
| 337 | |a ƒaComputermedien |b c |2 rdamedia | ||
| 338 | |a ƒa Online-Ressource |b cr |2 rdacarrier | ||
| 500 | |a Date Completed 09.01.2023 | ||
| 500 | |a Date Revised 18.01.2023 | ||
| 500 | |a published: Print-Electronic | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a © 2022. The Author(s). | ||
| 520 | |a Pyrrolizidine alkaloids (PAs) are secondary plant metabolites, which can be found as contaminant in various foods and herbal products. Several PAs can cause hepatotoxicity and liver cancer via damaging hepatic sinusoidal endothelial cells (HSECs) after hepatic metabolization. HSECs themselves do not express the required metabolic enzymes for activation of PAs. Here we applied a co-culture model to mimic the in vivo hepatic environment and to study PA-induced effects on not metabolically active neighbour cells. In this co-culture model, bioactivation of PA was enabled by metabolically capable human hepatoma cells HepG2, which excrete the toxic and mutagenic pyrrole metabolites. The human cervical epithelial HeLa cells tagged with H2B-GFP were utilized as non-metabolically active neighbours because they can be identified easily based on their green fluorescence in the co-culture. The PAs europine, riddelliine and lasiocarpine induced micronuclei in HepG2 cells, and in HeLa H2B-GFP cells co-cultured with HepG2 cells, but not in HeLa H2B-GFP cells cultured alone. Metabolic inhibition of cytochrome P450 enzymes with ketoconazole abrogated micronucleus formation. The efflux transporter inhibitors verapamil and benzbromarone reduced micronucleus formation in the co-culture model. Furthermore, mitotic disturbances as an additional genotoxic mechanism of action were observed in HepG2 cells and in HeLa H2B-GFP cells co-cultured with HepG2 cells, but not in HeLa H2B-GFP cells cultured alone. Overall, we were able to show that PAs were activated by HepG2 cells and the metabolites induced genomic damage in co-cultured HeLa cells | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Co-culture | |
| 650 | 4 | |a Cytochrome P450s | |
| 650 | 4 | |a Membrane transporters | |
| 650 | 4 | |a Micronuclei | |
| 650 | 4 | |a Mitotic disturbance | |
| 650 | 4 | |a Pyrrolizidine alkaloids | |
| 650 | 7 | |a Pyrrolizidine Alkaloids |2 NLM | |
| 700 | 1 | |a Bankoglu, Ezgi Eyluel |e verfasserin |4 aut | |
| 700 | 1 | |a Stopper, Helga |e verfasserin |4 aut | |
| 773 | 0 | 8 | |i Enthalten in |t Archives of toxicology |d 1974 |g 97(2023), 1 vom: 06. Jan., Seite 295-306 |w (DE-627)NLM000013374 |x 1432-0738 |7 nnns |
| 773 | 1 | 8 | |g volume:97 |g year:2023 |g number:1 |g day:06 |g month:01 |g pages:295-306 |
| 856 | 4 | 0 | |u http://dx.doi.org/10.1007/s00204-022-03394-z |3 Volltext |
| 912 | |a GBV_USEFLAG_A | ||
| 912 | |a GBV_NLM | ||
| 951 | |a AR | ||
| 952 | |d 97 |j 2023 |e 1 |b 06 |c 01 |h 295-306 | ||