Molecular profiling of afatinib-resistant non-small cell lung cancer cells in vivo derived from mice
Cheng-Ta Chung1, Kai-Chia Yeh2, Chia-Huei Lee3
1Graduate Institute of Biotechnology, National Tsing Hua University, Hsinchu, Taiwan; Institute of Biotechnology and Pharmaceutical Research, National Health Research Institutes, Zhunan, Taiwan.
Abstract:
Non-small-cell lung cancer (NSCLC) is a leading cause of cancer-related death worldwide. NSCLC patients with overexpressed or mutated epidermal growth factor receptor (EGFR) related to disease progression are treated with EGFR-tyrosine kinase inhibitors (EGFR-TKIs). Acquired drug resistance after TKI treatments has been a major focus for development of NSCLC therapies. This study aimed to establish afatinib-resistant cell lines from which afatinib resistance-associated genes are identified and the underlying mechanisms of multiple-TKI resistance in NSCLC can be further investigated. Nude mice bearing subcutaneous NSCLC HCC827 tumors were administered with afatinib at different dose intensities (5-100 mg/kg). We established three HCC827 sublines resistant to afatinib (IC50 > 1 μM) with cross-resistance to gefitinib (IC50 > 5 μM). cDNA microarray revealed several of these sublines shared 27 up- and 13 down-regulated genes. The mRNA expression of selective novel genes - such as transmembrane 4 L six family member 19 (TM4SF19), suppressor of cytokine signaling 2 (SOCS2), and quinolinate phosphoribosyltransferase (QPRT) - are responsive to afatinib treatments only at high concentrations. Furthermore, c-MET amplification and activations of a subset of tyrosine kinase receptors were observed in all three resistant cells. PHA665752, a c-MET inhibitor, remarkably increased the sensitivity of these resistant cells to afatinib (IC50 = 12-123 nM). We established afatinib-resistant lung cancer cell lines and here report genes associated with afatinib resistance in human NSCLC. These cell lines and the identified genes serve as useful investigational tools, prognostic biomarkers of TKI therapies, and promising molecule targets for development of human NSCLC therapeutics.
Insights
Researchers developed afatinib-resistant non-small-cell lung cancer (NSCLC) cell lines to identify resistance genes. These findings offer new targets for overcoming TKI resistance in NSCLC treatment.
Area of Science:
- Oncology
- Molecular Biology
- Pharmacology
Background:
- Non-small-cell lung cancer (NSCLC) is a major global cancer burden.
- Epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIs) are standard NSCLC treatments.
- Acquired TKI resistance limits therapeutic efficacy, necessitating research into resistance mechanisms.
Purpose of the Study:
- To establish afatinib-resistant NSCLC cell lines.
- To identify genes associated with afatinib resistance.
- To investigate mechanisms of multiple-TKI resistance in NSCLC.
Main Methods:
- Generation of afatinib-resistant HCC827 NSCLC sublines in nude mice.
- cDNA microarray analysis to identify differentially expressed genes.
- Assessment of c-MET amplification and tyrosine kinase receptor activation.
- Evaluation of c-MET inhibitor PHA665752 in resistant cells.
Main Results:
- Established three afatinib-resistant HCC827 sublines with cross-resistance to gefitinib.
- Identified 27 up-regulated and 13 down-regulated genes in resistant sublines.
- Observed c-MET amplification and activated tyrosine kinase receptors in resistant cells.
- PHA665752 significantly resensitized resistant cells to afatinib.
Conclusions:
- Afatinib-resistant NSCLC cell lines were successfully established.
- Novel genes (e.g., TM4SF19, SOCS2, QPRT) are associated with afatinib resistance.
- c-MET pathway activation contributes to TKI resistance.
- Identified genes and cell lines are valuable tools for NSCLC research and therapeutic development.
