Stimulation of Suicidal Erythrocyte Death by Ceritinib-Treatment of Human Erythrocytes
Background/Aims: The anaplastic lymphoma kinase (ALK) inhibitor ceritinib is utilized for the treatment of ALK positive non-small cell lung carcinoma. Side effects of the drug include decrease of blood hemoglobin concentration. Possible causes of anemia include stimulation of suicidal erythrocyte de...
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| Format: | Article |
| Language: | English |
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Cell Physiol Biochem Press GmbH & Co KG
2016-12-01
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| Series: | Cellular Physiology and Biochemistry |
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| Online Access: | http://www.karger.com/Article/FullText/453167 |
| _version_ | 1818324988662382592 |
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| author | Abdulla Al Mamun Bhuyan Elena Signoretto Rosi Bissinger Florian Lang |
| author_facet | Abdulla Al Mamun Bhuyan Elena Signoretto Rosi Bissinger Florian Lang |
| author_sort | Abdulla Al Mamun Bhuyan |
| collection | DOAJ |
| description | Background/Aims: The anaplastic lymphoma kinase (ALK) inhibitor ceritinib is utilized for the treatment of ALK positive non-small cell lung carcinoma. Side effects of the drug include decrease of blood hemoglobin concentration. Possible causes of anemia include stimulation of suicidal erythrocyte death or eryptosis, which is characterized by cell shrinkage and cell membrane scrambling with phosphatidylserine translocation to the erythrocyte surface. Signaling of eryptosis includes increase of cytosolic Ca2+ activity ([Ca2+]i), oxidative stress, ceramide, staurosporine sensitive protein kinase C, SB203580 sensitive p38 kinase, D4476 sensitive casein kinase 1, and zVAD sensitive caspases. The present study explored, whether ceritinib induces eryptosis and, if so, to shed light on the cellular mechanisms involved. Methods: Phosphatidylserine exposure at the cell surface was estimated from annexin-V-binding, cell volume from forward scatter, [Ca2+]i from Fluo3-fluorescence, ROS formation from DCFDA dependent fluorescence, and ceramide abundance utilizing specific antibodies. Results: A 48 hours exposure of human erythrocytes to ceritinib (1 µg/ml) significantly increased the percentage of annexin-V-binding cells, significantly decreased forward scatter, significantly increased Fluo3-fluorescence, but did not significantly modify DCFDA fluorescence or ceramide abundance. The effect of ceritinib on annexin-V-binding was significantly blunted but not abolished by removal of extracellular Ca2+, by the kinase inhibitors staurosporine (1 µM), SB203580 (2 µM) and D4476 (10 µM), as well as by caspase inhibitor zVAD (10 µM). Conclusions: Ceritinib triggers cell shrinkage and phospholipid scrambling of the erythrocyte cell membrane, an effect at least in part due to Ca2+ entry, as well as activation of kinases and Caspases. |
| first_indexed | 2024-12-13T11:37:20Z |
| format | Article |
| id | doaj.art-aaf119f915694e2fa8316ac1b7de3012 |
| institution | Directory Open Access Journal |
| issn | 1015-8987 1421-9778 |
| language | English |
| last_indexed | 2024-12-13T11:37:20Z |
| publishDate | 2016-12-01 |
| publisher | Cell Physiol Biochem Press GmbH & Co KG |
| record_format | Article |
| series | Cellular Physiology and Biochemistry |
| spelling | doaj.art-aaf119f915694e2fa8316ac1b7de30122022-12-21T23:47:45ZengCell Physiol Biochem Press GmbH & Co KGCellular Physiology and Biochemistry1015-89871421-97782016-12-014051129114010.1159/000453167453167Stimulation of Suicidal Erythrocyte Death by Ceritinib-Treatment of Human ErythrocytesAbdulla Al Mamun BhuyanElena SignorettoRosi BissingerFlorian LangBackground/Aims: The anaplastic lymphoma kinase (ALK) inhibitor ceritinib is utilized for the treatment of ALK positive non-small cell lung carcinoma. Side effects of the drug include decrease of blood hemoglobin concentration. Possible causes of anemia include stimulation of suicidal erythrocyte death or eryptosis, which is characterized by cell shrinkage and cell membrane scrambling with phosphatidylserine translocation to the erythrocyte surface. Signaling of eryptosis includes increase of cytosolic Ca2+ activity ([Ca2+]i), oxidative stress, ceramide, staurosporine sensitive protein kinase C, SB203580 sensitive p38 kinase, D4476 sensitive casein kinase 1, and zVAD sensitive caspases. The present study explored, whether ceritinib induces eryptosis and, if so, to shed light on the cellular mechanisms involved. Methods: Phosphatidylserine exposure at the cell surface was estimated from annexin-V-binding, cell volume from forward scatter, [Ca2+]i from Fluo3-fluorescence, ROS formation from DCFDA dependent fluorescence, and ceramide abundance utilizing specific antibodies. Results: A 48 hours exposure of human erythrocytes to ceritinib (1 µg/ml) significantly increased the percentage of annexin-V-binding cells, significantly decreased forward scatter, significantly increased Fluo3-fluorescence, but did not significantly modify DCFDA fluorescence or ceramide abundance. The effect of ceritinib on annexin-V-binding was significantly blunted but not abolished by removal of extracellular Ca2+, by the kinase inhibitors staurosporine (1 µM), SB203580 (2 µM) and D4476 (10 µM), as well as by caspase inhibitor zVAD (10 µM). Conclusions: Ceritinib triggers cell shrinkage and phospholipid scrambling of the erythrocyte cell membrane, an effect at least in part due to Ca2+ entry, as well as activation of kinases and Caspases.http://www.karger.com/Article/FullText/453167PhosphatidylserineCell volumeEryptosisIonomycinCalcium |
| spellingShingle | Abdulla Al Mamun Bhuyan Elena Signoretto Rosi Bissinger Florian Lang Stimulation of Suicidal Erythrocyte Death by Ceritinib-Treatment of Human Erythrocytes Cellular Physiology and Biochemistry Phosphatidylserine Cell volume Eryptosis Ionomycin Calcium |
| title | Stimulation of Suicidal Erythrocyte Death by Ceritinib-Treatment of Human Erythrocytes |
| title_full | Stimulation of Suicidal Erythrocyte Death by Ceritinib-Treatment of Human Erythrocytes |
| title_fullStr | Stimulation of Suicidal Erythrocyte Death by Ceritinib-Treatment of Human Erythrocytes |
| title_full_unstemmed | Stimulation of Suicidal Erythrocyte Death by Ceritinib-Treatment of Human Erythrocytes |
| title_short | Stimulation of Suicidal Erythrocyte Death by Ceritinib-Treatment of Human Erythrocytes |
| title_sort | stimulation of suicidal erythrocyte death by ceritinib treatment of human erythrocytes |
| topic | Phosphatidylserine Cell volume Eryptosis Ionomycin Calcium |
| url | http://www.karger.com/Article/FullText/453167 |
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