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Updated: Apr 21, 2026

Establishment and Characterization of Three Afatinib-resistant Lung Adenocarcinoma PC-9 Cell Lines Developed with Increasing Doses of Afatinib
09:38

Establishment and Characterization of Three Afatinib-resistant Lung Adenocarcinoma PC-9 Cell Lines Developed with Increasing Doses of Afatinib

Published on: June 26, 2019

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Afatinib resistance in non-small cell lung cancer involves the PI3K/AKT and MAPK/ERK signalling pathways and

Simona Coco1, Anna Truini, Angela Alama

  • 1Lung Cancer Unit, IRCCS AOU San Martino-IST, L.go R. Benzi 10, 16132, Genoa, Italy, simona.coco@hsanmartino.it.

Targeted Oncology
|October 25, 2014
PubMed

Insights

Afatinib resistance in non-small cell lung cancer (NSCLC) may stem from activated epidermal growth factor receptor (EGFR) downstream pathways, not just mutations. Epithelial-to-mesenchymal transition also contributes to afatinib resistance.

Area of Science:

  • Oncology
  • Molecular Biology
  • Genetics

Background:

  • Epidermal growth factor receptor (EGFR) signaling is frequently dysregulated in non-small cell lung cancer (NSCLC).
  • Irreversible tyrosine kinase inhibitors (TKIs) like afatinib improve survival in advanced NSCLC with EGFR mutations.
  • Acquired resistance to TKIs remains a significant clinical challenge, limiting treatment efficacy.

Purpose of the Study:

  • To investigate afatinib sensitivity in EGFR-mutated NSCLC cell lines.
  • To analyze gene expression profiles and identify mechanisms of afatinib resistance.
  • To explore the role of EGFR downstream pathways and epithelial-to-mesenchymal transition in resistance.

Main Methods:

  • Expression profile analysis of 92 EGF pathway genes in NCI-H1650 and NCI-H1975 NSCLC cell lines.
  • Genomic analysis using array comparative genomic hybridization (aCGH) and deep sequencing.
  • Transcriptomic analysis via quantitative polymerase chain reaction (qPCR).
  • Proteomic analysis using Western blot and immunofluorescence.

Main Results:

  • NCI-H1650 cells, less responsive to afatinib, showed baseline activation of PI3K/AKT and PLCγ/PKC pathways.
  • Afatinib-resistant cells exhibited activation of PI3K/AKT and MAPK/ERK pathways.
  • A switch to an epithelial-to-mesenchymal phenotype was observed in resistant cells, potentially conferring resistance properties.

Conclusions:

  • Activation of EGFR-dependent downstream pathways, including PI3K/AKT and MAPK/ERK, is implicated in afatinib resistance.
  • Epithelial-to-mesenchymal transition may represent a novel mechanism underlying afatinib resistance in NSCLC.
  • EGFR mutational status alone may be insufficient for selecting TKI treatments; downstream pathway activation should also be considered.

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