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Rapid screening of drug candidates against EGFR/HER2 signaling pathway using fluorescence assay

Farkhondeh Khanjani1, Reza H Sajedi2, Sadegh Hasannia1

  • 1Department of Biochemistry, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, 14115-154, Iran.

Analytical and Bioanalytical Chemistry
|October 20, 2018
PubMed

Insights

This study introduces a new calcium-based assay for breast cancer drug discovery. The method effectively screens compounds targeting the human epidermal growth factor receptor 2 (HER2) pathway, showing promise for developing targeted therapies.

Area of Science:

  • Biochemistry
  • Cell Biology
  • Pharmacology

Background:

  • Calcium-based assays are increasingly popular for drug discovery.
  • Breast cancer necessitates rapid screening methods for effective drug candidates with minimal side effects.
  • Human epidermal growth factor receptor 2 (HER2) signaling involves intracellular calcium (Ca2+) release.

Purpose of the Study:

  • To develop and validate a simple, efficient, and specific calcium-based assay for screening drug candidates targeting the EGFR/HER2 signaling pathway.
  • To investigate the effect of lapatinib, a tyrosine kinase inhibitor, on Ca2+ release in HER2-overexpressing breast cancer cells.

Main Methods:

  • Utilized the BT474 cell line, which overexpresses the HER2 receptor.
  • Employed fura-2-AM to measure intracellular Ca2+ release by monitoring fluorescence intensity.
  • Administered epidermal growth factor (EGF) to stimulate the pathway and lapatinib to inhibit it.

Main Results:

  • Epidermal growth factor (EGF) stimulation led to a significant increase in intracellular Ca2+ fluorescence.
  • Treatment with lapatinib inhibited the EGFR/HER2 signaling pathway, decreasing Ca2+ influx and fluorescence.
  • The calculated IC50 value for lapatinib in BT474 cells was 113.2 nmol/L.

Conclusions:

  • The developed calcium-based assay is a simple, efficient, and specific method for evaluating drug candidates.
  • This assay is suitable for screening compounds that target the EGFR/HER2 heterodimer signaling pathway.
  • The findings support the potential utility of this assay in breast cancer drug discovery efforts.
Keywords:
Breast CancerEGFR/HER2 heterodimerFura-2-AMIntracellular Ca2+Tyrosine kinase inhibitor (TKI)

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