Related Experiment Video

Updated: May 24, 2025

Evaluation of Keratinocyte Proliferation on Two- and Three-dimensional Type I Collagen Substrates
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Evaluation of Keratinocyte Proliferation on Two- and Three-dimensional Type I Collagen Substrates

Published on: April 22, 2019

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Brigatinib can inhibit proliferation and induce apoptosis of human immortalized keratinocyte cells

Qi Yang1, Dan Zhao1, Linjie Ju2

  • 1School of Pharmacy, Nanjing University of Chinese Medicine, Nanjing, China.

Frontiers in Pharmacology
|March 5, 2025
PubMed
Abstract

Insights

Brigatinib, used for ALK-positive NSCLC, causes skin toxicities. This study shows brigatinib inhibits keratinocyte growth and apoptosis, potentially via the PI3K/AKT pathway, offering insights into managing these side effects.

Area of Science:

  • Dermatology
  • Oncology
  • Molecular Biology

Background:

  • Brigatinib is an effective treatment for anaplastic lymphoma kinase (ALK)-positive non-small cell lung cancer (NSCLC).
  • Dermal toxicities are a significant side effect of brigatinib treatment, but the underlying mechanisms are not fully understood.
  • Investigating the cellular effects of brigatinib on skin cells is crucial for managing treatment-related adverse events.

Purpose of the Study:

  • To elucidate the mechanism of brigatinib-induced dermal toxicities at the cellular level.
  • To evaluate the impact of brigatinib on human immortalized keratinocyte (HaCaT) cell proliferation, apoptosis, and cell cycle.
  • To identify key molecular pathways and genes involved in brigatinib's effects on keratinocytes.

Main Methods:

  • Cell proliferation was assessed using CCK-8, colony formation, and EdU incorporation assays.
  • Apoptosis was measured by Annexin FITC/PI and Acridine Orange staining.
  • Transcriptome sequencing (RNA-seq) was employed to identify differentially expressed genes and signaling pathways, followed by qRT-PCR and western blot validation of key targets including EGFR ligands and the PI3K/AKT pathway.

Main Results:

  • Brigatinib demonstrated significant inhibition of HaCaT cell proliferation with an IC50 of 2.9 μmol/L and increased apoptosis rates.
  • RNA-seq analysis revealed that brigatinib downregulates the expression of amphiregulin, epiregulin, and TGFA.
  • Protein analysis confirmed reduced levels of amphiregulin, epiregulin, TGFA, PI3K, AKT, and phosphorylated AKT (p-AKT) following brigatinib treatment.

Conclusions:

  • Brigatinib inhibits the growth and progression of human keratinocytes (HaCaT cells).
  • The study identifies downregulation of EGFR ligands (amphiregulin, epiregulin, TGFA) and the PI3K/AKT signaling pathway as key effects of brigatinib.
  • The PI3K/AKT pathway represents a potential therapeutic target for mitigating brigatinib-induced dermal toxicities.
Keywords:
PI3K/AKTTGFAamphiregulinbrigatinibepiregulin

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