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High incidence of LRAT promoter hypermethylation in colorectal cancer correlates with tumor stage

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Abstract

Lecithin:retinol acyltransferase (LRAT) is a major enzyme involved in vitamin A/retinol metabolism, which regulates various physiological processes like cell proliferation and differentiation. LRAT expression is reduced in numerous cancers, yet the underlying mechanisms have remained undefined. We hypothesized that methylation silencing may contribute to decreased LRAT gene expression in colorectal cancer (CRC). LRAT hypermethylation status was analyzed in five CRC cell lines, 167 colorectal tumors, and 69 adjacent normal colonic mucosae, using a quantitative bisulfite/PCR/LDR/Universal Array assay. LRAT transcription levels were determined by real-time RT-PCR in a subset of tumors and matched normal tissues and in CRC cell lines that were treated with a demethylating agent, 5-aza-2′-deoxycytidine. The incidence of LRAT hypermethylation was significantly higher in colorectal tumors than in adjacent normal mucosae (p = 0.0025). Aberrant methylation occurred in 51 % of microsatellite-stable CRCs, in 84 % of microsatellite-unstable CRCs, and in 12 out of 13 colonic polyps. The number of hypermethylated LRAT events was inversely correlated with CRC stage (p < 0.0001). Importantly, LRAT hypermethylation was associated with decreased mRNA level in CRC clinical specimens, and demethylation treatment resulted in LRAT transcriptional reactivation. Our data support the idea that LRAT promoter hypermethylation associates with LRAT gene expression in CRC. The higher frequency of LRAT hypermethylation in colonic polyps and early-stage CRCs indicates that it may occur early in malignant progression.

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Abbreviations

CIMP:

CpG island methylator phenotype

MSP:

Methylation-specific PCR

LDR:

Ligase detection reaction

LRAT:

Lecithin:retinol acyltransferase

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Acknowledgments

The authors thank WeiJi Shi for the advice in statistical analysis. We also thank Owen Parker for insightful discussion and critical reading of the manuscript, Ludwig Institute for Cancer Research/Conrad N. Hilton Foundation joint Hilton-Ludwig Cancer Metastasis Initiative, and the Gilbert Family Foundation.

Conflict of interest

None.

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Authors and Affiliations

  1. Department of Laboratory Medicine, Cleveland Clinic, Cleveland, OH, 44195, USA

    Yu-Wei Cheng

  2. Department of Neurology and Neuroscience, Icahn School of Medicine at Mount Sinai, New York, NY, 10029, USA

    Hanna Pincas

  3. Department of Microbiology and Immunology, Weill Medical College of Cornell University, New York, NY, 10021, USA

    Jianmin Huang & Francis Barany

  4. Rutgers-Cancer Institute of New Jersey, New Brunswick, NJ, 08903-0019, USA

    Emmanuel Zachariah

  5. Department of Surgery, Colorectal Surgery Service, Memorial Sloan-Kettering Cancer Center, New York, NY, 10021, USA

    Zhaoshi Zeng & Philip Paty

  6. PennState University Hershey Medical Center, Hershey, PA, 17033, USA

    Daniel A. Notterman

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  1. Yu-Wei Cheng
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Correspondence to Yu-Wei Cheng.

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Yu-Wei Cheng and Hanna Pincas have contributed equally to this work.

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Cheng, YW., Pincas, H., Huang, J. et al. High incidence of LRAT promoter hypermethylation in colorectal cancer correlates with tumor stage. Med Oncol 31, 254 (2014). https://doi.org/10.1007/s12032-014-0254-7

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