alcian-blue and Colonic-Neoplasms

Colon cancers develop after accumulation of multiple genetic and epigenetic alterations in colon epithelial cells. To shed light on global changes in gene expression of colon cancers and to gain further insight into the molecular mechanisms underlying colon carcinogenesis, we have conducted a comprehensive microarray analysis of mRNA using a rat colon cancer model with the food-borne carcinogen, 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP). Of 8749 genes or ESTs on a high density oligonucleotide microarray, 27 and 46 were over- and underexpressed, respectively, by > or =3-fold in colon cancers in common in two rat strains with distinct susceptibility to PhIP carcinogenesis. For example, genes involved in inflammation and matrix proteases and a cell cycle regulator gene, cyclin D2, were highly expressed in colon cancers. In contrast, genes encoding structural proteins, muscle-related proteins, matrix-composing and mucin-like proteins were underexpressed. Interestingly, a subset of genes whose expression is characteristic of Paneth cells, i.e. the defensins and matrilysin, were highly overexpressed in colon cancers. The presence of defensin 3 and defensin 5 transcripts in cancer cells could also be confirmed by in situ mRNA hybridization. Furthermore, Alcian blue/periodic acid Schiff base (AB-PAS) staining and immunohistochemical analysis with an anti-lysozyme antibody demonstrated Paneth cells in the cancer tissues. AB-PAS-positive cells were also observed in high grade dysplastic aberrant crypt foci, which are considered to be preneoplastic lesions of the colon. Our results suggest that Paneth cell differentiation in colon epithelial cells could be an early morphological change in cryptic cells during colon carcinogenesis.

Topics: Alcian Blue; Animals; Carcinogens; Cell Differentiation; Cell Line, Tumor; Colonic Neoplasms; Cyclin D2; Cyclins; Defensins; Expressed Sequence Tags; Gene Expression Regulation, Neoplastic; Humans; Imidazoles; Immunohistochemistry; Muramidase; Nucleic Acid Hybridization; Oligonucleotide Array Sequence Analysis; Oligonucleotides; Paneth Cells; Rats; Rats, Inbred F344; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger

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 beta-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 beta-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.

Topics: 1,2-Dimethylhydrazine; Alcian Blue; Animals; beta Catenin; Biomarkers, Tumor; Carcinogens; Colonic Neoplasms; Cytoskeletal Proteins; Image Processing, Computer-Assisted; Immunohistochemistry; Male; Mucins; Precancerous Conditions; Rats; Rats, Inbred F344; Staining and Labeling; Trans-Activators

Although many colon cancer cell lines are available for study, few of them exhibit differentiated properties. When cultured in medium containing fetal bovine serum, WiDr cells (WiDr-FBS) show an undifferentiated phenotype: growth as a multilayer of cells adherent to plastic and lack of polarization, brush border, and mucin vacuoles. In contrast, WiDr cells cultured in a chemically-defined serum-free medium containing insulin, transferrin and selenium (WiDr-ITS) grow as clusters of nonadherent cells with abundant desmosomes and tight junctions, microvilli and electron-lucid vacuoles. As WiDr-FBS cells, WiDr-ITS are not polarized. WiDr-ITS cells show a marked enhancement in mucin synthesis as demonstrated by: periodic acid-Schiff and Alcian blue stains, electron microscopy, immunohistochemistry using monoclonal antibodies (MAbs) reactive with mucin-associated epitopes, immune electron microscopy and immunochemical analysis using Western blots. In comparison with WiDr-FBS cells, WiDr-ITS cells showed strong expression of Tn, sialyl-Tn, blood group A and CEA. When mouse MAbs were used, higher levels of the MUCI gene product were detected in WiDr-ITS than in WiDr-FBS cells. The full spectrum of phenotypic changes was observed after I month of culture in ITS medium, and transfer of WiDr-ITS cells to FBS medium was accompanied by a partial phenotypic reversal, suggesting that these phenotypic changes result from an adaptative--rather than selective--process.

Topics: Alcian Blue; Antibodies, Monoclonal; Antigens; Blood Group Antigens; Blotting, Western; Cell Division; Colonic Neoplasms; Culture Media; Histocytochemistry; Immunohistochemistry; Microscopy, Electron; Microscopy, Immunoelectron; Mucins; Periodic Acid-Schiff Reaction; Phenotype; Staining and Labeling; Tumor Cells, Cultured

The acid mucins contained in goblet cells of the descending colon of 34 male Sprague-Dawley rats were histochemically labeled by Alcian blue pH 2.5 and quantified in an image analyzer (Cortex Controller). Twenty-two of these 34 rats were treated with 1,2-dimethylhydrazine (DMH) suspended in EDTA solution as a stabilizing agent and the remaining 12 rats with EDTA only. Of the 22 DMH-treated rats, 11 had concomitantly an adenocarcinoma elsewhere in the colon and the remaining 11 rats had no colonic tumors despite DMH treatment. The results indicated that Alcian blue-positive areas occupied 34.5% of the mucosa of the descending colon in tumor-bearing rats, and 35.2% in non-tumor-bearing DMH-treated rats. For EDTA-treated rats the percentage of mucosa occupied by Alcian blue-positive cells was 48.1%. The difference between DMH-treated rats (with or without tumors) and EDTA-treated rats was significant (p less than 0.001). These results suggest that the decrease of Alcian blue areas is related to the protracted treatment with DMH. Whether the decrease in acid mucins is induced by the carcinogen per se or whether it represents a true biochemical premalignant change at the cytoplasmic level remains to be elucidated.

Topics: 1,2-Dimethylhydrazine; Adenocarcinoma; Alcian Blue; Animals; Carcinogens; Colon; Colonic Neoplasms; Dimethylhydrazines; Female; Intestinal Mucosa; Mucins; Rats; Rats, Inbred Strains; Reference Values

Five cases of primary mucinous adenocarcinomas of the lung with signet-ring cells were studied with regard to clinical, pathologic, and prognostic implications and compared with the signet-ring cell adenocarcinomas of extrapulmonary sites. The patients ranged in age from 55 to 74 years, with a mean age of 67.8 years. There were three men and two women. Histologically, three cases were usual adenocarcinomas and two were bronchioloalveolar carcinomas. The percentage of signet-ring cells ranged from 10% to 50%, with a mean of 22% and a median of 20%. Therapy included lobectomy, radiation, and chemotherapy. Three of five patients died of their disease within 9 months and two patients showed no evidence of disease 5 months after presentation. Routine histology showed no significant differences between the signet-ring cells of any of the tumors; however, by special histochemistry, tumors originating from lung, stomach, and colon showed a more intense reaction with alcian blue stain than tumors from nose, breast, or bladder. Contrary to a previous report, we found no increase in sulfated acid mucins in these five cases of lung tumor. We also were unable to demonstrate a qualitative or quantitative difference between mucopolysaccharides produced by lung, stomach, or colon tumors. Although rare, mucinous adenocarcinoma of the lung with signet-ring cells can exist as a primary tumor.

Topics: Adenocarcinoma, Mucinous; Aged; Alcian Blue; Colonic Neoplasms; Female; Histocytochemistry; Humans; Lung Neoplasms; Male; Middle Aged; Mucins; Periodic Acid-Schiff Reaction; Staining and Labeling; Stomach Neoplasms

In a preliminary study, we assessed 10 lectins for the identification of dysplasia in colectomy specimens from patients with ulcerative colitis. Peanut agglutinin (PNA) binding was found in all cases of dysplasia. In the main study the relationship between PNA staining, high iron-diamine/alcian blue (HID-AB) histochemistry, and dysplasia was investigated in 115 pre-operative colonoscopic biopsies and the subsequent resection specimens from patients with ulcerative colitis complicated by carcinoma (n = 6) and patients undergoing proctocolectomy for failure of medical management (n = 8). Peanut lectin was of no value in the assessment of pre-malignant changes or cancer risk. However, the HID-AB stain appears to clarify the interpretation of less severe pre-malignant changes and may be usefully applied to the interpretation of colonoscopic biopsies for cancer surveillance in ulcerative colitis.

Topics: Alcian Blue; Colitis, Ulcerative; Colonic Neoplasms; Histocytochemistry; Humans; Lectins; Mucins; Peanut Agglutinin; Risk

An Alcian blue staining, perchloric acid-soluble, antigenic acidic mucosubstance (GOA) was purified from human gastric signet ring cell carcinoma with DEAE-cellulose chromatography, Seqhadex G-200 and preparative polycrylamide gel electrophoresis. Specific antisera were raised which reacted in indirect immunoenzyme histology with normal goblet cells of the small and large intestine and with goblet cells of intestinalized gastric mucosa. In surgical resection specimens of the stomach (n = 100) and of the colon (n = 19) 3 gastric signet ring cell carcinomas and 3 colonic colloidal adenocarcinomas stained for GOA, demonstrating an immunochemical relationship with normal intestinal goblet cells.

Topics: Adenocarcinoma, Mucinous; Alcian Blue; Antigens, Neoplasm; Colonic Neoplasms; Humans; Stomach Neoplasms