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Analytical and Bioanalytical Chemistry

, Volume 410, Issue 30, pp 7827–7835 | Cite as

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

  • Farkhondeh Khanjani
  • Reza H. SajediEmail author
  • Sadegh Hasannia
Paper in Forefront

Abstract

Over the recent decade, the calcium-based assays have gained much popularity in order to discover new drugs. Since breast cancer is the second cause of death in the female population, rapid and effective methods are needed to screen drug compounds with fewer side effects. Human epidermal growth factor receptor 2 (HER2) increases intracellular free Ca2+ on its signaling pathways. In the present study, BT474 cell line, which overexpresses HER2 receptor, was selected and using fura-2-AM, intracellular Ca2+ release was investigated. The changes in the concentration of intracellular Ca2+ were evaluated by variation in the amount of fluorescence intensity. In the presence of epidermal growth factor (EGF), an increase in fluorescence intensity was observed so that after 20 min it raised to the maximum level. After treatment of BT474 cells by lapatinib, as a tyrosine kinase inhibitor (TKI), the signaling pathway of EGFR/HER2 heterodimer was significantly inhibited, which resulted in a decrease in Ca2+ entry into the cytoplasm and fluorescence emission decreased. The IC50 value for the effect of lapatinib on BT474 cells was 113.2 nmol/L. Our results suggest this method is a simple, efficient and specific approach and can potentially be useful for screening new drug candidates against EGFR/HER2 heterodimer signaling pathways.

Graphical abstract

Keywords

Breast Cancer EGFR/HER2 heterodimer Fura-2-AM Intracellular Ca2+ Tyrosine kinase inhibitor (TKI) 

Notes

Acknowledgments

This work was supported by the research council of Tarbiat Modares University and Ministry of Sciences, Researches, and Technology, Iran.

Compliance with ethical standards

Conflict of Interest

The authors declare that they have no conflict of interest.

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Farkhondeh Khanjani
    • 1
  • Reza H. Sajedi
    • 1
    Email author
  • Sadegh Hasannia
    • 1
  1. 1.Department of Biochemistry, Faculty of Biological SciencesTarbiat Modares UniversityTehranIran

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