Related Experiment Video
Updated: Feb 20, 2026
Establishment and Characterization of Three Afatinib-resistant Lung Adenocarcinoma PC-9 Cell Lines Developed with Increasing Doses of Afatinib
Published on: June 26, 2019
Inhibitory Effect of Afatinib on Platelet Activation and Apoptosis
Hang Cao1, Abdulla Al Mamun Bhuyan1, Anja T Umbach1
1Department of Internal Medicine III, Tuebingen, Germany.
Background/Aims:
The epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor afatinib is used for the treatment of several malignancies. Afatinib is at least partially effective by triggering apoptosis of tumor cells. Platelets may similarly undergo apoptosis, which is characterized by caspase 3 activation, cell shrinkage and phosphatidylserine translocation. However, an effect of afatinib on platelets has never been reported. The present study explored whether treatment of platelets with afatinib modifies platelet activation and apoptosis in the absence and presence of platelet activators thrombin or collagen related peptide (CRP).
Methods:
Platelets isolated from wild-type mice were exposed for 30 minutes to afatinib (18 µg/ml) without or with subsequent treatment with thrombin (0.005 U/ml or 0.01 U/ml) or CRP (2 µg/ml or 5 µg/ml). Flow cytometry was employed to estimate Orai1 abundance at the platelet surface with specific antibodies, cytosolic Ca2+-activity ([Ca2+]i) from Fluo-3 fluorescence, platelet degranulation from P-selectin abundance, integrin activation from αIIbβ3 integrin abundance, caspase activity utilizing an Active Caspase-3 Staining kit, phosphatidylserine abundance from annexin-V-binding, platelet volume from forward scatter and aggregation utilizing staining with CD9-APC and CD9-PE.
Results:
In the absence of thrombin and CRP, the administration of afatinib (18 µg/ml) slightly, but significantly, increased [Ca2+]i and annexin-V-binding, but did not significantly modify Orai1 abundance, P-selectin abundance, activated αIIbβ3 integrin, cell volume, caspase activity and aggregation. Exposure of platelets to 0.005 U/ml or 0.01 U/ml thrombin or 2 µg/ml or 5 µg/ ml CRP was followed by a significant increase of Orai1 abundance, increase of [Ca2+]i, P-selectin abundance, αIIbβ3 integrin activity, annexin-V-binding, caspase activity, and aggregation, as well as a significant decrease of forward scatter, all effects significantly blunted (thrombin) or virtually abolished (CRP) by afatinib.
Conclusions:
Afatinib is a powerful inhibitor of platelet activation, platelet apoptosis and platelet aggregation.
Insights
Afatinib, an EGFR inhibitor, significantly inhibits platelet activation and apoptosis. This study reveals afatinib
Area of Science:
- Hematology
- Oncology
- Pharmacology
Background:
- Afatinib, an epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor, is used to treat various cancers.
- Afatinib's anti-cancer efficacy involves inducing tumor cell apoptosis.
- Platelet apoptosis shares characteristics with tumor cell apoptosis, but afatinib's effect on platelets is unstudied.
Purpose of the Study:
- To investigate the effects of afatinib on platelet activation and apoptosis.
- To determine if afatinib modulates platelet responses in the presence of activators like thrombin and collagen-related peptide (CRP).
Main Methods:
- Washed platelets from wild-type mice were treated with afatinib.
- Subsequent stimulation with thrombin or CRP was performed.
- Flow cytometry assessed platelet surface markers, intracellular calcium, degranulation, integrin activation, caspase activity, phosphatidylserine translocation, cell volume, and aggregation.
Main Results:
- Afatinib alone caused minor increases in intracellular calcium and phosphatidylserine binding.
- Afatinib significantly inhibited thrombin- and CRP-induced platelet activation markers, including degranulation, integrin activation, and aggregation.
- Afatinib markedly reduced thrombin- and CRP-induced platelet apoptosis markers, such as caspase activity and phosphatidylserine translocation.
Conclusions:
- Afatinib demonstrates potent inhibitory effects on platelet activation.
- Afatinib effectively suppresses platelet apoptosis.
- Afatinib significantly reduces platelet aggregation.
Related Concept Videos
01:20Antiplatelet Drugs: Prostaglandin Synthesis, P2Y12 and Glycoprotein IIb/IIIa Inhibitors
Prostaglandin synthesis inhibitors, exemplified by the widely known aspirin, wield their power by irreversibly acetylating...
01:17Intracellular Signaling Affects Focal Adhesions
Some...
01:22Formation of the Platelet Plug
As the injured blood vessel contracts, endothelial cells undergo contraction, revealing collagen fibers in the basement membrane and underlying connective tissue. Furthermore, the plasma membrane of endothelial cells becomes adhesive, preparing the site for platelet adhesion. Platelets...