Identification of small-molecule inhibitors of the colorectal cancer oncogene Krüppel-like factor 5 expression by ultrahigh-throughput screening

Agnieszka B. Bialkowska, Melissa Crisp, Thomas Bannister, Yuanjun He, Sarwat Chowdhury, Stephan Schur̈er, Peter Chase, Timothy Spicer, Franck Madoux, Chenlu Tian, Peter Hodder, Daniel Zaharevitz, Vincent W. Yang

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

The transcription factor Krüppel-like factor 5 (KLF5) is primarily expressed in the proliferative zone of the mammalian intestinal epithelium, where it regulates cell proliferation. Studies showed that inhibition of KLF5 expression reduces proliferation rates in human colorectal cancer cells and intestinal tumor formation in mice. To identify chemical probes that decrease levels of KLF5, we used cell-based ultrahigh-throughput screening (uHTS) to test compounds in the public domain of NIH, the Molecular Libraries Probe Production Centers Network library. The primary screen involved luciferase assays in the DLD-1/pGL4.18hKLF5p cell line, which stably expressed a luciferase reporter driven by the human KLF5 promoter. A cytotoxicity counterscreen was done in the rat intestinal epithelial cell line, IEC-6. We identified 97 KLF5-selective compounds with EC 50 < 10 μmol/L for KLF5 inhibition and EC 50 > 10 μmol/L for IEC-6 cytotoxicity. The two most potent compounds, CIDs (PubChem Compound IDs) 439501 and 5951923, were further characterized on the basis of computational, Western blot, and cell viability analyses. Both of these compounds, and two newly synthesized structural analogs of CID 5951923, significantly reduced endogenous KLF5 protein levels and decreased viability of several colorectal cancer cell lines without any apparent impact on IEC-6 cells. Finally, when tested in the NCI-60 panel of human cancer cell lines, compound CID 5951923 was selectively active against colon cancer cells. Our results show the feasibility of uHTS in identifying novel compounds that inhibit colorectal cancer cell proliferation by targeting KLF5.

Original languageEnglish (US)
Pages (from-to)2043-2051
Number of pages9
JournalMolecular Cancer Therapeutics
Volume10
Issue number11
DOIs
StatePublished - Nov 1 2011

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Oncogenes
Colorectal Neoplasms
Cell Line
Luciferases
Cell Proliferation
Molecular Probes
Public Sector
Intestinal Mucosa
Colonic Neoplasms
Libraries
Neoplasms
Cell Survival
Transcription Factors
Western Blotting
Epithelial Cells
Proteins

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

Cite this

Identification of small-molecule inhibitors of the colorectal cancer oncogene Krüppel-like factor 5 expression by ultrahigh-throughput screening. / Bialkowska, Agnieszka B.; Crisp, Melissa; Bannister, Thomas; He, Yuanjun; Chowdhury, Sarwat; Schur̈er, Stephan; Chase, Peter; Spicer, Timothy; Madoux, Franck; Tian, Chenlu; Hodder, Peter; Zaharevitz, Daniel; Yang, Vincent W.

In: Molecular Cancer Therapeutics, Vol. 10, No. 11, 01.11.2011, p. 2043-2051.

Research output: Contribution to journalArticle

Bialkowska, AB, Crisp, M, Bannister, T, He, Y, Chowdhury, S, Schur̈er, S, Chase, P, Spicer, T, Madoux, F, Tian, C, Hodder, P, Zaharevitz, D & Yang, VW 2011, 'Identification of small-molecule inhibitors of the colorectal cancer oncogene Krüppel-like factor 5 expression by ultrahigh-throughput screening', Molecular Cancer Therapeutics, vol. 10, no. 11, pp. 2043-2051. https://doi.org/10.1158/1535-7163.MCT-11-0550
Bialkowska, Agnieszka B. ; Crisp, Melissa ; Bannister, Thomas ; He, Yuanjun ; Chowdhury, Sarwat ; Schur̈er, Stephan ; Chase, Peter ; Spicer, Timothy ; Madoux, Franck ; Tian, Chenlu ; Hodder, Peter ; Zaharevitz, Daniel ; Yang, Vincent W. / Identification of small-molecule inhibitors of the colorectal cancer oncogene Krüppel-like factor 5 expression by ultrahigh-throughput screening. In: Molecular Cancer Therapeutics. 2011 ; Vol. 10, No. 11. pp. 2043-2051.
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AU - Chowdhury, Sarwat

AU - Schur̈er, Stephan

AU - Chase, Peter

AU - Spicer, Timothy

AU - Madoux, Franck

AU - Tian, Chenlu

AU - Hodder, Peter

AU - Zaharevitz, Daniel

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