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Erlotinib Activates Different Cell Death Pathways in EGFR-mutant Lung Cancer Cells Grown in 3D Versus 2D Culture
Hyun-Kyung Lee1, Min Hye Noh2, Seung-Woo Hong2
1Division of Pulmonary, Allergy, and Critical Care Medicine, Department of Internal Medicine, Inje University Busan Paik Hospital, Busan, Republic of Korea.
Background/Aim:
Non-small cell lung cancer patients with epidermal growth factor receptor (EGFR) mutation have been shown to have a good response to erlotinib, a receptor tyrosine kinase inhibitor of EGFR. In this study, we found that the cell death pathways activated by erlotinib in 2D and 3D culture systems are different.
Materials And Methods:
The cell death pathways induced by erlotinib were evaluated by flow cytometry and immunoblotting in both 2D and 3D culture systems of EGFR mutant lung cancer cells.
Results:
Treatment with erlotinib induced caspase 8 activation and up-regulation of TNF-related apoptosis-inducing ligand (TRAIL) expression only in 3D cultures. Knockdown of TRAIL attenuated both erlotinib-induced activation of caspase-8 and apoptosis in 3D cultures. Erlotinib also increased LC3, an autophagy marker, expression and c-Jun N terminal kinase (JNK) activation. Both 3-MA as an autophagy inhibitor and SP600125 as a JNK inhibitor, significantly inhibited erlotinib-induced cell death.
Conclusion:
Erlotinib induces apoptotic cell death in 3D cultures through an autophagy-TRAIL-JNK pathway.
Insights
Erlotinib triggers different cell death pathways in lung cancer cells depending on culture method. In 3D cultures, erlotinib induces apoptosis via autophagy, TRAIL, and JNK signaling.
Area of Science:
- Oncology
- Molecular Biology
- Cell Biology
Background:
- Epidermal growth factor receptor (EGFR) mutations are common in non-small cell lung cancer (NSCLC).
- Erlotinib, an EGFR inhibitor, shows efficacy in EGFR-mutated NSCLC.
- Cell death pathways activated by erlotinib may differ between 2D and 3D cell culture models.
Purpose of the Study:
- To investigate the distinct cell death pathways induced by erlotinib in 2D versus 3D culture systems of EGFR-mutated lung cancer cells.
- To elucidate the molecular mechanisms underlying erlotinib-induced cell death in a more physiologically relevant 3D environment.
Main Methods:
- Utilized flow cytometry and immunoblotting to assess cell death pathways.
- Employed EGFR-mutant lung cancer cell lines in both 2D and 3D culture models.
- Investigated the roles of caspase-8, TNF-related apoptosis-inducing ligand (TRAIL), autophagy markers (LC3), and c-Jun N-terminal kinase (JNK) signaling.
Main Results:
- Erlotinib induced caspase-8 activation and TRAIL expression specifically in 3D cultures.
- TRAIL knockdown reduced erlotinib-induced caspase-8 activation and apoptosis in 3D cultures.
- Erlotinib increased autophagy marker LC3 and JNK activation in 3D cultures.
- Autophagy inhibition (3-MA) and JNK inhibition (SP600125) significantly reduced erlotinib-induced cell death in 3D.
Conclusions:
- Erlotinib induces apoptotic cell death in 3D lung cancer cell cultures.
- This apoptosis is mediated by a novel pathway involving autophagy, TRAIL, and JNK signaling.
- Findings highlight the importance of 3D culture models for understanding drug-induced cell death mechanisms in NSCLC.
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