Journal of Biological Chemistry
Volume 289, Issue 36, 5 September 2014, Pages 25306-25316
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Gene Regulation
The Intestinal Epithelial Cell Differentiation Marker Intestinal Alkaline Phosphatase (ALPi) Is Selectively Induced by Histone Deacetylase Inhibitors (HDACi) in Colon Cancer Cells in a Kruppel-like Factor 5 (KLF5)-dependent Manner*

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The histone deacetylase inhibitor (HDACi) sodium butyrate promotes differentiation of colon cancer cells as evidenced by induced expression and enzyme activity of the differentiation marker intestinal alkaline phosphatase (ALPi). Screening of a panel of 33 colon cancer cell lines identified cell lines sensitive (42%) and resistant (58%) to butyrate induction of ALP activity. This differential sensitivity was similarly evident following treatment with the structurally distinct HDACi, MS-275. Resistant cell lines were significantly enriched for those harboring the CpG island methylator phenotype (p = 0.036, Chi square test), and resistant cell lines harbored methylation of the ALPi promoter, particularly of a CpG site within a critical KLF/Sp regulatory element required for butyrate induction of ALPi promoter activity. However, butyrate induction of an exogenous ALPi promoter-reporter paralleled up-regulation of endogenous ALPi expression across the cell lines, suggesting the presence or absence of a key transcriptional regulator is the major determinant of ALPi induction. Through microarray profiling of sensitive and resistant cell lines, we identified KLF5 to be both basally more highly expressed as well as preferentially induced by butyrate in sensitive cell lines. KLF5 overexpression induced ALPi promoter-reporter activity in resistant cell lines, KLF5 knockdown attenuated butyrate induction of ALPi expression in sensitive lines, and butyrate selectively enhanced KLF5 binding to the ALPi promoter in sensitive cells. These findings demonstrate that butyrate induction of the cell differentiation marker ALPi is mediated through KLF5 and identifies subsets of colon cancer cell lines responsive and refractory to this effect.

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*

This work was supported in part by the Operational Infrastructure Support Program (Victorian government, Australia), Spanish Ministry for Economy and Competitiveness Grants CP05/00256, TRA2009-0093, SAF2008-00789, PI12/03112, and PI12/01095, the Association for International Cancer Research Grant AICR13-0245, Agència de Gestió d'Ajuts Universitaris i de Recerca SGR 157 (to D. A.), and Australian Research Council Future Fellowship FT0992234 and National Health and Medical Research Council Senior Research Fellowship 1046092 (to J. M. M.).

This article contains supplemental Table 1.

1

Both authors contributed equally to this work.

2

Supported by a fellowship from CONACyT Mexico (186310).

4

The abbreviations used are:

    HDAC

    histone deacetylase

    ALPi

    intestinal alkaline phosphatase

    KLF

    Kruppel-like factor

    CIMP

    CpG Island methylator phenotype.