Colorectal carcinogenesis is a complex multistep process that includes changes in histomorphological appearance of the colonic mucosa and changes at molecular level. Aberrant crypt foci (ACF) was first described by Bird in 1987 on examination of methylene-blue-stained colonic mucosa of azoxymethane-treated mice under light microscopy. Since then ACF was considered as the earliest preneoplastic change that can be seen in the colonic mucosa. The aim of this study was to look at the histomorphology and distribution of ACF in colorectal carcinoma. 50 formalin-fixed archival colectomy specimens for colorectal carcinoma were examined under light microscopy after staining with 0.2% methylene blue. ACF was identified by larger and darker crypts with thickened epithelium, and often elevated from adjacent normal mucosa. ACF was found in 41 of 50 colectomy specimens examined. There were 328 ACF consisting of 36 (11.0%) ACF without hyperplasia or dysplasia, 263 (80.2%) ACF with hyperplasia and 29 (8.8%) ACF with dysplasia. Of these 29 ACF with dysplasia, 25 showed low grade dysplasia and four high grade dysplasia. The density of ACF was higher in the left colon, those older than 65 years of age and among males but these findings were statistically not significant. The crypt multiplicity of hyperplastic ACF (30.149, SD 28.395) was larger than dysplastic ACF (20.613, SD 40.128). The spectrum of histological changes observed probably represent the evolution of ACF in colorectal carcinogenesis.
Aberrant Crypt Foci/*pathology ; Adenocarcinoma/*pathology ; Colorectal Neoplasms/*pathology

BACKGROUND/AIMS: The purpose of this study was to investigate the malignant potential of aberrant crypt foci (ACF) by measuring the multiplicity of crypts and lectin expression in the early and late stages of 1,2-dimethylhydrazine (DMH)-induced colon carcinogenesis. METHODS: Six-week-old Wistar rats were injected subcutaneously with DMH for 27 weeks. We classified ACF according to the number of crypts per ACF as a few crypts (< or =3 crypts, FC ACF) or numerous crypts (> or =4 crypts, NC ACF). Immunohistochemistry was used to evaluate lectin expression. RESULTS: In the early stage, FC ACF (590/1,902, 31.0%) occurred more frequently than NC ACF (35/449, 7.8%); whereas in the late stage, NC ACF (176/449, 39.2%) occurred more frequently than FC ACF (324/1,902, 17.0%). The number of ACF peaked at 15 to 20 weeks. The ratio of NC/FC ACF increased gradually during carcinogenesis. The expression of both UEA1 and PNA was higher in NC ACF than FC ACF. Lectin expression increased in the late stage compared with the early stage. CONCLUSIONS: The expression of lectin was higher in NC ACF and ACF in the late stage. Therefore, ACF with higher multiplicities in the late stage may have more malignant potential in DMH-induced colon carcinogenesis.
1,2-Dimethylhydrazine ; Aberrant Crypt Foci ; Animals ; Colon ; Dimenhydrinate ; Immunohistochemistry ; Peanut Agglutinin ; Rats ; Rats, Wistar

BACKGROUND/AIMS: Aberrant crypt foci (ACF) are early microscopic lesions of the colonic mucosa, which can be detected by magnified chromoendoscopy. Herein, we have investigated whether ACF identified in different clinical groups can be differentiated based on their characteristics. METHODS: Macroscopically unremarkable mucosal flaps were collected from 270 fresh colectomies and divided into 3 clinical groups: colorectal carcinoma (group A), disease controls having known pre-neoplastic potential (group Bc), and disease controls without risk of carcinoma development (group Bn). Topographic and histologic analysis, immunohistochemistry, and molecular studies (high-resolution melt curve analysis, real-time polymerase chain reaction, and Sanger sequencing) were conducted for certain neoplasia-associated markers. RESULTS: ACF were seen in 107 cases, out of which 72 were left colonic ACF and 35 right colonic ACF (67.2% vs. 32.7%, P=0.02). The overall density of left colonic ACF was 0.97/cm, which was greater than the right colonic ACF density of 0.81/cm. Hypercrinia was present in 41 out of 72 left colonic ACF and in 14 out of 35 right colonic ACF (P=0.01). Immunohistochemical expression of p53 was also greater in left colonic ACF than in right colonic ACF (60.5% vs. 38.2%, P=0.03). However, ACF identified among the 3 clinical groups did not show any distinguishing topographic, histological, or genetic changes. CONCLUSIONS: Left colonic ACF appear to be high-risk based on their morphological and prototypic tumor marker signature. ACF identified in different clinical groups do not show significant genotypic or topographic differences. Further detailed genetic studies are required to elucidate them further.
Aberrant Crypt Foci ; Colectomy ; Colon ; Colorectal Neoplasms ; Humans ; Immunohistochemistry ; Mucous Membrane ; Real-Time Polymerase Chain Reaction

Aberrant crypt foci (ACF) are grossly unidentifiable lesions of the colon and visible only with low-power microscopic examinations after methylene blue stain. To establish the role of ACF in colorectal carcinogenesis, we evaluated the distribution, frequency, histopathological classification, and patterns of mucin secretion of ACF in the colon. A total of 142 aberrant crypt foci were found in 41 colectomy specimen for adenocarcinoma (36 cases) and benign diseases of colon (5 cases). Ten of 142 ACFs were in the ascending and transverse colon, 39 in the descending and sigmoid colon, and 93 in the rectum. The mean number of ACFs in the rectum (0.13 0.11/cm2) was higher than in the ascending and transverse colons (0.019 0.018/ cm2) and descending and sigmoid colon (0.10 0.14/cm2). ACFs were found only in cancer patients. One hundred and twenty ACFs among 142 ACFs identified by topology, were identified on histological examination. We classified ACFs into simple (48.3%), hyperplastic (42.5%), and dysplastic (9.2%) types. All ACFs were infiltrated by the lymphocytes in the stroma and 18 of these accompanied the lymphoid follicles. ACFs have variable histopathologic features and mucin profiles. Some variants of ACFs are at the early stage of the spectrum between benign and malignant.
Aberrant Crypt Foci* ; Adenocarcinoma ; Carcinogenesis ; Classification* ; Colectomy ; Colon ; Colon, Sigmoid ; Colon, Transverse ; Humans ; Lymphocytes ; Methylene Blue ; Mucins* ; Rectum

We investigated the combinatorial effects of different doses of dietary soy isoflavones (SI) and fructooligosaccharide (FOS) in a rat model of colon cancer. We hypothesized that increased bioavailability of SI metabolites due to dietary FOS may increase production of bioactive equol and affect colon carcinogenesis in a dose-dependent manner. Sprague-Dawley male rats were injected with 12-dimethylhydrazine (DMH) and were provided experimental diets that contained 0, 10, 50, 150, or 500 mg SI per kg of diet and 6% FOS for 12 weeks. The number of aberrant crypt foci (ACF) and the expression of cyclooxygenase-2 (COX-2) in colonic tissues were significantly decreased in the 6% FOS-fed groups compared to the control group. Gut transit time and fecal pH were significantly lower, and fecal concentrations of bifidobacteria were increased with 6% FOS. However, dietary SI supplementation in combination with 6% dietary FOS did not affect ACF formation or COX-2 expression. Plasma equol concentrations were dose-dependently increased by supplementation of SI up to 500 mg/kg of diet. In conclusion, SI supplementation up to 500 mg/kg of diet appeared to have no additive beneficial effects in rats with chemically-induced colon cancer that were fed 6% FOS, although plasma equol was dose-dependently increased.
Aberrant Crypt Foci ; Animals ; Biological Availability ; Colon ; Colonic Neoplasms ; Cyclooxygenase 2 ; Diet ; Equol ; Humans ; Hydrogen-Ion Concentration ; Isoflavones ; Male ; Oligosaccharides ; Plasma ; Rats

PURPOSE: DNA-PKcs is one of the DNA repair genes. It was recently found that hyperplasia and dysplasia of the intestinal mucosa and the production of aberrant crypt foci were developed in DNA-PKcs-null mice, and this suggests a suppressive role for DNA-PKcs in tumorigenesis. MATERIALS AND METHODS: To investigate the possible relationship between the clinico-pathologic characteristics and the survival of gastric cancer patients, the expression status of DNA-PKcs was determined in 279 consecutive gastric cancers. Immunohistochemical analysis was performed to evaluate the expression levels of DNA-PKcs protein by using the tissue array method. RESULTS: Out of 279 consecutive gastric cancers, 63 cases (22.6%) showed the loss of DNA-PKcs expression. The loss of DNA-PKcs expression was significantly associated with advanced cancer (p <0.001), lymphatic invasion (p=0.001), lymph node metastasis (p=0.009), and advanced pTNM stage (p=0.009). Univariate survival analysis revealed that patients with the loss of DNA-PKcs expression had significantly poorer survival than those patients with intact DNA-PKcs expression (p=0.004). Moreover, the loss of DNA-PKcs expression was identified to correlate with a lower survival in the subgroup of stage I gastric cancer patients (p=0.037). CONCLUSION: The loss of DNA-PKcs expression was found in 23% of human gastric cancers and this was identified to significantly correlate with poor patient survival, especially for stage I gastric cancer patients.
Aberrant Crypt Foci ; Animals ; Carcinogenesis ; Catalytic Domain* ; DNA Repair ; DNA-Activated Protein Kinase* ; Humans ; Hyperplasia ; Immunohistochemistry ; Intestinal Mucosa ; Lymph Nodes ; Mice ; Neoplasm Metastasis ; Stomach Neoplasms* ; Survival Analysis

PURPOSE: This study was performed in order to determine the histologic features, incidence & frequency of the Aberrant Crypt Foci (ACFs) including mucosal abnormalities arising in the sporadic colonic cancer. MATERIALS AND METHODS: We investigated the proximal and distal colonic mucosa surrounding the tumor in 22 cases (right colon 7 cases and left colon 15 cases) of resected colonic adenocarcinoma specimen. The methylene blue- stained colonic mucosa was examined in en face preparations and rolled totally. The rolled colonic mucosa was embedded in paraffin and examined by using 4micrometer thick serial sections. RESULTS: We found one hundred twenty two ACFs. The 97 foci (78.7%) were simple hypertrophic foci (SH), composed of more elongated and larger crypts than normal with apical branching associated with goblet cell hyperplasia. The 17 foci (13.9%) were hyperplastic foci (HP) resembling hyperplastic polyp, and 7 (5.7%) were adeno matous foci (AD) while 2 (1.6%) were adenomatous foci with dysplasia (Dys). The mean number of ACFs/cm of the examined mucosa were 0.18+/-0.21 and were higher in the left colon than in the right colon (0.22+/-0.24 vs. 0.10+/-0.10). Immunohistochemical stains for p53 and Ki-67 in these foci revealed strong and upper cryptal staining patterns in AD and Dys of ACFs, like that of neoplasia or preneoplastic condition. However, the staining intensities in SH and HP of ACFs were equal to or lower than that of normal crypts. CONCLUSION: These results suggest that grossly defined ACFs include reactive process and the majority of ACFs are induced by simple reactive alteration without preneoplastic potential; and two types of ACFs (AD and Dys) are more likely to be direct precursors of colon tumors than SH or HP.
Aberrant Crypt Foci* ; Adenocarcinoma* ; Colon ; Colonic Neoplasms ; Colorectal Neoplasms ; Coloring Agents ; Goblet Cells ; Hyperplasia ; Incidence ; Mucous Membrane* ; Paraffin ; Polyps ; Precancerous Conditions

Colorectal cancer (CRC) is one of the leading causes of cancer death in western countries or in the developed countries. Zinc intake has been associated with decreased risk of CRC. We investigated the effect of zinc on the formation of colonic aberrant crypt foci (ACF) induced by azoxymethane followed by dextran sodium sulfate in mice. Five-week old ICR mice were fed with the different zinc levels (0.01, 0.1, 1 ppm) for 12 weeks. The numbers of ACF were measured in the colonic mucosa. The ACF number of HZn group was significantly low compared with LZn group or MZn group. Cytosolic superoxide dismutase activity was the highest in HZn group, while thiobarbituric acid reactive substance level for lipid peroxidation was the highest in LZn group. There was no difference in number of PCNA-positive proliferative cells among the groups. TUNEL-positive apoptotic cells were increased in HZn group compared with LZn group. The HZn group exhibited a decrease of beta-catenin immunostaining areas compared with the LZn or MZn group. These findings indicate that dietary zinc might exert a protecting effect against colon carcinogenesis by inhibiting the development of ACF in the mice.
Aberrant Crypt Foci ; Animals ; Azoxymethane ; beta Catenin ; Colon ; Colorectal Neoplasms ; Cytosol ; Developed Countries ; Dextrans ; Lipid Peroxidation ; Mice ; Mice, Inbred ICR ; Mucous Membrane ; Sodium ; Sulfates ; Superoxide Dismutase ; Thiobarbiturates ; Zinc

OBJECTIVETo observe the effects of astragalosides on content of rat liver microsomal cytochrome P450 (CYP450), activity of glutathione-S-transferase (GST) and dimethylhydrazine (DMH)-induced aberrant crypt foci formation in rats.

METHODSForty SD rats were randomly and equally divided into control group, Astragalus group, DMH group, Astragalus+DMH group. The animals were killed by off neck and the colorectal and liver tissues taken after treatment with astragalosides and dimethyl hydrazine . The colorectal tissues were stained by methylene blue, ACFs observed and counted, and liver microsomes isolated by differential centrifugation. The total enzyme content was detected by using differential spectrometry for carbon monoxide reduction. The glutathione (GSH) level was detected by using spectrophotometry to reflect the activity of the GST.

RESULTSThe numbers of ACF and large ACF in Astragalus+DMH group were more significantly decreased than the DMH group (P<0.05). Compared with the control group, Astragalus group and Astragalus+DMH group, CYP450 level was decreased significantly, and GST activity increased significantly in DMH group (P<0.05).

CONCLUSIONAstragalosides might reduce the number of colorectal aberrant crypt foci induced by dimethylhydrazine possibly by reducing the content of hepatic CYP450 and increasing GST activity in rats.

Aberrant Crypt Foci ; metabolism ; Animals ; Colorectal Neoplasms ; Dimethylhydrazines ; toxicity ; Male ; Microsomes, Liver ; drug effects ; enzymology ; metabolism ; Rats ; Rats, Sprague-Dawley ; Saponins ; pharmacology

We investigated the effect of zinc on the formation of colonic aberrant crypt foci induced by azoxymethane (AOM) followed by dextran sodium sulfate (DSS) in mice with high iron diet (HFe; 450 ppm iron). Six-week old ICR mice were fed on high iron diets with combination of three different levels of zinc in diets, low-zinc (LZn; 0.01 ppm), medium-zinc (MZn; 0.1 ppm), and high-zinc (HZn; 1 ppm) for 12 weeks. Animals were received weekly intraperitoneal injections of AOM (10 mg/kg B.W. in saline) for 3 weeks followed by 2% DSS (molecular weight 36,000~50,000) in the drinking water for a week. To confirm the iron storage in the body, the hepatic iron concentration has been determine chemically and compared with histological assessment visualized by Prussian blue reaction. Aberrant crypt (AC) and aberrant crypt foci (ACF) were analyzed in the colonic mucosa of mouse fed high dietary iron. Superoxide dismutase (SOD) activity and thiobarbituric acid-reactive substances (TBARS) level were also investigated. Apoptosis in the preneoplastic lesion was determined by terminal deoxynucleotidyl transferase-mediated dUTP nickend labeling (TUNEL). In addition, immunohistochemistry of beta-catenin was also performed on the mucous membrane of colon. The number of large ACF (> or = 4 AC/ACF), which possess greater tumorigenic potential, was significantly lower in MZn and HZn groups compared with LZn group. Cytosolic SOD activity in the liver was significantly higher in HZn group compared with LZn group. Hepatic MDA level was decreased significantly in HZn group compared with MZn and LZn groups. Apoptotic index was significantly higher in HZn group. Taken together, these findings indicate that dietary zinc might exert a protective effect against colonic preneoplastic lesion induced by AOM/DSS in ICR mice with high iron status, and suggest that dietary supplement of zinc might play a role in suppressing colon carcinogenesis in mice.
Aberrant Crypt Foci ; Animals ; Apoptosis ; Azoxymethane ; beta Catenin ; Colon ; Cytosol ; Dextrans ; Diet ; Dietary Supplements ; Drinking Water ; Ferrocyanides ; Immunohistochemistry ; Injections, Intraperitoneal ; Iron ; Iron Overload ; Iron, Dietary ; Liver ; Mice ; Mice, Inbred ICR ; Mucous Membrane ; Prussian Blue Reaction ; Sodium ; Sulfates ; Superoxide Dismutase ; Zinc