Lapatinib Activates the Kelch-Like ECH-Associated Protein 1-Nuclear Factor Erythroid 2-Related Factor 2 Pathway in HepG2 Cells
The receptor tyrosine kinase inhibitor lapatinib, indicated to treat patients with HER2-positive breast cancer in combination with capecitabine, can cause severe hepatotoxicity. Lapatinib is further associated with mitochondrial toxicity and accumulation of reactive oxygen species. The effect of lap...
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doaj-304488ceebe54538b7c33266ec95e3f02020-11-25T03:50:17ZengFrontiers Media S.A.Frontiers in Pharmacology1663-98122020-06-011110.3389/fphar.2020.00944544552Lapatinib Activates the Kelch-Like ECH-Associated Protein 1-Nuclear Factor Erythroid 2-Related Factor 2 Pathway in HepG2 CellsNoëmi Johanna Roos0Noëmi Johanna Roos1Noëmi Johanna Roos2Diell Aliu3Diell Aliu4Jamal Bouitbir5Jamal Bouitbir6Jamal Bouitbir7Stephan Krähenbühl8Stephan Krähenbühl9Stephan Krähenbühl10Division of Clinical Pharmacology & Toxicology, University Hospital, Basel, SwitzerlandDepartment of Biomedicine, University of Basel, Basel, SwitzerlandSwiss Centre for Applied Human Toxicology (SCAHT), Basel, SwitzerlandDivision of Clinical Pharmacology & Toxicology, University Hospital, Basel, SwitzerlandDepartment of Biomedicine, University of Basel, Basel, SwitzerlandDivision of Clinical Pharmacology & Toxicology, University Hospital, Basel, SwitzerlandDepartment of Biomedicine, University of Basel, Basel, SwitzerlandSwiss Centre for Applied Human Toxicology (SCAHT), Basel, SwitzerlandDivision of Clinical Pharmacology & Toxicology, University Hospital, Basel, SwitzerlandDepartment of Biomedicine, University of Basel, Basel, SwitzerlandSwiss Centre for Applied Human Toxicology (SCAHT), Basel, SwitzerlandThe receptor tyrosine kinase inhibitor lapatinib, indicated to treat patients with HER2-positive breast cancer in combination with capecitabine, can cause severe hepatotoxicity. Lapatinib is further associated with mitochondrial toxicity and accumulation of reactive oxygen species. The effect of lapatinib on the Kelch-like ECH-associated protein 1 (Keap1)-nuclear factor erythroid 2-related factor 2 (Nrf2) pathway, the major cellular defense pathway against oxidative stress, has so far not been studied in detail. In the present study, we show that lapatinib (2–20 µM) activates the Keap1-Nrf2 pathway in HepG2 cells, a hepatocellular carcinoma-derived cell line, in a concentration-dependent manner upon 24 h of treatment. Lapatinib stabilized the transcription factor Nrf2 at concentrations ≥5 µM and caused its nuclear translocation. Well-established Nrf2 regulated genes (Nqo1, Gsta1, Gclc, and Gclm) were upregulated at lapatinib concentrations ≥10 µM. Furthermore, cellular and mitochondrial glutathione (GSH) levels increased starting at 10 µM lapatinib. As a marker of oxidative stress, cellular GSSG significantly increased at 10 and 20 µM lapatinib. Furthermore, the gene expression of mitochondrial Glrx2 and SOD2 were increased upon lapatinib treatment, which was also observed for the mitochondrial SOD2 protein content. In conclusion, lapatinib treatment for 24 h activated the Keap1-Nrf2 pathway in HepG2 cells starting at 10 μM, which is a clinically relevant concentration. As a consequence, treatment with lapatinib increased the mRNA and protein expression of antioxidative and other cytoprotective genes and induced GSH synthesis, but these measures could not completely block the oxidative stress associated with lapatinib.https://www.frontiersin.org/article/10.3389/fphar.2020.00944/fulllapatinibHepG2 cellsmitochondriareactive oxygen speciesNrf2Keap1 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Noëmi Johanna Roos Noëmi Johanna Roos Noëmi Johanna Roos Diell Aliu Diell Aliu Jamal Bouitbir Jamal Bouitbir Jamal Bouitbir Stephan Krähenbühl Stephan Krähenbühl Stephan Krähenbühl |
spellingShingle |
Noëmi Johanna Roos Noëmi Johanna Roos Noëmi Johanna Roos Diell Aliu Diell Aliu Jamal Bouitbir Jamal Bouitbir Jamal Bouitbir Stephan Krähenbühl Stephan Krähenbühl Stephan Krähenbühl Lapatinib Activates the Kelch-Like ECH-Associated Protein 1-Nuclear Factor Erythroid 2-Related Factor 2 Pathway in HepG2 Cells Frontiers in Pharmacology lapatinib HepG2 cells mitochondria reactive oxygen species Nrf2 Keap1 |
author_facet |
Noëmi Johanna Roos Noëmi Johanna Roos Noëmi Johanna Roos Diell Aliu Diell Aliu Jamal Bouitbir Jamal Bouitbir Jamal Bouitbir Stephan Krähenbühl Stephan Krähenbühl Stephan Krähenbühl |
author_sort |
Noëmi Johanna Roos |
title |
Lapatinib Activates the Kelch-Like ECH-Associated Protein 1-Nuclear Factor Erythroid 2-Related Factor 2 Pathway in HepG2 Cells |
title_short |
Lapatinib Activates the Kelch-Like ECH-Associated Protein 1-Nuclear Factor Erythroid 2-Related Factor 2 Pathway in HepG2 Cells |
title_full |
Lapatinib Activates the Kelch-Like ECH-Associated Protein 1-Nuclear Factor Erythroid 2-Related Factor 2 Pathway in HepG2 Cells |
title_fullStr |
Lapatinib Activates the Kelch-Like ECH-Associated Protein 1-Nuclear Factor Erythroid 2-Related Factor 2 Pathway in HepG2 Cells |
title_full_unstemmed |
Lapatinib Activates the Kelch-Like ECH-Associated Protein 1-Nuclear Factor Erythroid 2-Related Factor 2 Pathway in HepG2 Cells |
title_sort |
lapatinib activates the kelch-like ech-associated protein 1-nuclear factor erythroid 2-related factor 2 pathway in hepg2 cells |
publisher |
Frontiers Media S.A. |
series |
Frontiers in Pharmacology |
issn |
1663-9812 |
publishDate |
2020-06-01 |
description |
The receptor tyrosine kinase inhibitor lapatinib, indicated to treat patients with HER2-positive breast cancer in combination with capecitabine, can cause severe hepatotoxicity. Lapatinib is further associated with mitochondrial toxicity and accumulation of reactive oxygen species. The effect of lapatinib on the Kelch-like ECH-associated protein 1 (Keap1)-nuclear factor erythroid 2-related factor 2 (Nrf2) pathway, the major cellular defense pathway against oxidative stress, has so far not been studied in detail. In the present study, we show that lapatinib (2–20 µM) activates the Keap1-Nrf2 pathway in HepG2 cells, a hepatocellular carcinoma-derived cell line, in a concentration-dependent manner upon 24 h of treatment. Lapatinib stabilized the transcription factor Nrf2 at concentrations ≥5 µM and caused its nuclear translocation. Well-established Nrf2 regulated genes (Nqo1, Gsta1, Gclc, and Gclm) were upregulated at lapatinib concentrations ≥10 µM. Furthermore, cellular and mitochondrial glutathione (GSH) levels increased starting at 10 µM lapatinib. As a marker of oxidative stress, cellular GSSG significantly increased at 10 and 20 µM lapatinib. Furthermore, the gene expression of mitochondrial Glrx2 and SOD2 were increased upon lapatinib treatment, which was also observed for the mitochondrial SOD2 protein content. In conclusion, lapatinib treatment for 24 h activated the Keap1-Nrf2 pathway in HepG2 cells starting at 10 μM, which is a clinically relevant concentration. As a consequence, treatment with lapatinib increased the mRNA and protein expression of antioxidative and other cytoprotective genes and induced GSH synthesis, but these measures could not completely block the oxidative stress associated with lapatinib. |
topic |
lapatinib HepG2 cells mitochondria reactive oxygen species Nrf2 Keap1 |
url |
https://www.frontiersin.org/article/10.3389/fphar.2020.00944/full |
work_keys_str_mv |
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