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Epigenetic control of HNF-4α in colon carcinoma cells affects MUC4 expression and malignancy

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Abstract

Background

We previously found that enhanced expression of hepatocyte nuclear factor 4α (HNF-4α) is associated with hyper-proliferation of colon carcinoma cells. Here, the effect of histone deacetylase (HDAC) inhibitors on proliferation and the expression of HNF-4α and its downstream target genes were assessed in HM7, LS174T, HT29 and Caco-2 colon carcinoma cell lines.

Results

HNF-4α expression was found to vary in the different colon carcinoma cell lines tested, being highest in HM7. Additionally, a direct correlation with proliferation was observed. In HM7 cells, the weak HDAC inhibitor butyrate significantly inhibited the transcription of HNF-4α, its downstream target gene MUC4, and genes associated with proliferation, including the proliferating cell nuclear antigen gene PCNA. siRNA-mediated silencing of HNF-4α exerted an effect similar to butyrate on HM7 cell proliferation. The stronger HDAC inhibitor trichostatin A (TSA) exerted an effect similar to that of siRNA-mediated HNF-4α silencing and, concomitantly, inhibited the expression of the transcription factor gene SP1. Also, siRNA-mediated silencing of HDAC3 and HDAC4 reduced HNF-4α expression. Chromatin immunoprecipitation (ChIP) assays revealed that TSA induces hyperacetylation of histones H3 and H4 and, concomitantly, inhibits SP1 binding to the HNF-4α promoter. Subsequent electromobility shift assays supported these latter findings.

Conclusions

HNF-4α transcriptional expression and activity are tightly controlled by epigenetic mechanisms. HDAC inhibitor targeting of HNF-4α may serve as an effective treatment for advanced colon carcinomas, since downstream cancer-associated target genes such as MUC4 are significantly down-regulated by this treatment.

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Acknowledgments

This work was supported by the Israel Science Foundation; Grant number: 134/06.

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  1. Institute of Biochemistry, Food Science and Nutrition, The Robert H. Smith Faculty of Agriculture, Food and Environment, The Hebrew University of Jerusalem, P.O. Box 12, Rehovot, 76100, Israel

    Anna Algamas-Dimantov, Einav Yehuda-Shnaidman, Irena Peri & Betty Schwartz

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  1. Anna Algamas-Dimantov
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  2. Einav Yehuda-Shnaidman
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Correspondence to Betty Schwartz.

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Supplemental Fig. 1

Effect of butyrate on HNF-4α and MUC4 expression in HM7, LS174T, HT29 and Caco-2 cell lines. (A) Representative western blot analysis of HNF-4α expression in HM7, LS174T, HT29 and Caco-2. Effect of HDACI butyrate (5 mM) treatment on HNF-4α expression in these cell lines. (B) HNF-4α expression normalized to β-actin, mean ± SEM of six independent determinations for each cell line. *,Significantly different from nontreated HM7 cell line at P < 0.01. (JPEG 147 kb)

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Algamas-Dimantov, A., Yehuda-Shnaidman, E., Peri, I. et al. Epigenetic control of HNF-4α in colon carcinoma cells affects MUC4 expression and malignancy. Cell Oncol. 36, 155–167 (2013). https://doi.org/10.1007/s13402-012-0123-3

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