Dietary Selenium Supplement Prevents Colon Carcinogenesis Induced by Azoxymethane and Dextran Sodium Sulfate in ICR Mice

Jun-hyeong Kim; Jin-Joo Hue; Bong Su Kang; Hyunji Park; Sang Yoon Nam; Young Won Yun; Jong Soo Kim; Jae-Hwang Jeong; Beom Jun Lee

doi:10.5625/lar.2010.26.3.293

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Kim, Hue, Kang, Park, Nam, Yun, Kim, Jeong, and Lee: Dietary Selenium Supplement Prevents Colon Carcinogenesis Induced by Azoxymethane and Dextran Sodium Sulfate in ICR Mice

Original Article

Lab. Anim. Res. 2010;26(3):293-300.

Published online: 17 September 2010

DOI: https://doi.org/10.5625/lar.2010.26.3.293

Dietary Selenium Supplement Prevents Colon Carcinogenesis Induced by Azoxymethane and Dextran Sodium Sulfate in ICR Mice

Jun-hyeong Kim¹, Jin-Joo Hue¹, Bong Su Kang¹, Hyunji Park¹, Sang Yoon Nam¹, Young Won Yun¹, Jong Soo Kim¹, Jae-Hwang Jeong², Beom Jun Lee^1,^∗

¹Department of Veterinary Medicine, College of Veterinary Medicine and Research Institute of Veterinary Medicine, Chungbuk National University, Cheongju, Korea

²Department of Biotechnology and Biomedicine, Chungbuk Province College, Okcheon, Korea

^∗Corresponding author: Beom Jun Lee, Department of Veterinary Medicine, College of Veterinary Medicine, Chungbuk National University, 410 Seongbongro (Gaeshin-dong), Cheongju, Chungbuk 361-763, Korea Tel: +82-43-261-3357 Fax: +82-43-271-3246 E-mail: beomjun@cbu.ac.kr

Received 31 August 2010 Revised 17 September 2010 Accepted 17 September 2010

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

The role of selenium (Se) in modulating colon carcinogenesis induced by azoxymethane (AOM) followed by dextran sodium sulfate (DSS) was investigated in mice. Five-week old ICR mice were fed on diets containing different concentrations (0.02, 0.1 or 0.5 ppm) of Se for 24 weeks. Animals received three (0-2nd weeks) intraperitoneal injections of AOM (10 mg/kg body weight), followed by 2% DSS with drinking water for additional 1 week. There were 4 experimental groups including vehicle control group, positive control group given AOM/DSS with AIN-93G normal diet containing 0.1% Se (NSe), a low (0.02 ppm)-Se diet group (LSe) and a high (0.5 ppm)-Se diet group (HSe). Hematology was analyzed with a blood cell differential counter. Liver Se was analyzed by inductively coupled plasma-mass spectroscopy. Cell proliferation and apoptosis were determined by using proliferating cell nuclear antigen (PCNA) for proliferative activity and apoptotic index by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL), respectively. HSe group showed a low incidence of colonic tumor (64.7%), compared with the NSe positive control (75%) and LSe (77.8%) groups. In contrast, HSe group exhibited lower rate of PCNA-positive cells (39.3±6.9%) than positive control (64.3±0.3%) and LSe (57.3±2.9%) groups. In addition, apoptotic index of HSe group was higher than those of positive control and LSe groups. These results indicate that Se is a chemopreventive agent for colon carcinogenesis induced by AOM+DSS in male ICR mice.

Keywords: Azoxymethane (AOM), dextran sodium sulfate (DSS), colon cancer, selenium, cell proliferation, apoptosis

REFERENCES

Abdulah R.., Miyazaki K.., Nakazawa M.., Koyama H.2005. ).Chemical forms of selenium for cancer prevention. J. Trace Elem. Med. Biol. 19(2-3):141–150.

Beno I.., Klvanova J.., Magalova T.., Brtkova A.2000. Blood levels of natural antioxidants in gastric and colorectal precancerous lesions and cancers in Slovakia. Neoplasma. 47(1):37–40.

Bravo R.., Frank R.., Blundell P.A.., Macdonald-Bravo H.1987. Cyclin/PCNA is the auxiliary protein of DNA polymerase-delta. Nature. 326(6112):515–517.

Choi Y.S.., Hesketh J.E.2006. Nutritional Biochemistry of selenium. J. Kor. Soc. Food Sci. Nutr. 35(5):661–670.

Clark L.C.., Combs G.F. Jr.., Turnbull B.W.., Slate E.H.., Chalker D.K.., Chow J.., Davis L.S.., Glover R.A.., Graham G.F.., Gross E.G.., Krongrad A.., Lesher J.L. Jr.., Park H.K.., Sanders B.B. Jr.., Smith C.L.., Taylor J.R.1996. Effects of selenium supplementation for cancer prevention in patients with carcinoma of the skin. A randomized controlled trial. Nutritional Prevention of Cancer Study Group. JAMA. 276(24):1957–1963.

Combs G.F. Jr.2005. Current evidence and research needs to support a health claim for selenium and cancer prevention. J. Nutr. 135(2):343–347.

Combs G.F. Jr.., Combs S.B.1984. The nutritional biochemistry of selenium. Annu. Rev. Nutr. 4:257–280.

Davis C.D.., Johnson W.T.2002. Dietary copper affects azoxymethane-induced intestinal tumor formation and protein kinase C isozyme protein and mRNA expression in colon of rats. J. Nutr. 132(5):1018–1025.

Davis R. L.., Spallholz J. E.1996. Inhibition of selenite-catalyzed superoxide generation and formation of elemental selenium (Se^o) by copper, zinc, and aurintricarboxylic acid (ATA). Biochem. Pharmacol. 51(8):1015–1020.

Finley J.W.1999. The retention and distribution by healthy young men of stable isotopes of selenium consumed as selenite, selenate or hydroponically-grown broccoli are dependent on the isotopic form. J. Nutr. 129(4):865–871.

Ip C.., Dong Y.., Ganther H.E.2002. ).New concepts in selenium chemoprevention. Cancer Metastasis Rev. 21(3-4):281–289.

Ip C.., Hayes C.., Budnick R.M.., Ganther H.E.1991. Chemical form of selenium, critical metabolites, and cancer prevention. Cancer Res. 51(2):595–600.

Jemal A.., Murray T.., Ward E.., Samuels A.., Tiwari R.C.., Ghafoor A.., Feuer E.J.., Thun M.J.2005. Cancer statistics, 2005. CA Cancer J. Clin. 55(1):10–30.

Jiang C.., Jiang W.., Ip C.., Ganther H.., Lu J.1999. selenium -induced inhibition of angiogenesis in mammary cancer at chemopreventive levels of intake. Mol. Carcinog. 26(4):213–225.

Korea National Statistical Office. 2007. Death and Cause of Death Statistics 2006. KNSO, Daejeon.

Milde D.., Novak O.., Stu ka V.., Vyslou il K.., Macha ek J.2001. Serum levels of selenium, manganese, copper, and iron in colorectal cancer patients. Biol. Trace Elem. Res. 79(2):107–114.

National Institute of Occupational Safety and Health (NIOSH). 1990. Registry of Toxic Effects of Chemical Substances (RTECS). Canadian Centre for Occupational Health and Safety, Hamilton, Ontario.

Okayasu I.., Hatakeyama S.., Yamada M.., Ohkusa T.., Inagaki Y.., Nakaya R.1990. A novel method in the induction of reliable experimental acute and chronic ulcerative colitis in mice. Gastroenterology. 98(3):694–702.

Patrick L.2004. selenium biochemistry and cancer: a review of the literature. Altern. Med. Rev. 9(3):239–258.

Tanaka T.., de Azevedo M.B.., Duran N.., Alderete J.B.., Epifano F.., Genovese S.., Tanaka M.., Curini M.2010. Colorectal cancer chemoprevention by 2 beta-cyclodextrin inclusion compounds of auraptene and 4'-geranyloxyferulic acid. Int. J. Cancer. 126(4):830–840.

Tanaka T.., Kohno H.., Suzuki R.., Yamada Y.., Sugie S.., Mori H.2003. A novel inflammation-related mouse colon carcinogenesis model induced by azoxymethane and dextran sodium sulfate. Cancer Sci. 94(11):965–973.

Pawlowicz Z.., Zachara B.A.., Trafikowska U.., Nowicki A.1993. ).Low levels of selenium and activity of glutathione peroxidase in blood of patients with gastrointestinal neoplasms. Pol. Tyg. Lek. 48(25-26):554–556.

Rayman M.P.2005. selenium in cancer prevention: a review of the evidence and mechanism of action. Proc. Nutr. Soc. 64(4):527–542.

Rosenberg D.W.., Giardina C.., Tanaka T.2009. Mouse models for the study of colon carcinogenesis. Carcinogenesis. 30(2):183–196.

Rudolf E.., Kralova V.., Cervinka M.2008. selenium and colon cancer–from chemoprevention to new treatment modality. Anticancer Agents Med. Chem. 8(6):598–602.

Schrauzer G.N.2000. ).Anticarcinogenic effects of selenium. Cell. Mol. Life Sci. 57(13-14):1864–1873.

Sohn O.S.., Fiala E.S.., Requeijo S.P.., Weisburger J.H.., Gonzalez F.J.2001. Differential effects of CYP2E1 status on the metabolic activation of the colon carcinogens azoxymethane and methylazoxymethanol. Cancer Res. 61(23):8435–8440.

Figure 1.

Experimental design for colon carcinogenesis in mice. AOM, azoxymethane; DSS, dextran sodium sulfate.

Figure 2.

Change in the body weights in mice treated with AOM/DSS and selenium (Se). The body weights of all AOM/DSS treatment groups significantly decreased compared with the body weight of vehicle control group during 2–4th weeks of treatment (P<0.05). AOM: azoxymethane, DSS: dextran sodium sulfate, NSe: normal selenium (0.1 ppm) diet, LSe: low selenium (0.02 ppm) diet, HSe: high selenium (0.5 ppm) diet.

Figure 3.

Hepatic selenium levels in mice following feeding of different concentrations of dietary selenium. Selenium concentration was determined using an inductively coupled plasma-mass spectroscopy. AOM: azoxymethane, DSS: dextran sodium sulfate, NSe: normal selenium (0.1 ppm) diet, LSe: low selenium (0.02 ppm) diet, HSe: high selenium (0.5 ppm) diet.

Figure 4.

Effect of selenium on tumor incidence and survival rate of mice treated with azoxymethane/dextran sodium sulfate (AOM/DSS). NSe: normal selenium (0.1 ppm) diet, LSe: low selenium (0.02 ppm) diet, HSe: high selenium (0.5 ppm) diet.

Figure 5.

Immunohistochemistry on proliferating cell nuclear antigen (PCNA) in the colon of mice treated with azoxymethane/dextran sodium sulfate (AOM/DSS) and selenium (Se). The PCNA-positive cells were greatly increased by treatment with AOM/DSS, which were markedly reduced by coadministration of a high concentration (0.5 ppm) of selenium (Se). (A) Vehicle control. (B) AOM/DSS+normal Se (0.1 ppm) diet. (C) AOM/DSS+low Se (0.02 ppm) diet. (D) AOM/DSS+ high Se (0.5 ppm) diet. ×100.

Figure 6.

TUNEL assay for apoptotic nuclei in distal colon sections of mice treated with azoxymethane/dextran sodium sulfate (AOM/DSS) and selenium (Se). The TUNEL-positive cells were increased by treatment with AOM/DSS, which were further enhanced by co-administration of Se in a concentration-dependent manner. (A) Vehicle control. (B) AOM/DSS+normal Se (0.1 ppm) diet. (C) AOM/DSS+low Se (0.02 ppm) diet. (D) AOM/DSS+high Se (0.5 ppm) diet. ×100.

Table 1.

Composition of the diets at different selenium status

Ingredients (g/kg diet)	NSe	LSe	HSe
Ingredients (g/kg diet)	g/kg	g/kg	g/kg
Casein	200	200	200
L-Cystine	3	3	3
Sucrose	88.9	88.98	88.5
Cornstarch	397.486	397.486	397.486
Dyetrose	132	132	132
Soybean oil t-Butylhydroquinone	70 0.014	70 0.014	70 0.014
Microcrystalline cellulose	50	50	50
Mineral mix (Rx grade, no Fe, Se)	37	37	37
Ferric citrate 5 mg/g as Fe	9	9	9
Sodium selenate 1 mg/g as Se	0.1	0.02	0.5
Vitamin mix	10	10	10
Choline bitartrate	2.5	2.5	2.5
Total	1000	1000	1000

Fe: iron, Se: selenium.

Table 2.

Differential blood cell counts in ICR mice fed different selenium diets

Parameters	Vehicle	AOM/DSS
Parameters	Vehicle	NSe	LSe	HSe
WBC (10³/µL)	4.61±1.85	2.61±0.61	2.13±0.55	2.32±0.20
RBC (10⁶/µL)	8.72±0.40	07.11±1.66^∗	7.72±0.23	7.99±0.57
Hemoglobin (g/dL)	13.40±0.600	11.60±1.13^∗	12.23±0.250	12.20±0.600
Hematocrit (%)	44.05±2.800	38.20±4.67^∗	40.47±1.760	40.97±1.440
MCV (fl)	50.53±0.940	54.50±6.220	52.43±2.700	51.40±2.170
MCV (fl) MCH (pg)	50.53±0.940 15.40±0.550	54.50±6.220 16.65±2.330	52.43±2.700 15.87±0.400	51.40±2.170 15.33±0.350
MCHC (g/dL)	30.53±1.230	30.81±0.680	30.30±0.890	29.87±0.760

WBC: white blood cells, RBC: red blood cells, MCV: mean corpuscular volume, MCH: mean corpuscular hemoglobin, MCHC: mean corpuscular hemoglobin concentration, AOM: azoxymethane, DSS: dextran sodium sulfate, NSe: normal selenium (0.1 ppm) diet, LSe: low selenium (0.02 ppm) diet, HSe: high selenium (0.5 ppm) diet.

^∗ Significantly different from vehicle control (P<0.05).

Table 3.

Effect of selenium on cell proliferative and apoptotic indices in AOM/DSS- induced colon carcinogenesis

Groups	Selenium (ppm)	Proliferative index (%)	Apoptotic index (%)
Vehicle	0.1	20.33±2.91	1.33±0.67
AOM/DSS+NSe	0.1	∗64.33±0.33∗	∗8.33±0.67∗
AOM/DSS+LSe	0.02	57.33±2.85	∗4.50±0.41^†
AOM/DSS+HSe	0.5	∗39.33±6.94^†	11.25±1.28^†.

AOM: azoxymethane, DSS: dextran sodium sulfate, NSe: normal selenium (0.1 ppm) diet, LSe: low selenium (0.02 ppm) diet, HSe: high selenium (0.5 ppm) diet.

^† Significantly different from AOM/DSS+NSe (P<0.05).

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