Selenium (Se) is known as an antioxidant mineral and heme iron is a major source for iron intake which can promote carcinogenesis in the body. This study was to investigate the effect of Se on heme-aggravated colon carcinogenesis in mice. Three experimental groups included control [normal diet + AOM (10 mg/kg body weight in saline)/DSS (2% in the drinking water)], [AOM/DSS + hemin (534 mg/kg body weight in CMC)], and [AOM/DSS + hemin + Se (2.82 mg/kg diet in CMC)] groups. Colonic mucosa were stained with 0.3% methylene blue and the colonic polyps, aberrant crypt (AC) and aberrant crypt foci (ACF) were counted. Lipid peroxidation in liver was evaluated by the thiobarbituric acid-reactive substances (TBARS) assay. The number of polyps in the hemin + Se group was 31.6% lower than that in the control group, and 41.4% lower than that in the hemin group. The number of AC in the hemin + Se group was 42.8% lower than that in the control group, and 49.1% lower than that in the hemin group. The number of ACF in the hemin + Se group was 49.0% lower than that in the control group, 45.7% than that in the hemin group. Hepatic TBARS level in the hemin + Se group was significantly low compared with the control group or the hemin group (p<0.05). These findings suggest that Se treatment may be protective against colon carcinogenesis promoted by a high heme-containing diet.

Colorectal cancer causes the most cancer-associated death worldwide, having a high cancer incidence. Pectin is a complex polysaccharide present in various fruits, emerging as an anti-carcinogenic candidate. Although pectin has a suppressive capacity for colon carcinogenesis, the effect of reactive oxygen species (ROS) generation and colonic aberrant foci formation in the colon carcinogenesis mouse model remains unclear. Therefore, this study investigates the regulatory effect of pectin supplementation on colon carcinogenesis induced by azoxymethane (AOM) and dextran sodium sulfate (DSS) in mice. In an animal experiment, thirty male institute for cancer research (ICR) mice were divided into two experimental groups; AOM/DSS (control group) and AOM/DSS + pectin (5% in drinking water). Furthermore, the number of aberrant crypt foci (ACF) and aberrant crypt (AC) on colonic mucosa were counted, and thiobarbituric acid-reactive substances (TBARS) assay was performed to estimate lipid peroxidation in feces. Pectin treatment significantly decreased the number of ACF and AC per colon compared with the control. Additionally, fecal TBARS level in the pectin group was significantly lower than those in the control group. Conclusively, these findings indicate that pectin-inhibited hyperplastic alteration and oxidative stress suppress colitis-associated colon carcinogenesis.

Colon cancer has been considered a leading cause of cancer-associated death. Folic acid is a vitamin necessary for cellular physiological functions and cell viability. However, the association between folic acid intake and colon cancer has been examined in several prospective cohort studies are controversial. This study investigated the effects of folate intake on colon carcinogenesis and oxidative stress in an azoxymethane (AOM)/dextran sodium sulfate (DSS) institute for cancer research (ICR) mouse model. Thirty male ICR mice (5 weeks old) were divided into the control group and the experimental group supplied 0.03% folic acid via drinking water (50 mL/week/mouse) for 6 weeks. To induce colonic pre-neoplastic lesions, the animals were subcutaneously injected three times weekly with AOM (10 mg/kg body weight), followed by 2% DSS in drinking water for a week. Folic acid supplementation significantly suppressed the total number of aberrant crypt foci and aberrant crypts. Histological image data showed that folic acid supplementation attenuated neoplastic change. In addition, we measured the thiobarbituric acid reactive substances concentration of dry feces samples to identify the effect of folic acid on reactive oxygen accumulation. The folic acid supplementation group had reduced reactive oxygen species levels in dry feces compared to the control group. In conclusion, these findings indicate that folic acid suppresses colon carcinogenesis and oxidative stress in an AOM/DSS mouse model.

Colon cancer is known as the third most widespread cancer in the world. The interaction of heme-iron and ascorbic acid (AA) in colon carcinogenesis is not evident. Hemin (ferric chloride heme) is an iron-containing porphyrin with chlorine that can be formed from a heme group. The purpose of this study was to investigate the protective effect of AA on the formation of pre-neoplastic lesions induced by azoxymethane (AOM)/dextran sodium sulfate (DSS) plus hemin in mice. Forty-five ICR male mice were divided into three experimental groups; AOM/ DSS treatment (control group), hemin (2 g hemin/kg of b.w.), hemin + AA (1.0% in drinking water). The mice had three s.c. injections (0–2nd weeks of the experiment) of AOM (10 mg/kg b.w.) weekly and 2% DSS as drinking water for the next one week and the animals fed on AIN-76A purified rodent diet for 6 weeks. The numbers of aberrant crypt foci (ACF) and aberrant crypts (ACs) in colonic mucosa were counted after methylene blue staining. Lipid peroxidation in feces was measured by the thiobarbituric acid-reactive substances (TBARS) assay. The numbers of ACF and ACs per colon significantly increased in Hemin group compared to the control group. However, the numbers of ACF and ACs per colon notably decreased in hemin + AA group compared to the control group or hemin group (p<0.05). In feces, the TBARS value of hemin group was higher than the control group (p<0.01). The TBARS value of hemin + AA group was slightly decreased compared to Hemin group. These results indicate that hemin can promote the experimental colon carcinogenesis in ICR mice. On the other hand, additional supplement of AA via drinking water has a protective effect against the colon carcinogenesis. The related mechanisms need to be illustrated by further studies in future.

Globally, colon cancer is increased gradually and known as one of the major causes of cancer death. Stevia, a substitute of sugar, is known to have many components including alpha-tocopherol and anthocyanin etc, as antioxidants. This study's purpose is to investigate whether stevia plant extract can have a protective effect against colon carcinogenesis induced by azoxymethane (AOM) and dextran sodium sulfate (DSS) in mice. Total 30 male ICR mice were divided into 2 groups; AOM/DSS treatment (control group), AOM/DSS + stevia extract (0.5%, in drinking water). After acclimation for 1 week, five weeks old mice received three intraperitoneal AOM (10 mg/kg b.w.) injections weekly for 3 weeks (0–2nd weeks of the experiment) and 2% DSS as drinking water for the next one week. AIN-76A purified rodent diet and 0.5% stevia extract water were supplied to the animals for 6 weeks. The colons of mice were collected and the number of aberrant crypt foci (ACF) and aberrant crypts (ACs) in colonic mucosa were counted after staining with methylene blue. Malondialdehyde (MDA) concentration in feces were determined. The numbers of ACF and ACs were significantly (p<0.01) decreased in stevia-treated group compared with the control group. The MDA concentration in feces was also significantly (p<0.01) decreased in stevia-treated group compared with the control group. In histopathology of colonic epithelium, hyperplasia of colonic epithelium was less observed in steviatreated group. These results indicate that stevia has a protective effect against colon carcinogenesis induced by AOM/DSS in mice and further study needs to illustrate the protective mechanisms.

Excessive intake of red meat has been associated with colon carcinogenesis. The effect of hemin and zinc on colon carcinogenesis was investigated in male ICR mice. After acclimation for 1 week, five-week-old mice received three s.c. injections (0-2nd weeks of the experiment) of azoxymethane (AOM; 10 mg/kg b.w.) weekly and were treated by 2% dextran sodium sulfate (DSS) in drinking water for the next 1 week to induce aberrant crypt foci (ACF). Mice were fed on AIN-76A purified rodent diet for 6 weeks. There were three experimental groups; control, hemin, and hemin + zinc groups. The daily oral doses of hemin and zinc were 534 mg/kg and 55 mg/kg b.w., respectively. After staining colonic mucosa with 0.2% methylene blue, the ACF, aberrant crypts (AC), and polyps were counted. Lipid peroxidation in liver was evaluated by thiobarbituric acid-reactive substances (TBARS) assay. The numbers of AC, ACF, large ACF (i.e., ≥4 AC/ACF), and polyps in the hemin group were higher than those in the control group. In hemin + zinc group, the numbers of AC, ACF, large ACF were significantly lower compared to the hemin or control groups (p < 0.05), and the number of polyps was also significantly lower compared to that in the hemin group (p < 0.05). The TBARS level in the livers of the hemin + zinc group was significantly lower than that of the hemin group (p < 0.05). These results suggest that hemin enhances colon carcinogenesis and that co-treatment with zinc can protect against the induction of colon carcinogenesis.

Excessive iron can promote the production of free radicals, thereby leading to harmful effects on cancer and aging. Ascorbic acid is not only an antioxidant but also a co-factor of iron absorption. The effect of iron-overload with ascorbic acid on experimental colon carcinogenesis was investigated in male ICR mice. Animals were treated weekly with azoxymethane (AOM, 10 mg/kg b.w.) at 0, 1, and 2 week and then drunk 2% dextran sodium sulfate (DSS)-containing water for the next 1 week. There were four experimental groups: carboxymethylcellulose (CMC) alone (control), CMC + ascorbic acid (AA), CMC + Fe, CMC + Fe + AA. The animals fed on AIN-76A purified rodent diet for six weeks. AA or Fe2O3 at the dose of 450 mg/kg b.w. were daily and orally treated for 6 weeks. The colonic mucosa was stained with methylene blue and then aberrant crypt foci (ACF) and polyps were counted. Thiobarbituric acid-reactive substances (TBARS) in serum and liver were determined. Iron concentration in liver was measured by inductively coupled plasma spectrophotometer. Fe-overload with AA strongly increased liver iron contents compared to control or Fe group (p<0.05). There were no significant differences in the number of ACF or polyps among all groups, although ironoverloaded groups had slightly higher numbers compared with the control or AA group. TBARS values in the liver were increased in the iron-overloaded groups compared to control and AA only group (p<0.05), but serum TBARS values were not changed. These results indicate that the excessive iron treatment did not affect the experimental colon carcinogenesis regardless of presence of AA in mice.

Colorectal cancer is one of the most common types of cancer in men and women who consume a Western diet. We investigated the inhibitory effect of selenium (sodium selenite, Na2SeO3) and selenium nanoparticles (nano-Se) on experimental colon carcinogenesis in ICR mice. After a 1-week acclimation, 6-week-old mice received three intraperitoneal (i.p.) injections (experimental week 0-2) of azoxymethane (AOM, 10 mg/kg body weight, b.w.), followed by 2% dextran sodium sulfate (DSS)-containing drinking water for the next 1 week. The three groups (10 mice/group) were orally administered either distilled water (control), selenium (1.7 ppm), or nano-Se (1.7 ppm) daily for 8 weeks. The numbers of aberrant crypt foci (ACF), aberrant crypt (AC), and tumorous lesions were measured in colonic mucosa. Se and nano-Se treatments significantly decreased the number of ACF, AC, and tumorous lesions compared with the control. However, there was no significant difference between the selenium and nano-Se groups. The glutathione peroxidase (GSH-Px) activity in the liver and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity in serum, were high in the selenium and nano-Se groups, while thiobarbituric acid reactive substance (TBARS) level was low in both Se and nano-Se groups when compared with that in the control group. These findings indicate that selenium and nano-Se showed similar protective effects against colon carcinogenesis by inhibiting the development of ACF and tumorous lesions in mice.

Calcium exerts antiproliferative effects on cellular targets through the promotion of differentiation and apoptosis. We investigated the influence of calcium on the formation of colonic aberrant crypt foci (ACFs), which were induced by exposure to azoxymethane (AOM) followed by dextran sodium sulfate (DSS), in ICR mice. Six-week-old ICR mice received 3 (weeks 0–2) intraperitoneal injections of AOM (10 mg/kg BW), followed by treatment with 2% DSS via drinking water for a week to induce preneoplastic lesions. The mice were then divided into 3 groups: the control (AOM/DSS), AOM/DSS + 1.0% Ca, and AOM/DSS + 2.0% Ca groups. Calcium (1.0 or 2.0%) was administered via drinking water for 12 weeks. After sacrificing the mice, the total numbers of aberrant crypts (ACs) and ACFs were measured in the colonic mucosa after methylene blue staining. The control group displayed 11.58 ± 2.43 ACFs/colon, which were composed of a total of 30.42 ± 5.18 ACs/colon. The number of ACFs with more than 3 ACs, which are likely to progress to colon cancer, was 2.37 ± 0.68. Compared to the control, 1.0% or 2.0% calcium treatment significantly decreased the number of total ACFs and ACs in a concentration-dependent manner. The decrease in ACFs or ACs after calcium treatment was associated with decreases in cell proliferation and β-catenin expression and an increase in apoptosis in colonic mucosal cells. These results suggest that calcium may exert a protective effect against colon cancer by inhibiting the development of ACFs/ACs in ICR mice.

Selenium (Se) is known to prevent from several cancers, while iron (Fe) is known to be associated with high risk of cancers. The role of Se on colon carcinogenesis was investigated in an animal model induced by azoxymethane (AOM) and dextran sodium sulfate (DSS) in low Fe mice. Six-week old ICR mice fed on a low Fe diet (4.5 ppm Fe; generally 10 times lower than normal Fe) with three different Se (0.02, 0.1 or 0.5 ppm) levels for 24weeks. The animals received weekly three (0~2nd weeks) i.p. injections of AOM (10 mg/kg B.W), followed by 2%DSS with drinking water for 1 week to induce the colon cancer. There were five experimental groups including vehicle,positive control (normal Fe level, AOM/DSS), Low Fe (LFe) + AOM/DSS+Low Se (LSe), LFe + AOM/DSS + medium Se (MSe) and LFe + AOM/DSS + high Se (HSe) groups. HSe group showed a 66.7% colonic tumor incidence, MSe group showed a 69.2% tumor incidence, and LSe group showed a 80.0% tumor incidence. The tumor incidence was negatively associated with Se levels of diets. Tumor multiplicity in Hse group was significantly low compared to the other groups (p < 0.05). With increasing Se levels of diets, the primary anti-proliferating cell nuclear antigen (PCNA)-positive cells were decreased and apoptotic bodies were increased in a dose-dependent manner. Sedependent glutathione peroxidase activity and its protein level were dependent on the levels of Se of diets. Malondialdehyde level in liver was lowest in Hse group among experimental groups. These findings indicate that dietary Se is chemopreventive for colon cancer by increasing antioxidant activity and decreasing cell proliferation in Fe-deficient mice.

본 연구에서는 azoxymethane (AOM)과 dextran sodium sulfate (DSS)로 유도된 대장 발암과정에 대한 셀레늄의 방어 효과를 조사하였다. 셀레늄 결핍(0.02 ppm Se), 정상(0.1 ppm Se), 과다(0.5 ppm Se)사료를 12주간 식이로 급여하여 혈액검사와 대장암 발생의 초기단계인 aberrant crypt foci (ACF)수를 측정했으며, 암 발생율을 조사하였다. ICP-AES 를 사용하여 간의 셀레늄 농도를 측정하였으며, 또한 셀레늄포함 항산화효소인 glutathione peroxidase (GPx) 활성을 알아보았다. 또한 TUNEL assay와 PCNA, β-catenin에 대한 면역조직 염색을 수행하였다. ACF 수 및 종양 발생률에 있어서, 셀레늄과다사료를 급여한 군이 정상셀레늄사료를 급여한 군보다 낮았으며, 셀레늄결핍사료를 급여한 군은 오히려 ACF 수 및 종양 발생률이 높았다. GPx 활성은 셀레늄의 섭취가 과다한 군에서 높게 나타났으며, 이 때, TUNEL 에서 apoptotic positive cell이 증가하는 것을 확인했다. 또 한 셀레늄의 섭취가 과다한 군에서 PCNA와 β-catenin의 발현이 감소됨을 볼 수 있었다. 본 마우스 모델실험에서 셀레늄은 여러 기전에 의해 대장암 발생을 억제할 수 있을 것으로 사료된다.