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Establishment and Characterization of Three Afatinib-resistant Lung Adenocarcinoma PC-9 Cell Lines Developed with Increasing Doses of Afatinib
Published on: June 26, 2019
Biphasic function of GSK3β in gefitinib‑resistant NSCLC with or without EGFR mutations
Junzhe Li1, Xiayu Wu2, Xiang-Bo Ji3
1Department of Thoracic Surgery, Affiliated Cancer Hospital of Hainan Medical University, Haikou, Hainan 570312, P.R. China.
Abstract:
Epidermal growth factor receptor-tyrosine kinase inhibitors (EGFR-TKIs), such as gefitinib, are effective in the treatment of non-small cell lung cancer (NSCLC) harboring EGFR mutations. However, the mechanism underlying acquired resistance to EGFR-TKIs remains largely unknown. Therefore, the present study generated gefitinib-resistant PC-9 (PC-9G) cells, which were revealed to be more resistant to gefitinib-induced reductions in proliferation, migration and invasion, and increases in apoptosis, and had no detectable EGFR mutations compared with the control PC-9 cell line. In addition, the present study performed genome-wide transcriptomic analysis of differentially expressed genes between PC-9 and PC-9G cell lines. Cell proliferation, colony formation, invasion, migration and flow cytometry analyses were also performed. The genome-wide transcriptomic analysis revealed that glycogen synthase kinase 3β (GSK3β) was downregulated in PC-9G cells compared with that in PC-9 cells. Furthermore, GSK3β overexpression increased the proliferation, migration and invasion of PC-9 and H1975 gefitinib-resistant cells. Conversely, overexpression of GSK3β suppressed the proliferation, migration and invasion of PC-9G cells. Furthermore, AKT inhibition reduced the proliferation, migration and invasion, and induced the apoptosis of PC-9, PC-9G and H1975 cells, the effects of which were reversed following AKT activation; notably, the tumor suppressor function of GSK3β was inconsistent with the tumor promotor role of the AKT pathway in PC-9G cells without EGFR mutation. The present study may provide novel insights into the distinctive role of GSK3β in gefitinib-resistant NSCLC with or without EGFR mutations, suggesting that a more detailed investigation on GSK3β as a therapeutic target for gefitinib-resistant NSCLC may be warranted.
Insights
Acquired resistance to gefitinib in non-small cell lung cancer (NSCLC) involves changes in glycogen synthase kinase 3β (GSK3β). This study investigated GSK3β
Area of Science:
- Oncology
- Molecular Biology
- Pharmacology
Background:
- Epidermal growth factor receptor-tyrosine kinase inhibitors (EGFR-TKIs) like gefitinib are crucial for treating EGFR-mutated non-small cell lung cancer (NSCLC).
- Acquired resistance to EGFR-TKIs is a significant clinical challenge, and its underlying mechanisms, particularly in EGFR-wildtype NSCLC, are not fully understood.
- Understanding resistance mechanisms is vital for developing effective therapeutic strategies against NSCLC.
Purpose of the Study:
- To investigate the role of glycogen synthase kinase 3β (GSK3β) in acquired gefitinib resistance in non-small cell lung cancer (NSCLC).
- To compare gene expression profiles between gefitinib-sensitive and gefitinib-resistant NSCLC cell lines.
- To explore the functional impact of GSK3β and AKT pathway modulation on gefitinib-resistant NSCLC cells.
Main Methods:
- Generation of gefitinib-resistant PC-9 (PC-9G) cell line.
- Genome-wide transcriptomic analysis to identify differentially expressed genes.
- Functional assays including cell proliferation, colony formation, invasion, migration, and flow cytometry.
- Manipulation of GSK3β and AKT pathway activity through overexpression and inhibition/activation.
Main Results:
- Gefitinib-resistant PC-9G cells exhibited reduced sensitivity to gefitinib and showed no detectable EGFR mutations.
- Glycogen synthase kinase 3β (GSK3β) was significantly downregulated in gefitinib-resistant PC-9G cells compared to sensitive PC-9 cells.
- GSK3β overexpression promoted proliferation, migration, and invasion in gefitinib-resistant cells lacking EGFR mutations, while AKT pathway activation had a pro-tumor role, indicating a complex interplay.
Conclusions:
- Glycogen synthase kinase 3β (GSK3β) plays a distinct role in gefitinib-resistant non-small cell lung cancer (NSCLC), potentially acting as a tumor suppressor in EGFR-mutated contexts and a promoter in EGFR-wildtype resistant cells.
- The interplay between GSK3β and the AKT pathway is complex and context-dependent in gefitinib-resistant NSCLC.
- GSK3β warrants further investigation as a potential therapeutic target for overcoming gefitinib resistance in NSCLC.
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