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CRISPR/Cas9 engineering of ERK5 identifies its FAK/PYK2 dependent role in adhesion-mediated cell survival

Moazzam Ali1, Zeeshan Mutahir1, Anjum Riaz1

  • 1Institute of Biochemistry and Biotechnology, University of the Punjab, 54590, Lahore, Pakistan.

Biochemical and Biophysical Research Communications
|April 7, 2019
PubMed

Insights

Extracellular signal-regulated kinase 5 (ERK5) is crucial for breast cancer cell survival and adhesion. Its absence, particularly when focal adhesion kinase (FAK) and proline-rich tyrosine kinase 2 (PYK2) are inhibited, significantly reduces cell viability and increases apoptosis.

Area of Science:

  • Cell Biology
  • Molecular Oncology
  • Cancer Signaling Pathways

Background:

  • Extracellular signal-regulated kinase 5 (ERK5) is recognized for its role in breast cancer cell proliferation, migration, and invasion.
  • ERK5 is also involved in anti-apoptotic signaling pathways induced by growth factors.
  • The specific contribution of ERK5 to adhesion-induced survival signaling remains unclear.

Purpose of the Study:

  • To investigate the role of ERK5 in adhesion-mediated cell viability in breast cancer.
  • To elucidate the signaling pathways involving ERK5, focal adhesion kinase (FAK), and proline-rich tyrosine kinase 2 (PYK2) in cell survival.

Main Methods:

  • CRISPR/Cas9 gene editing was employed to knock out ERK5 expression in cancer cell lines.
  • MDA-MB 231 breast cancer cells lacking ERK5 were utilized to assess adhesion-mediated cell viability.
  • Chemical inhibition of FAK and PYK2 using PF 431396 was performed.

Main Results:

  • ERK5-deficient cells showed reduced attachment to extracellular matrix proteins fibronectin and vitronectin.
  • Combined ERK5 knockout and FAK/PYK2 inhibition led to further decreased cell adhesion.
  • These cells exhibited significantly reduced cell viability and increased apoptosis, indicated by cleaved PARP and caspase 9 detection.

Conclusions:

  • ERK5 plays a pro-survival role in response to cell adhesion in breast cancer cells.
  • This pro-survival function of ERK5 is regulated by focal adhesion kinase (FAK) and proline-rich tyrosine kinase 2 (PYK2).
  • Targeting ERK5 in combination with FAK/PYK2 inhibition may represent a therapeutic strategy for breast cancer.
Keywords:
Breast cancerCRISPR/Cas9Cell adhesionCell survivalExtracellular signal-regulated kinase 5

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