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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
Establishment and Characterization of Three Afatinib-resistant Lung Adenocarcinoma PC-9 Cell Lines Developed with
Toshimitsu Yamaoka1, Motoi Ohba2, Yuki Matsunaga3
1Advanced Cancer Translational Research Institute, Showa University; yamaoka.t@med.showa-u.ac.jp.
Abstract:
Acquired resistance to molecular target inhibitors is a severe problem in cancer therapy. Lung cancer remains the leading cause of cancer-related death in most countries. The discovery of "oncogenic driver mutations," such as epidermal growth factor receptor (EGFR)-activating mutations, and subsequent development of molecular targeted agents of EGFR tyrosine kinase inhibitors (TKIs) (gefitinib, erlotinib, afatinib, dacomitinib, and osimertinib) have dramatically altered lung cancer treatment in recent decades. However, these drugs are still not effective in patients with non-small cell lung cancer (NSCLC) carrying EGFR-activating mutations. Following acquired resistance, the systemic progression of NSCLC remains a significant obstacle in treating patients with EGFR mutation-positive NSCLC. Here, we present a stepwise dose escalation method for establishing three independent acquired afatinib-resistant cell lines from NSCLC PC-9 cells harboring EGFR-activating mutations of 15-base pair deletions in EGFR exon 19. Methods for characterizing the three independent afatinib-resistance cell lines are briefly presented. The acquired resistance mechanisms to EGFR TKIs are heterogeneous. Therefore, multiple cell lines with acquired resistance to EGFR-TKIs must be examined. Ten to twelve months are required to obtain cell lines with acquired resistance using this stepwise dose escalation approach. The discovery of novel acquired resistance mechanisms will contribute to the development of more effective and safe therapeutic strategies.
Insights
Acquired resistance to EGFR TKIs is a major challenge in lung cancer. This study developed afatinib-resistant cell lines to investigate novel resistance mechanisms for better cancer therapies.
Area of Science:
- Oncology
- Molecular Biology
- Cancer Genetics
Background:
- Acquired resistance to molecular targeted inhibitors poses a significant challenge in cancer therapy, particularly for lung cancer.
- Epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIs) have revolutionized non-small cell lung cancer (NSCLC) treatment, but resistance remains an issue.
- EGFR-activating mutations drive NSCLC, and developing resistance to TKIs like afatinib necessitates understanding underlying mechanisms.
Purpose of the Study:
- To establish afatinib-resistant NSCLC cell lines using a stepwise dose escalation method.
- To investigate the heterogeneous mechanisms of acquired resistance to EGFR TKIs.
- To facilitate the discovery of novel resistance mechanisms for improved therapeutic strategies.
Main Methods:
- Established three independent afatinib-resistant NSCLC cell lines from PC-9 cells with EGFR exon 19 deletions via stepwise dose escalation.
- Characterized the acquired afatinib-resistant cell lines.
- The process required 10–12 months to generate resistant cell lines.
Main Results:
- Successfully generated three distinct afatinib-resistant NSCLC cell lines.
- Demonstrated the feasibility of a stepwise dose escalation approach for creating resistant models.
- Highlighted the heterogeneity of acquired resistance mechanisms to EGFR TKIs.
Conclusions:
- The development of afatinib-resistant cell lines is crucial for studying resistance mechanisms.
- Understanding acquired resistance heterogeneity is key to overcoming therapeutic limitations in EGFR-mutated NSCLC.
- This research contributes to developing more effective and safer treatment strategies for NSCLC patients.
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