Cancer-Predicting Gene Expression Changes in Colonic Mucosa of Western Diet Fed Mlh 1 ( + /-) Mice Pussila , Marjaana 2013-1008

Colorectal cancer (CRC) is the second most common cause of cancer-related deaths in the Western world and interactions between genetic and environmental factors, including diet, are suggested to play a critical role in its etiology. We conducted a long-term feeding experiment in the mouse to address gene expression and methylation changes arising in histologically normal colonic mucosa as putative cancer-predisposing events available for early detection. The expression of 94 growth-regulatory genes previously linked to human CRC was studied at two time points (5 weeks and 12 months of age) in the heterozygote Mlh1+/mice, an animal model for human Lynch syndrome (LS), and wild type Mlh1+/+ littermates, fed by either Western-style (WD) or AIN-93G control diet. In mice fed with WD, proximal colon mucosa, the predominant site of cancer formation in LS, exhibited a significant expression decrease in tumor suppressor genes, Dkk1, Hoxd1, Slc5a8, and Socs1, the latter two only in the Mlh1+/mice. Reduced mRNA expression was accompanied by increased promoter methylation of the respective genes. The strongest expression decrease (7.3 fold) together with a significant increase in its promoter methylation was seen in Dkk1, an antagonist of the canonical Wnt signaling pathway. Furthermore, the inactivation of Dkk1 seems to predispose to neoplasias in the proximal colon. This and the fact that Mlh1 which showed only modest methylation was still expressed in both Mlh1+/and Mlh1+/+ mice indicate that the expression decreases and the inactivation of Dkk1 in particular is a prominent early marker for colon oncogenesis. Citation: Pussila M, Sarantaus L, Dermadi Bebek D, Valo S, Reyhani N, et al. (2013) Cancer-Predicting Gene Expression Changes in Colonic Mucosa of Western Diet Fed Mlh1+/Mice. PLoS ONE 8(10): e76865. doi:10.1371/journal.pone.0076865 Editor: Ajay Goel, Baylor University Medical Center, United States of America Received May 22, 2013; Accepted August 28, 2013; Published October 8, 2013 Copyright: © 2013 Pussila et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Funding: This work was financially supported by the grants from European Research Council (2008-AdG-232635), The Sigrid Juselius Foundation (http:// www.sigridjuselius.fi/foundation), Finnish Cancer Organizations (http://www.cancer.fi/en/), The Academy of Finland (http://www.aka.fi/eng), and Biocentrum Helsinki (http://www.helsinki.fi/biocentrum/). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Competing interests: The authors have declared that no competing interests exist. * E-mail: minna.nystrom@helsinki.fi ☯ These authors contributed equally to this work.