Oncotarget

Priority Research Papers:

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

Ruowen Zhang, Jiahui Wu, Sylvain Ferrandon, Katie J. Glowacki, Janet A. Houghton _

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Oncotarget. 2016; 7:80190-80207. https://doi.org/10.18632/oncotarget.13376

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Abstract

Ruowen Zhang1, Jiahui Wu2, Sylvain Ferrandon2, Katie J. Glowacki1 and Janet A. Houghton1

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

2 Lerner Research Institute, Cleveland Clinic, Cleveland, OH, USA

Correspondence to:

Janet A. Houghton, email:

Keywords: GLI, GANT61, transcription, DNA licensing

Received: October 05, 2016 Accepted: November 07, 2016 Published: November 15, 2016

Abstract

The GLI genes are transcription factors and in cancers are oncogenes, aberrantly and constitutively activated. GANT61, a specific GLI inhibitor, has induced extensive cytotoxicity in human models of colon cancer. The FOXM1 promoter was determined to be a transcriptional target of GLI1. In HT29 cells, inhibition of GLI1 binding at the GLI consensus sequence by GANT61 led to inhibited binding of Pol II, the pause-release factors DSIF, NELF and p-TEFb. The formation of R-loops (RNA:DNA hybrids, ssDNA), were reduced by GANT61 at the FOXM1 promoter. Pretreatment of HT29 cells with α-amanitin reduced GANT61-induced γH2AX foci. Co-localization of GLI1 and BrdU foci, inhibited by GANT61, indicated GLI1 and DNA replication to be linked. By co-immunoprecipitation and confocal microscopy, GLI1 co-localized with the DNA licensing factors ORC4, CDT1, and MCM2. Significant co-localization of GLI1 and ORC4 was inhibited by GANT61, and enrichment of ORC4 occurred at the GLI binding site in the FOXM1 promoter. CDT1 was found to be a transcription target of GLI1. Overexpression of CDT1 in HT29 and SW480 cells reduced GANT61-induced cell death, gH2AX foci, and cleavage of caspase-3. Data demonstrate involvement of transcription and of DNA replication licensing factors by non-transcriptional and transcriptional mechanisms in the GLI-dependent mechanism of action of GANT61.

Author Information

Ruowen Zhang

Jiahui Wu

Sylvain Ferrandon

Katie J. Glowacki

Janet A. Houghton
Primary Contact  _


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