Dietary energy restriction, in part through glucocorticoid hormones, mediates the impact of 12-O-tetradecanoylphorbol-13-acetate on jun D and fra-1 in Sencar mouse epidermis.

Dietary energy restriction (DER, 40% calorie reduction from fat and carbohydrate) inhibited mouse skin carcinogenesis and decreased 12-O-tetradecanoyl-13-phorbol acetate (TPA)-induced activator protein-1 (AP-1):DNA binding previously. This study measured protein levels of c-jun, jun B, jun D, c-fos, fra-1, and fra-2 and examined their contribution to AP-1:DNA binding by electrophoretic mobility shift assay (EMSA) with supershift analysis in the epidermis of control and DER Sencar mice exposed to TPA. TPA significantly increased c-jun, jun B, c-fos, fra-1, and fra-2 and decreased jun D within 3-6 h after treatment. AP-1:DNA binding reached a maximum 2.5-fold induction over controls 4 h after TPA treatment and antibodies to jun B, jun D, and fra-2 in the EMSA binding reaction resulted in supershifts in both acetone- and TPA-treated mice 1-6 h after treatment. The effect of corticosterone (CCS) and DER on the AP-1 proteins and on the composition of the AP-1:DNA complex was measured in adrenalectomized (adx) mice. DER reduced the TPA impact on jun D and enhanced the induction of fra-1. In addition, CCS-supplemented groups had significantly lower jun D and higher fra-2 than adx groups and sham groups. While sham animals treated with either acetone or TPA contained jun B, jun D, and fra-2 proteins in the AP-1:DNA complex by supershift analysis, fra-2 was no longer seen in adx DER animals. In summary, our study supports potential roles for jun D, jun B, and fra-1 in the DER regulation of AP-1 function in the Sencar mouse skin carcinogenesis model.

Effect of dietary apigenin on colonic ornithine decarboxylase activity, aberrant crypt foci formation, and tumorigenesis in different experimental models.

The efficacy of dietary apigenin, a dietary flavonoid, in colon cancer prevention was investigated by evaluating the inhibition of the ornithine decarboxylase (ODC) activity and the formation of aberrant crypt foci (ACF) and by studying the ability of apigenin to block colon carcinogenesis in two mouse models. First, the activity of ODC was measured in colon cancer cells (Caco-2) and in the colon epithelium of CF-1 mice. Apigenin at 10 and 30 muM significantly inhibited the ODC activity of Caco-2 cells by 26% and 57%, respectively. Colonic ODC activity in CF-1 mice was reduced with 0.1% dietary apigenin by 42% compared with the control, but this difference was not statistically significant. Second, ACF formation was evaluated in azoxymethane (AOM)-induced CF-1 mice. Female CF-1 mice at 6 wk of age were i.p. injected with 5 mg/kg body weight (BW) AOM once to induce ACF. ACF formation in CF-1 mice was reduced by 50% (P < 0.05) with 0.1% dietary apigenin fed for 6 wk when compared with the control. Dietary apigenin inhibited ACF only in the distal region of the CF-1 mouse colon. Finally, tumorigenesis studies were conducted using two different mouse models: AOM-induced CF-1 mice and Min mice with mutant adenomatous polyposis coli (APC) gene. Female CF-1 mice at 6 wk of age were i.p. injected with 10 mg/kg BW AOM weekly for 6 (AOM Study I) or 4 (AOM Study II) wk to induce tumors. CF-1 mice were fed diets containing 0.025% or 0.1% apigenin for 23-25 wk. Female Min mice were fed diets for 10 wk beginning at 5 wk of age. In two AOM-treated mouse colon tumor studies 0.025% and 0.1% dietary apigenin modestly reduced tumors in the group fed 0.025% apigenin (25% incidence in comparison with 65% in the controls) in a non-dose response manner. Apigenin failed to inhibit adenoma formation in the Min mouse study. These results suggest that dietary apigenin showed promise in cancer prevention by reducing the ODC activity and ACF formation, however, clear evidence of cancer prevention was not obtained in mouse tumor studies. Further investigation of the potential chemopreventive effect of apigenin in carcinogenesis is warranted.

Prevention of mouse skin tumor promotion by dietary energy restriction requires an intact adrenal gland and glucocorticoid supplementation restores inhibition.

Our laboratory has demonstrated in the previous studies that dietary energy restriction (DER) inhibited the promotion of skin tumorigenesis and others have found that adrenalectomy may reverse that inhibition. The purpose of the research reported here was to determine if circulating corticosterone (CCS) may be the adrenal hormone responsible for DER prevention of skin carcinogenesis. Female SENCAR mice were initiated with 7,12-dimethylbenzanthracene (DMBA) and promoted with 12-O-tetradecanoylphorbol-13-acetate (TPA) in either sham-operated or adrenalectomized (ADX) mice fed ad libitum (AL) or energy restricted diets. DER was 60% of the AL calorie intake with the removal of energy from fat and carbohydrate. CCS, the main glucocorticoid hormone secreted by the murine adrenal gland, was added to the drinking water of AL/ADX and DER/ADX groups to determine the role of CCS in the DER inhibition of tumor development. In sham-operated groups, DER compared with AL-fed mice experienced significantly decreased papilloma incidence and multiplicity (P < 0.0001). ADX did not alter papilloma incidence or multiplicity in AL-fed mice but ADX partially reversed the inhibition of papilloma multiplicity and incidence in DER mice. CCS supplementation to both DER/ADX and AL/ADX mice resulted in reduced papilloma incidence and multiplicity. In DER/ADX mice, CCS dramatically reduced papilloma rates while in AL/ADX mice CCS reduced the papilloma rates to those seen in the DER sham group. DER significantly reduced carcinoma multiplicity mean counts per effective animal (P < 0.0001) compared with AL-fed groups in sham and ADX/CCS groups. DER/ADX mice lost the carcinoma multiplicity protection seen in sham/DER mice. CCS treatment of ADX mice significantly decreased total carcinoma (in situ and invasive) incidence rates per effective animal (P < 0.0003). ADX followed by CCS treatment in the DER mice resulted in the lowest carcinoma incidence and multiplicity. Thus, DER-inhibition of skin tumorigenesis was mediated at least in part through CCS. However, CCS was more effective in preventing papillomas and carcinomas in DER/ADX mice than in AL/ADX mice, suggesting that other factors may also be involved in the DER prevention of tumor formation.