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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
Remarkable inhibition effects of afatinib alone or combining with paclitaxel in esophageal squamous cell carcinoma
Li-Yan Yang1, Zhi-Jian Cheng2, Zou Liu1
1State Key Laboratory of Molecular Oncology, Center for Cancer Precision Medicine, National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China.
Background And Aim:
Chemotherapy drugs do not work well in esophageal squamous cell carcinoma (ESCC), and none of the targeted drugs have been applied in clinic. This study aims to identify effective targeted drugs and related biomarkers for the treatment of ESCC.
Methods:
The effect of 40 Food and Drug Administration-approved small-molecule inhibitors was first tested in five ESCC cell lines. CCK8 assays and xenografts derived from ESCC cell lines were performed to evaluate the anti-ESCC effects of inhibitors or chemotherapeutic agents in vitro and in vivo, respectively. Immunohistochemistry was utilized to analyze the p-EGFR expression in tissues. Western blot combining with gray analysis was conducted to detect the expression of interest protein. Flow cytometry and immunofluorescence assay were used to analyze apoptosis, cell cycle, and mitotic changes after drug treatment.
Results:
Afatinib showed remarkable effects on inhibiting ESCC cells with higher expression of p-EGFR. Results from combinatorial screening in ESCC cells expressing lower phosphorylation level of EGFR showed that paclitaxel and afatinib presented a significant synergistic inhibitory effect (P < 0.001). Molecular analysis revealed that paclitaxel sensitized afatinib by activating EGFR, and afatinib in combination with paclitaxel effectively blocked MAPK pathway and induced G2/M cell arrest and apoptosis that is an indicator of mitotic catastrophe.
Conclusions:
Our data demonstrate that afatinib is an effective drug for patients with ESCC expressing higher phosphorylation level of EGFR. And for patients with lower p-EGFR in tumors, paclitaxel in combination with afatinib might be a promising therapeutic strategy in ESCC.
Insights
Afatinib effectively treats esophageal squamous cell carcinoma (ESCC) with high p-EGFR. For ESCC with low p-EGFR, combining paclitaxel and afatinib shows promise.
Area of Science:
- Oncology
- Molecular Biology
- Pharmacology
Background:
- Chemotherapy is largely ineffective for esophageal squamous cell carcinoma (ESCC).
- Targeted therapies are not yet established for ESCC treatment.
- Identifying effective targeted drugs and biomarkers is crucial for ESCC management.
Purpose of the Study:
- To identify effective targeted drugs for esophageal squamous cell carcinoma (ESCC).
- To discover predictive biomarkers for targeted therapy response in ESCC.
- To evaluate novel therapeutic strategies for ESCC.
Main Methods:
- Screened 40 FDA-approved small-molecule inhibitors in five ESCC cell lines.
- Assessed anti-ESCC effects using CCK8 assays (in vitro) and xenografts (in vivo).
- Analyzed p-EGFR expression, protein levels, apoptosis, cell cycle, and mitotic changes via immunohistochemistry, Western blot, flow cytometry, and immunofluorescence.
Main Results:
- Afatinib demonstrated significant efficacy in ESCC cells with high p-EGFR expression.
- A combination of paclitaxel and afatinib showed synergistic inhibitory effects in ESCC cells with low EGFR phosphorylation.
- Paclitaxel sensitized afatinib by activating EGFR, leading to MAPK pathway inhibition, G2/M cell arrest, and apoptosis.
Conclusions:
- Afatinib is a potential treatment for ESCC patients with higher p-EGFR levels.
- Combination therapy with paclitaxel and afatinib may offer a promising strategy for ESCC patients with lower p-EGFR.
- EGFR phosphorylation status is a key biomarker for guiding ESCC treatment decisions.
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