Original Article
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World J Gastrointest Oncol. Jul 15, 2014; 6(7): 225-243
Published online Jul 15, 2014. doi: 10.4251/wjgo.v6.i7.225
Novel diet-related mouse model of colon cancer parallels human colon cancer
Anil R Prasad, Shilpa Prasad, Huy Nguyen, Alexander Facista, Cristy Lewis, Beryl Zaitlin, Harris Bernstein, Carol Bernstein
Anil R Prasad, Department of Pathology, Northwest Medical Center, Tucson, AZ 85741, United States
Anil R Prasad, Department of Pathology, College of Medicine, University of Arizona, Tucson, AZ 85724, United States
Shilpa Prasad, College of Arts and Sciences, Boston University, Boston, MA 2215, United States
Huy Nguyen, Alexander Facista, Cristy Lewis, Harris Bernstein, Carol Bernstein, Department of Cellular and Molecular Medicine, College of Medicine, University of Arizona, Tucson, AZ 85724, United States
Beryl Zaitlin, Matrix Solutions Inc., Alberta T2R 0V2, Canada
Author contributions: All authors contributed equally to this work; Bernstein C designed the experiments; Prasad AR performed the pathologic and histologic analysis; Prasad S and Bernstein C collected the digital images; Nguyen H, Facista A and Lewis C performed the immunohistochemistry; Zaitlin B performed the statistical analysis; Prasad AR and Bernstein C drafted the manuscript; and Bernstein H critically revised the manuscript.
Supported by National Institutes of Health, No. 5 R01 CA119087; Arizona Biomedical Research Commission, No. 0803; and Veterans Affairs Merit Review, No. 0142; administered by the Southern Arizona Veterans Affairs Health Care System
Correspondence to: Carol Bernstein, PhD, Department of Cellular and Molecular Medicine, College of Medicine, University of Arizona, 2639 E 4th Street, Tucson, AZ 85716, United States. bernstein324@yahoo.com
Telephone: +1-520-2415260 Fax: +1-520-3240275
Received: October 19, 2013
Revised: April 4, 2014
Accepted: June 18, 2014
Published online: July 15, 2014

AIM: To investigate the close parallels between our novel diet-related mouse model of colon cancer and human colon cancer.

METHODS: Twenty-two wild-type female mice (ages 6-8 wk) were fed the standard control diet (AIN-93G) and an additional 22 female mice (ages 6-8 wk) were fed the control diet supplemented with 0.2% deoxycholic acid [diet + deoxycholic acid (DOC)] for 10 mo. Tumors occurred in the colons of mice fed diet + DOC and showed progression to colon cancer [adenocarcinoma (AC)]. This progression is through the stages of tubular adenoma (TA), TA with high grade dysplasia or adenoma with sessile serrated morphology, intramucosal AC, AC stage T1, and AC stage T2. The mouse tumors were compared to human tumors at the same stages by histopathological analysis. Sections of the small and large intestines of mice and humans were evaluated for glandular architecture, cellular and nuclear morphology including cellular orientation, cellular and nuclear atypia, pleomorphism, mitotic activity, frequency of goblet cells, crypt architecture, ulceration, penetration of crypts through the muscularis mucosa and presence of malignant crypts in the muscularis propria. In addition, preserved colonic tissues from genetically similar male mice, obtained from a prior experiment, were analyzed by immunohistochemistry. The male mice had been fed the control diet or diet + DOC. Four molecular markers were evaluated: 8-OH-dG, DNA repair protein ERCC1, autophagy protein beclin-1 and the nuclear location of beta-catenin in the stem cell region of crypts. Also, male mice fed diet + DOC plus 0.007% chlorogenic acid (diet + DOC + CGA) were evaluated for ERCC1, beclin-1 and nuclear location of beta-catenin.

RESULTS: Humans with high levels of diet-related DOC in their colons are at a substantially increased risk of developing colon cancer. The mice fed diet + DOC had levels of DOC in their colons comparable to that of humans on a high fat diet. The 22 mice without added DOC in their diet had no colonic tumors while 20 of the 22 mice (91%) fed diet + DOC developed colonic tumors. Furthermore, the tumors in 10 of these mice (45% of mice) included an adenocarcinoma. All mice were free of cancers of the small intestine. Histopathologically, the colonic tumor types in the mice were virtually identical to those in humans. In humans, characteristic aberrant changes in molecular markers can be detected both in field defects surrounding cancers (from which cancers arise) and within cancers. In the colonic tissues of mice fed diet + DOC similar changes in biomarkers appeared to occur. Thus, 8-OH-dG was increased, DNA repair protein ERCC1 was decreased, autophagy protein beclin-1 was increased and, in the stem cell region at the base of crypts there was substantial nuclear localization of beta-catenin as well as increased cytoplasmic beta-catenin. However, in mice fed diet + DOC + CGA (with reduced frequency of cancer) and evaluated for ERCC1, beclin-1, and beta-catenin in the stem cell region of crypts, mouse tissue showed amelioration of the aberrancies, suggesting that chlorogenic acid is protective at the molecular level against colon cancer. This is the first diet-related model of colon cancer that closely parallels human progression to colon cancer, both at the histomorphological level as well as in its molecular profile.

CONCLUSION: The diet-related mouse model of colon cancer parallels progression to colon cancer in humans, and should be uniquely useful in model studies of prevention and therapeutics.

Keywords: Diet, Deoxycholate, Mouse model, Colon cancer, Histology, Chlorogenic acid, 8-OH-dG, Beclin 1, Beta-catenin

Core tip: Mouse models of colon carcinogenesis are essential as platforms for trials of prevention and therapy. However, most previous rodent models of colon carcinogenesis lack an invasive phenotype and/or do not share several significant genetic events and histopathological features of human colon cancer. This new diet-related mouse model of colon cancer is unique in being closely parallel to human progression to sporadic colon cancer by measures of its histomorphology and its molecular profile. It also has a natural basis, using dietary deoxycholic acid, long thought to be a central causative agent in colon carcinogenesis.