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Loss of transcription factor EB dysregulates the G1/S transition and DNA replication in mammary epithelial cells

Logan Slade1, Dipsikha Biswas1, Petra C Kienesberger1

  • 1Department of Biochemistry and Molecular Biology, Dalhousie University, Dalhousie Medicine New Brunswick, Saint John, New Brunswick, Canada.

The Journal of Biological Chemistry
|November 13, 2022
PubMed

Insights

Transcription factor EB (TFEB) is crucial for triple-negative breast cancer (TNBC) survival by controlling DNA replication and cell cycle progression. Inhibiting TFEB impairs TNBC cell proliferation and increases sensitivity to Aurora Kinase A inhibitors.

Area of Science:

  • Oncology
  • Molecular Biology
  • Cell Biology

Background:

  • Triple-negative breast cancer (TNBC) presents treatment challenges due to a lack of targeted therapies and high relapse rates.
  • Transcription factor EB (TFEB) has been identified as essential for TNBC survival, regulating DNA repair, apoptosis, and cell cycle pathways.
  • The precise mechanisms by which TFEB influences DNA repair and cell cycle, and their impact on TNBC survival, remain unclear.

Purpose of the Study:

  • To elucidate the specific mechanisms through which TFEB regulates DNA repair and cell cycle pathways in TNBC.
  • To determine if TFEB's effects on these pathways are critical for TNBC cell survival.
  • To investigate the therapeutic potential of targeting TFEB in TNBC.

Main Methods:

  • TFEB knockdown was performed in MDA-MB-231 TNBC cells and MCF10A noncancerous cells.
  • Gene and protein expression related to DNA replication and cell cycle were analyzed.
  • DNA replication was measured using EdU incorporation.
  • Cell cycle progression was assessed after G1/S synchronization.
  • Origin licensing was evaluated in G1 and early S-phase cells.
  • Sensitivity to Aurora Kinase A inhibitors was tested.

Main Results:

  • TFEB knockdown decreased expression of genes and proteins involved in DNA replication and cell cycle progression in TNBC cells.
  • TFEB silencing reduced DNA replication and impaired G1/S phase transition.
  • TFEB knockdown reduced origin licensing in G1 and early S-phase MDA-MB-231 cells.
  • TFEB silencing led to replication stress and G1/S arrest in noncancerous MCF10A cells, but not in TNBC cells.
  • TFEB knockdown sensitized TNBC cells to Aurora Kinase A inhibitors, decreasing cell viability.

Conclusions:

  • TFEB inhibition disrupts cell cycle progression in TNBC by reducing origin licensing and delaying S-phase entry.
  • Targeting TFEB enhances the sensitivity of TNBC cells to Aurora Kinase A inhibitors, offering a potential therapeutic strategy.
  • TFEB silencing has differential effects on cancer and noncancerous cells, causing replication stress and G1/S arrest in the latter.
Keywords:
Aurora kinase ADNA damageDNA replicationMCF-10AMDA-MB-231RB1RNA-SeqStathmin 1TFEBcell cyclegenome stabilityorigin licensingtranscription factortriple negative breast cancer

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