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Identification of AGR2 Gene-Specific Expression Patterns Associated with Epithelial-Mesenchymal Transition.

Andrea Martisova1,2, Lucia Sommerova1, Adam Krejci1

  • 1Research Centre for Applied Molecular Oncology (RECAMO), Masaryk Memorial Cancer Institute, Zluty Kopec 7, 65653 Brno, Czech Republic.

International Journal of Molecular Sciences
|September 23, 2022
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Summary

AGR2 downregulation and TGF-β signaling impact focal adhesion formation and cancer cell migration. AGR2 is also identified as a key player in arachidonic acid metabolism, affecting cancer progression.

Keywords:
AGR2EMTRNAseqTGF-βarachidonic acidfocal adhesion

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Area of Science:

  • Cell Biology
  • Molecular Oncology
  • Cancer Signaling Pathways

Background:

  • The TGF-β signaling pathway is crucial for cellular processes, and its dysregulation contributes to cancer development.
  • Epithelial to mesenchymal transition (EMT), a key process in tumor metastasis, is significantly influenced by TGF-β signaling.
  • AGR2 is recognized for maintaining epithelial phenotype and counteracting TGF-β-induced EMT.

Purpose of the Study:

  • To investigate the role of AGR2 in TGF-β-mediated cellular processes in lung cancer.
  • To identify molecular pathways affected by AGR2 knockout and TGF-β treatment in A549 lung cancer cells.
  • To elucidate the interplay between AGR2, TGF-β, focal adhesion, and arachidonic acid metabolism in cancer progression.

Main Methods:

  • Transcriptomic profiling of A549 lung cancer cells with CRISPR-Cas9 mediated AGR2 knockout, with and without TGF-β treatment.
  • Validation of key transcriptomic changes using RT-qPCR for focal adhesion and arachidonic acid metabolism markers.
  • Immunofluorescence microscopy to assess focal adhesion formation (stress fibers, vinculin foci).
  • Enzyme-linked immunosorbent assay (ELISA) to quantify prostaglandin E2 (PGE2) levels.

Main Results:

  • AGR2 knockout and TGF-β treatment significantly altered transcripts related to focal adhesion and arachidonic acid metabolism.
  • Validation confirmed changes in focal adhesion genes (e.g., COL4A1, FLNA, VINC) and stress fiber formation.
  • Downregulation of arachidonic acid metabolism transcripts (e.g., GPX2, PTGS2) and reduced PGE2 levels were observed.
  • Synergistic effects of AGR2 loss and TGF-β on focal adhesion formation and stress fiber development were noted.

Conclusions:

  • AGR2 downregulation and TGF-β signaling are critical for focal adhesion formation, influencing cancer cell migration and invasion.
  • AGR2 plays a significant role in the arachidonic acid metabolic pathway, impacting cancer progression.
  • The study highlights a novel connection between AGR2, TGF-β, focal adhesion dynamics, and eicosanoid metabolism in lung cancer.