Carcinogenesis, Vol. 21, No. 8, 1513-1519,
August 2000
© 2000 Oxford University Press
Cancer Biology |
Morphodensitometric analysis of protein kinase C ßII expression in rat colon: modulation by diet and relation to in situ cell proliferation and apoptosis
Molecular and Cell Biology Section, Faculty of Nutrition and
1 Department of Statistics, Texas A&M University, College Station, TX 77843, USA
We have recently demonstrated that overexpression of PKC ßII renders transgenic mice more susceptible to carcinogen-induced colonic hyperproliferation and aberrant crypt foci formation. In order to further investigate the ability of PKC ßII to modulate colonocyte cytokinetics, we determined the localization of PKC ßII with respect to cell proliferation and apoptosis along the entire colonic crypt axis following carcinogen and diet manipulation. Rats were provided diets containing either corn oil [containing n-6 polyunsaturated fatty acids (PUFA)] or fish oil (containing n-3 PUFA), cellulose (non-fermentable fiber) or pectin (fermentable fiber) and injected with azoxymethane (AOM) or saline. After 16 weeks, an intermediate time point when no macroscopic tumors are detected, colonic sections were utilized for immunohistochemical image analysis and immunoblotting. Cell proliferation was measured by incorporation of bromodeoxyuridine into DNA and apoptosis by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end-labeling. In the distal colon, PKC ßII staining was localized to the upper portion of the crypt. In comparison, proximal crypts had more (P < 0.05) staining in the lower tertile. AOM enhanced (P < 0.05) PKC ßII expression in all regions of the distal colonic crypt (upper, middle and lower tertiles). There was also an interaction (P < 0.05) between dietary fat and fiber on PKC ßII expression (corn/pectin > fish/cellulose, fish/pectin > corn/cellulose) in all regions of the distal colonic crypt. With respect to colonic cell kinetics, proliferation paralleled the increase in PKC ßII expression in carcinogen-treated animals. In contrast, apoptosis at the lumenal surface was inversely proportional to PKC ßII expression in the upper tertile. These results suggest that an elevation in PKC ßII expression along the crypt axis in the distal colon is linked to enhancement of cell proliferation and suppression of apoptosis, predictive intermediate biomarkers of tumor development. Therefore, select dietary factors may confer protection against colon carcinogenesis in part by blocking carcinogen-induced PKC ßII expression.
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