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Updated: Apr 26, 2026
Establishing Dual Resistance to EGFR-TKI and MET-TKI in Lung Adenocarcinoma Cells In Vitro with a 2-step Dose-escalation Procedure
Published on: August 11, 2017
In vitro study on blocking mTOR signaling pathway in EGFR-TKI resistance NSCLC
Xu-Dong Xiang1, Jing Yu2, Gao-Feng Li1
1Department of Thoracic Surgery, Third Affiliated Hospital of Kunming Medical University, Kunming 650118, China.
Objective:
To investigate the effect and mechanism of inhibitor everolimus on EGFR-TKI resistance NSCLC.
Methods:
MTT assay was used to detect proliferation of human non-small cell lung cancer cell line A549. Flow cytometry was used to detect the changes of apoptosis and cycle distribution in each group after 24 h and 48 h. RT-PCR was used to detect the changes of PTEN and 4EBP1 expression levels after 48 h of monotherapy and combination therapy.
Results:
MTT assay showed that everolimus had dose-dependent inhibition against growth of A549 cells. Flow cytometry showed when everolimus could induce apoptosis and induce G0/G1 phase cell cycle arrest, which was time-dependent (P<0.05). RT-PCR showed everolimus could increase PTEN and 4EBP1 expression.
Conclusions:
mTOR inhibitor everolimus has an inhibitory effect on EGFR-TKI resistant NSCLC, which cannot reverse the resistance effect of EGFR-TKI resistant cell line A549. The relationship between EGFR/AKT signaling pathway and the mTOR signaling pathway and the mechanism in non-small cell lung cancer need further study.
Insights
The mTOR inhibitor everolimus shows inhibitory effects on EGFR-TKI resistant non-small cell lung cancer (NSCLC) cells. However, it does not reverse resistance in the A549 cell line, indicating further research is needed.
Area of Science:
- Oncology
- Molecular Biology
- Pharmacology
Background:
- Non-small cell lung cancer (NSCLC) often develops resistance to Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitors (EGFR-TKIs).
- Understanding resistance mechanisms is crucial for developing effective treatments for NSCLC.
Purpose of the Study:
- To investigate the effect of the mTOR inhibitor everolimus on EGFR-TKI resistant NSCLC.
- To elucidate the underlying mechanisms of everolimus's action in resistant NSCLC.
Main Methods:
- MTT assay was used to assess the proliferation of the A549 NSCLC cell line.
- Flow cytometry was employed to analyze apoptosis and cell cycle distribution.
- RT-PCR was utilized to measure the expression levels of PTEN and 4EBP1.
Main Results:
- Everolimus demonstrated a dose-dependent inhibition of A549 cell growth.
- Everolimus induced apoptosis and G0/G1 phase cell cycle arrest in a time-dependent manner.
- Everolimus treatment led to increased expression of PTEN and 4EBP1.
Conclusions:
- Everolimus, an mTOR inhibitor, exhibits an inhibitory effect on EGFR-TKI resistant NSCLC.
- Everolimus did not reverse the resistance in the EGFR-TKI resistant A549 cell line.
- Further investigation is required to understand the interplay between EGFR/AKT and mTOR signaling pathways in NSCLC resistance.
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