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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
Preclinical evaluation of afatinib (BIBW2992) in esophageal squamous cell carcinoma (ESCC)
Chi Hang Wong1, Brigette Buig Yue Ma1, Connie Wun Chun Hui1
1State Key Laboratory of Oncology in South China, Sir Y.K. Pao Centre for Cancer, Department of Clinical Oncology, Cancer Drug Testing Unit, Hong Kong Cancer Institute and Li Ka Shing Institute of Health Sciences, The Chinese University of Hong Kong Shatin, New Territories, Hong Kong SAR, China.
Abstract:
Esophageal squamous cell carcinoma (ESCC) is the eighth most common cancer worldwide. Epidermal growth factor receptors (EGFR) are often overexpressed in esophageal cancers, thus anti-EGFR inhibitors have been evaluated in ESCC. Afatinib was an irreversible inhibitor of these ErbB family receptors. This study characterized the preclinical activity of afatinib in five ESCC cell lines: HKESC-1, HKESC-2, KYSE510, SLMT-1 and EC-1. ESCC cell lines were sensitive to afatinib with IC50 concentrations at lower micro-molar range (at 72 hour incubation: HKESC-1 = 0.002 μM, HKESC-2 = 0.002 μM, KYSE510 = 1.090 μM, SLMT-1 = 1.161 μM and EC-1 = 0.109 μM) with a maximum growth inhibition over 95%. Afatinib can strongly induce G0/G1 cell cycle arrest in HKESC-2 and EC-1 in a dose- and time-dependent manner. The phosphorylation of ErbB family downstream effectors such as pAKT, pS6 and pMAPK were significantly inhibited in HKESC-2 and EC-1. Apoptosis was observed in both cell lines at 24 hours after exposure to afatinib, as determined by the presence of cleaved PARP. Afatinib could effectively inhibit HKESC-2 tumor growth in mice without obvious toxicity. Afatinib alone has shown excellent growth inhibitory effect on ESCC in both in vitro and in vivo models, however, no synergistic effect was observed when it was combined with chemotherapeutic agents such as 5-fluorouracil (5-FU) and cisplatin. In summary, afatinib can inhibit cell proliferation effectively by arresting the cells in G0/G1 phase, as well as inducing apoptosis in ESCC. These findings warrant further studies of afatinib as therapeutic agent in treating ESCC.
Insights
Afatinib effectively inhibits esophageal squamous cell carcinoma (ESCC) growth by halting cell cycles and inducing apoptosis. This targeted therapy shows promise in preclinical models, warranting further investigation for ESCC treatment.
Area of Science:
- Oncology
- Molecular Biology
- Pharmacology
Background:
- Esophageal squamous cell carcinoma (ESCC) is a prevalent cancer globally.
- Epidermal growth factor receptor (EGFR) overexpression is common in ESCC, making anti-EGFR therapies a focus.
- Afatinib, an irreversible ErbB family inhibitor, has potential therapeutic applications.
Purpose of the Study:
- To evaluate the preclinical efficacy of afatinib in multiple ESCC cell lines.
- To investigate the mechanisms underlying afatinib's anti-cancer effects.
- To assess afatinib's therapeutic potential in an in vivo ESCC model.
Main Methods:
- Testing afatinib's inhibitory concentration (IC50) and growth inhibition across five ESCC cell lines.
- Analyzing afatinib's impact on cell cycle progression (G0/G1 arrest) and downstream signaling pathways (pAKT, pS6, pMAPK).
- Assessing afatinib-induced apoptosis via cleaved PARP and evaluating in vivo efficacy and toxicity in a mouse model.
Main Results:
- Afatinib demonstrated sensitivity across ESCC cell lines, with IC50 values in the lower micromolar range and over 95% growth inhibition.
- Afatinib induced significant G0/G1 cell cycle arrest and inhibited key downstream signaling pathways in sensitive cell lines.
- Apoptosis was confirmed, and afatinib effectively suppressed tumor growth in mice with minimal toxicity.
- No synergistic effects were observed when afatinib was combined with 5-fluorouracil or cisplatin.
Conclusions:
- Afatinib exhibits potent preclinical anti-cancer activity against ESCC through cell cycle arrest and apoptosis induction.
- Afatinib demonstrates efficacy in both in vitro and in vivo ESCC models, suggesting its potential as a monotherapy.
- Further clinical studies are warranted to explore afatinib's therapeutic role in treating esophageal squamous cell carcinoma.
