Paneth cell marker expression in intestinal villi and colon crypts characterizes dietary induced risk for mouse sporadic intestinal cancer - PubMed (original) (raw)

Paneth cell marker expression in intestinal villi and colon crypts characterizes dietary induced risk for mouse sporadic intestinal cancer

Donghai Wang et al. Proc Natl Acad Sci U S A. 2011.

Abstract

Nutritional and genetic risk factors for intestinal tumors are additive on mouse tumor phenotype, establishing that diet and genetic factors impact risk by distinct combinatorial mechanisms. In a mouse model of dietary-induced sporadic small and large intestinal cancer in WT mice in which tumor etiology, lag, incidence, and frequency reflect >90% of intestinal cancer in Western societies, dietary-induced risk altered gene expression profiles predominantly in villus cells of the histologically normal mucosa, in contrast to targeting of crypt cells by inheritance of an Apc(1638N) allele or homozygous inactivation of p21(Waf1/cip1), and profiles induced by each risk factor were distinct at the gene or functional group level. The dietary-induced changes in villus cells encompassed ectopic expression of Paneth cell markers (a lineage normally confined to the bottom of small intestinal crypts), elevated expression of the Wnt receptor Fzd5 and of EphB2 (genes necessary for Paneth cell differentiation and localization to the crypt bottom), and increased Wnt signaling in villus cells. Ectopic elevation of these markers was also present in the colon crypts, which are also sites of sporadic tumors in the nutritional model. Elevating dietary vitamin D(3) and calcium, which prevents tumor development, abrogated these changes in the villus and colon cells. Thus, common intestinal cancer driven by diet involves mechanisms of tumor development distinct from those mechanisms that cause tumors induced by the rare inheritance of a mutant adenomatous polyposis coli (Apc) allele. This is fundamental for understanding how common sporadic tumors arise and in evaluating relative risk in the population.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Fig. 1.

Fig. 1.

Unsupervised clustering of Affymetrix expression data for crypt and villus cells based on the mean of each probe for the four mice of each group. Expression profiles of villus and crypt cells are distinctly independent of dietary/genetic risk.

Fig. S1

has details of data generation and analyses.

Fig. 2.

Fig. 2.

Limited overlap of gene expression changes for each dietary and genetic risk group in both villus and crypt cells.

Fig. 3.

Fig. 3.

Level of expression of Paneth cell markers, Fzd5, and Ephb2 in villus and crypt cells of mice maintained on AIN76A, NWD1, and NWD2 from weaning for 1 y. Expression levels determined by qRT-PCR and normalized to GAPDH expression, determined similarly.

Fig. 4.

Fig. 4.

Immunohistochemical detection of the Paneth cell marker defensin in the intestinal mucosa of mice maintained on AIN76A, NWD1, or NWD2 for 1 y from weaning. Positive staining is brown and localized to Paneth cells in the crypt for mice fed AIN76A or NWD2, but also, it is present in the apical region of normal-appearing enterocytes in mice fed NWD1.

Fig. 5.

Fig. 5.

Immunohistochemical detection of the Wnt receptor Fzd 5 in mice fed AIN76A, NWD1, or NWD2 for 1 y from weaning.

Fig. 6.

Fig. 6.

Detection of Wnt signaling by assay of β-galactosidase activity (blue) in frozen sections of the mucosa of C57BL/6 mice that harbor a lacZ transgene driven by a minimal fos promoter under regulation of three tandem Tcf4 binding sites. In this case, mice were fed the three diets from weaning for 3 mo.

Fig. 7.

Fig. 7.

Immunohistochemical detection of defensin, lysozyme, Fzd5, and β-galactosidase in the colon of mice fed AIN76A, NWD1, or NWD2 for 1 y from weaning (3 mo for beta-galactosidase).

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