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Targeting GLI by GANT61 involves mechanisms dependent on inhibition of both transcription and DNA licensing

Ruowen Zhang1, Jiahui Wu2, Sylvain Ferrandon2

  • 1Department of Oncology, Division of Drug Discovery, Southern Research, Birmingham, AL, USA.

Oncotarget
|November 19, 2016
PubMed

Insights

The GLI inhibitor GANT61 halts colon cancer cell growth by blocking GLI1

Area of Science:

  • Oncology
  • Molecular Biology
  • Cancer Genetics

Background:

  • GLI genes are transcription factors and oncogenes in cancer.
  • Aberrant GLI activation drives tumor progression.
  • GANT61 is a GLI inhibitor with demonstrated cytotoxicity in colon cancer models.

Purpose of the Study:

  • To elucidate the molecular mechanisms of GANT61 action.
  • To investigate the interplay between GLI1, transcription, and DNA replication.
  • To identify GLI1 transcriptional targets involved in GANT61's efficacy.

Main Methods:

  • GLI inhibitor GANT61 treatment in HT29 colon cancer cells.
  • Chromatin immunoprecipitation to assess GLI1 binding and Pol II recruitment.
  • Analysis of R-loop formation, DNA replication markers (BrdU), and DNA licensing factors (ORC4, CDT1, MCM2).
  • Co-immunoprecipitation and confocal microscopy to study protein interactions.
  • Assessment of cell death markers (γH2AX, caspase-3 cleavage) and effect of CDT1 overexpression.

Main Results:

  • GANT61 inhibited GLI1 binding, Pol II recruitment, and R-loop formation at the FOXM1 promoter.
  • GANT61 disrupted the link between GLI1 and DNA replication.
  • GLI1 was found to co-localize with DNA licensing factors ORC4, CDT1, and MCM2.
  • CDT1 was identified as a GLI1 transcriptional target, and its overexpression reduced GANT61-induced cell death.
  • GANT61's mechanism involves both transcriptional and non-transcriptional effects on DNA replication licensing factors.

Conclusions:

  • GANT61 exerts its anti-cancer effects by inhibiting GLI1-mediated transcription and disrupting DNA replication licensing.
  • The findings reveal a novel mechanism of action for GLI inhibitors in colon cancer.
  • Targeting GLI1 and its downstream effectors like CDT1 holds therapeutic potential for colon cancer treatment.
Keywords:
DNA licensingGANT61GLItranscription

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