1. Top of page
  2. Abstract

The usefulness of mucin-depleted foci (MDF), which have recently been proposed as a new preneoplastic biomarker in rat colon carcinogenesis, was histologically investigated in rat colonic tissues treated with 1,2-dimethylhydrazine dihydrochloride (DMH). The relationship among aberrant crypt foci (ACF), MDF and β-catenin accumulated crypts (BCAC) was examined by comparing the corresponding computer-captured images. Twelve male F344 rats were given DMH s.c. at a dose of 40 mg/kg body weight, once a week for 2 weeks, and randomly divided into two groups. Rats in group 1 were given normal drinking water, while those in group 2 were given drinking water containing indomethacin (IND) at 16 ppm for 6 weeks. All animals were sacrificed 8 weeks after the first DMH treatment. The resected colons were fixed in 10% formalin, and stained with Alcian blue for observation of ACF and MDF. Histological and immunohistochemical analysis revealed that the numbers of ACF, MDF and overlapping lesions in group 2 (treated with IND) were significantly decreased, compared with those in group 1. The number of BCAC in group 2 was also significantly lower than that in group 1. The reduction (61.5%) of MDF by IND was much greater than that (29.3%) of ACF. Analyses of the computer-captured images indicated that MDF had more frequent dysplastic changes and overexpression of β-catenin than did ACF. MDF having over 4 crypts or MDF with the appearance of ACF corresponded well to BCAC. These results suggest that MDF may be useful as an early biomarker in colon carcinogenesis.


Alcian blue pH 2.5


aberrant crypt foci


β-catenin accumulated crypts


dysplastic foci


mucin-depleted foci


1,2-dimethylhydrazine dihydrochloride


hematoxylin and eosin


high-iron di-amine Alcian blue




normal-like crypts

  • 1
    Bird RP. Observation and quantification of aberrant crypts in the marine colon treated with a colon carcinogen: preliminary findings. Cancer Lett 1987; 37: 14751.
  • 2
    Corpet DE, Tache S. Most effective colon cancer chemopreventive agents in rats: a systematic review of aberrant crypt foci and tumor data, ranked by potency. Nutr Cancer 2002; 43: 121.
  • 3
    Magnuson BA, Carr I, Bird RP. Ability of aberrant crypt foci characteristics to predict colonic tumor incidence in rats fed cholic acid. Cancer Res 1993; 53: 4499504.
  • 4
    Zheng Y, Kramer PM, Lubet RA, Steele VE, Kelloff GJ, Pereira MA. Effect of retinoids on AOM-induced colon cancer in rats: modulation of cell proliferation, apoptosis and aberrant crypt foci. Carcinogenesis 1999; 20: 25560.
  • 5
    Caderni G, Femia AP, Giannini A, Favuzza A, Luceri C, Salvadori M, Dolara P. Identification of mucin-depleted foci in the unsectioned colon of azoxymethane-treated rats: correlation with Carcinogenesis. Cancer Res 2003; 63: 238892.
  • 6
    Yamada Y, Yoshimi N, Hirose Y, Kawabata K, Matsunaga K, Shimizu M, Hara A, Mori H. Frequent beta-catenin gene mutations and accumulations of the protein in the putative preneoplastic lesions lacking macroscopic aberrant crypt foci appearance, in rat colon Carcinogenesis. Cancer Res 2000; 60: 33237.
  • 7
    Yamada Y, Yoshimi N, Hirose Y, Matsunaga K, Katayama M, Sakata K, Shimizu M, Kuno T, Mori H. Sequential analysis of morphological and biological properties of beta-catenin-accumulated crypts, provable premalignant lesions independent of aberrant crypt foci in rat colon Carcinogenesis. Cancer Res 2001; 61: 18748.
  • 8
    Takahashi M, Fukuda K, Sugimura T, Wakabayashi K. Beta-catenin is frequently mutated and demonstrates altered cellular location in azoxymethane-induced rat colon tumors. Cancer Res 1998; 58: 426.
  • 9
    Suzui M, Ushijima T, Dashwood RH, Yoshimi N, Sugimura T, Mori H, Nagao M. Frequent mutations of the rat beta-catenin gene in colon cancers induced by methylazoxymethanol acetate plus 1-hydroxyanthraquinone. Mol Carcinog 1999; 24: 2327.
  • 10
    Suzui M, Sugie S, Mori H, Okuno M, Tanaka T, Moriwaki H. Different mutation status of the beta-catenin gene in carcinogen-induced colon, brain, and oral tumors in rats. Mol Carcinog 2001; 32: 20612.
  • 11
    Dashwood RH, Suzui M, Nakagama H, Sugimura T, Nagao M. High frequency of beta-catenin (ctnnbl) mutations in the colon tumors induced by two heterocyclic amines in the F344 rat. Cancer Res 1998; 58: 11279.
  • 12
    Harada N, Tamai Y, Ishikawa T, Sauer B, Takaku K, Oshima M, Taketo MM. Intestinal polyposis in mice with a dominant stable mutation of the beta-catenin gene. EMBO J 1999; 18: 593142.
  • 13
    Femia AP, Dolara P, Caderni G. Mucin-depleted foci (MDF) in the colon of rats treated with azoxymethane (AOM) are useful biomarkers for colon Carcinogenesis. Carcinogenesis 2004; 25: 27781.
  • 14
    Morioka T, Suzui M, Nabandith V, Inamine M, Aniya Y, Nakayama T, Ichiba T, Mori H, Yoshimi N. The modifying effect of Peucedanum japonicum, a herb in the Ryukyu Islands, on azoxymethane-induced colon preneoplastic lesions in male F344 rats. Cancer Lett 2004; 205: 13341.
  • 15
    Kawabata K, Tanaka T, Murakami T, Okada T, Murai H, Yamamoto T, Hara A, Shimizu M, Yamada Y, Matsunaga K, Kuno T, Yoshimi N, Sugie S, Mori H. Dietary prevention of azoxymethane-induced colon Carcinogenesis with rice-germ in F344 rats. Carcinogenesis 1999; 20: 210915.
  • 16
    Yoshimi N, Shimizu M, Matsunaga K, Yamada Y, Fujii K, Kara A Mori, H. Chemopreventive effect of N-(2-cyclohexyloxy-4-nitrophenyl)methane sul-fonamide (NS-398), a selective cyclooxygenase-2 inhibitor, in rat colon Carcinogenesis induced by azoxymethane. Jpn J Cancer Res 1999; 90: 40612.
  • 17
    Yamada Y, Yoshimi N, Hirose Y, Hara A, Shimizu M, Kuno T, Katayama M, Qiao Z, Mori H. Suppression of occurrence and advancement of beta-catenin-accumulated crypts, possible premalignant lesions of colon cancer, by selective cyclooxygenase-2 inhibitor, celecoxib. Jpn J Cancer Res 2001; 92: 61723.
  • 18
    Takahashi M, Mutoh M, Kawamori T, Sugimura T, Wakabayashi K. Altered expression of β-catenin, inducible nitric oxide synthase and cyclooxygenase-2 in azothymethane-induced rat colon Carcinogenesis. Carcinogenesis 2000; 21: 131927.
  • 19
    Ohchiai M, Ushigome M, Fujiwara K, Ubegai T, Kawamori T, Sugimura T, Nagao N, Nakagama H. Characterization of dysplastic aberrant crypt foci in the rat colon induced by 2-amino-l-methyl-6-phenylimidazo[4,5-b]pyridine. Am J Pathol 2003; 163: 160714.
  • 20
    Takahashi M, Wakabayashi K. Gene mutations and altered gene expression in azoxymethane-induced colon Carcinogenesis in rodents. Cancer Sci 2004; 95: 47580.
  • 21
    Hirose Y, Kuno T, Yamada Y, Sakata K, Katayama M, Yoshida K, Qiao Z, Hata K, Yoshimi N, Mori H. Azoxymethane-induced beta-catenin-accumu-lated crypts in colonic mucosa of rodents as an intermediate biomarker for colon carcinogenesis. Carcinogenesis 2003; 24: 10711.
  • 22
    Yamada Y, Mori H. Pre-cancerous lesions for colorectal cancers in rodents: a new concept. Carcinogenesis 2003; 24: 10159.
  • 23
    Gendler SJ, Spicer AP. Epithelial mucin genes. Annu Rev Physiol 1995; 57: 60734.
  • 24
    Kim YS, Gum J Jr, Brockhausen I. Mucin glycoproteins in neoplasia. Glyco-conj J 1996; 13: 693707.
  • 25
    Zusman I, Zimber A, Nyska A. Role of morphological methods in the analysis of chemically induced colon cancer in rats. Acta Anat (Basel) 1991; 142: 3516.
  • 26
    Dawson PA, Patel J, Filipe MI. Variations in sialomucins in the mucosa of the large intestine in malignancy: a quantimet and statistical analysis. His-tochem J 1978; 10: 55972.
  • 27
    Filipe MI. Value of histochemical reactions for mucosubstances in the diagnosis of certain pathological conditions of the colon and rectum. Gut 1969; 10: 57786.
  • 28
    Matsushita Y, Yamamoto N, Shirahama H, Tanaka S, Yonezawa S, Yamori T, Irimura T, Sato E. Expression of sulfomucins in normal mucosae, colorectal adenocarcinomas, and metastases. Jpn J Cancer Res 1995; 86: 10607.
  • 29
    Sandforth F, Heimpel S, Balzer T, Gutschmidt S, Riecken EO. Characterization of stereomicroscopically identified preneoplastic lesions during dimeth-ylhydrazine-induced colonic carcinogenesis. Eur J Clin Invest 1988; 18: 65562.
  • 30
    Filipe MI. Mucous secretion in rat colonic mucosa during carcinogenesis induced by dimethylhydrazine. A morphological and histochemical study. Br J Cancer 1975; 32: 6077.