Indigestible carbohydrates which reduce colon tumour incidence in Min mice may interfere with the local immune response.
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Animals↗
Biomedical subjects
J Menanteau
Publications and source records attributed to J Menanteau.
Cytokine mRNA expression in mouse colon: IL-15 mRNA is overexpressed and is highly sensitive to a fibre-like dietary component (short-chain fructo-oligosaccharides) in an Apc gene manner.
On the basis of studies using the Min mouse model of colon carcinogenesis, we have recently proposed that a fibre-like food (short-chain fructo-oligosaccharides, sc-FOS) fermented in the colon may stimulate a mechanism of cancer immunosurveillance. In the present paper, we have investigated the expression of cytokines as potential effector molecules. Interleukin (IL-)4, IL-5, IL-13, IL-15 and interferon (INF)-gamma mRNAs were detected by a multi-probe ribonuclease protection assay in C57BL/6J and Min mouse colons. IL-15 mRNA expression was significantly amplified (P=0.01) by the sc-FOS-enriched diet in the colon of Min mice.
Animals↗
Only fibres promoting a stable butyrate producing colonic ecosystem decrease the rate of aberrant crypt foci in rats.
BACKGROUND: Dietary fibres have been proposed as protective agents against colon cancer but results of both epidemiological and experimental studies are inconclusive. AIMS: Hypothesising that protection against colon cancer may be restricted to butyrate producing fibres, we investigated the factors needed for long term stable butyrate production and its relation to susceptibility to colon cancer. METHODS: A two part randomised blinded study in rats, mimicking a prospective study in humans, was performed using a low fibre control diet (CD) and three high fibre diets: starch free wheat bran (WB), type III resistant starch (RS), and short chain fructo-oligosaccharides (FOS). Using a randomised block design, 96 inbred rats were fed for two, 16, 30, or 44 days to determine the period of adaptation to the diets, fermentation profiles, and effects on the colon, including mucosal proliferation on day 44. Subsequently, 36 rats fed the same diets for 44 days were injected with azoxymethane and checked for aberrant crypt foci 30 days later. RESULTS: After fermentation had stabilised (44 days), only RS and FOS produced large amounts of butyrate, with a trophic effect in the large intestine. No difference in mucosal proliferation between the diets was noted at this time. In the subsequent experiment one month later, fewer aberrant crypt foci were present in rats fed high butyrate producing diets (RS, p=0.022; FOS, p=0.043). CONCLUSION: A stable butyrate producing colonic ecosystem related to selected fibres appears to be less conducive to colon carcinogenesis.
Animals↗
Influence of bcl-2-related proteins on matrix metalloproteinase expression in a rat glioma cell line.
The expression of bcl-2-related proteins has been shown to be a key element in tumoral malignancy. The degradation of the extracellular matrix (ECM) by specialized matrix metalloproteinases (MMPs) is another major step in tumor invasion and metastasis. We have examined, in a rat glioma cell line A15A5, the effect of the stable transfection of human bcl-2, bax and bcl-xl on MMPs expression. Using a zymographic assay, we found that all transfected cell lines expressed a gelatinase activity which is predominantly associated with MMP-9. In bcl-2 and bcl-xl transfected cells, the transcription of MMP-9 was decreased compared to that of control or bax transfected cells. In addition, in bax transfected A15A5, we observed a down regulation of TIMP-1, the inhibitor of MMP-9. These results suggest that the ratio between MMP-9 and its inhibitor TIMP-1 is tightly controlled in cells overexpressing bcl-2 related proteins (i.e., high ratio in bax transfected A15A5 and low ratio in bcl-2 transfected A15A5). However, MMPs secreted by bcl-2 transfected cells were still capable of hydrolyzing FasL present on human lymphocytes. Our results suggest that the expression of bcl-2 related proteins could participate in the regulation of MMP-9/TIMP-1 in gliomas.
Animals↗
T cell status influences colon tumor occurrence in min mice fed short chain fructo-oligosaccharides as a diet supplement.
We have previously shown that addition of short chain fructo-oligosaccharides (indigestible carbohydrates) to food prevented colon tumors in C57BL/6-Apc(Min/+) mice, a model for human colon cancer. As gut-associated lymphoid tissue was concomitantly developed, we suggested that the immune response generated by this food may interfere with carcinogenesis due to involvement of mucosal cells in the regulation of tissue homeostasis. In the present experiment, we tested whether T cell status may influence colon tumor formation in Min mice fed a food supplement of short chain fructo-oligosaccharides. Min mice depleted of CD4(+) and CD8(+) lymphocytes developed twice as many tumors as immunocompetent mice (0.8 as compared with 0.4, the mean number in 7-week-old Min mice when food supplementation began; P = 0.02). It is concluded that food supplementation with a substrate (a known prebiotic) fermented in the colon may stimulate a mechanism of immunosurveillance that would otherwise remain inefficient.
Animals↗
Short-chain fructo-oligosaccharides reduce the occurrence of colon tumors and develop gut-associated lymphoid tissue in Min mice.
C57BL/6J-Min/+ mice, which are heterozygous for a non-sense mutation in the Apc gene, provide a model for both familial adenomatous polyposis and sporadic colon cancers. In our study, gut tumors and small intestine lymphoid nodules were counted in Min mice fed fiber-enriched diets for 6 weeks. Neither starch-free wheat bran nor resistant starch modified the number of tumors. However, short-chain fructo-oligosaccharides dramatically reduced the incidence of colon tumors and concomitantly developed gut-associated lymphoid tissue. Our experiment shows that short-chain fructo-oligosaccharides counteract advanced stages of colon carcinogenesis, possibly via stimulation of antitumoral immunity by modulation of the colonic ecosystem.
Animals↗
Induction of fibroblast gelatinase B expression by direct contact with cell lines derived from primary tumor but not from metastases.
During cancer progression, tumor cells interact with stromal cells. As a consequence, matrix metalloproteinases are produced that contribute to the degradation of the extracellular matrix. This study used coculture systems to investigate fibroblast interaction with three colon cancer cell lines isolated from a single patient. Cells from primary colorectal carcinoma, but not from corresponding liver or lymph node metastases, induced gelatinase B expression by fibroblasts of different tissue origin. Remarkably, direct cell-cell contact was required for this induction, which occurred at the pretranslational level (as revealed by Northern blot analysis) and was completely blocked by anti-beta1 integrin monoclonal antibody, but only partially blocked by anti-alpha5 or anti-alpha(v). Induction was also inhibited by cytochalasin D, staurosporine, or dexamethasone, suggesting the need, respectively, for an organized actin cytoskeleton, protein kinase C, and AP-1-driven gene transcription. Our data suggest that direct tumor-stromal cell contact is one inductive event involved in matrix metalloproteinase expression by stromal cells.
Animals↗
Altered glycosylation of alpha(s)beta 1 integrins from rat colon carcinoma cells decreases their interaction with fibronectin.
Malignant cell transformation is generally accompanied by changes in their interactions with environing matrix proteins in a way to facilitate their migration and generate invasion. Our results show the binding of rat colon adenocarcinoma PROb cells to fibronectin strongly reduced when compared to normal rat intestine epithelial cells. This decrease was not due to the level of alpha(s)beta 1 integrins expressed at the surface of the cell line. However, beta 1- and alpha(s)-associated subunits appeared to be structurally altered as shown by immunoprecipitation followed by electrophoresis. Pulse chase experiments using 35S methionine evidenced differences in the biosynthesis of beta 1- and alpha(s)associated integrins: normal epithelial IEC18 cells required 16 h for maximal biosynthesis of the completely mature beta 1 subunit, while PROb cells did it within 4-6 h. Studies using endoglycosidases O, H, D, and N glycanase confirmed that the molecular weight alterations were due to abnormal glycosylation and suggested that alpha(s)beta 1 integrins of PROb cells could bear both mature complex and immature high mannose types while IEC18 cells borne only mature complex type oligosaccharidic chains. Treatment of both cell types with castanospermine, an inhibitor of N-glycosylation, reduced the differences observed in their adhesion to the fibronectin without significantly affecting beta 1 receptors expression at the cell surface. These results strongly suggest a role of the glycosylation of beta 1 receptors in the adhesion of rat colon adenocarcinoma PROb cells to fibronectin substrata.
Animals↗
Mineralization of Streptococcus mutans in vitro. An ultrastructural study.
Streptococcus mutans mineralization was studied in vitro, with the use of various metastable calcium phosphate solutions, fluoride-containing or otherwise. Degeneration of bacteria always occurred before their mineralization. After complete mineralization of the cytoplasmic area, growth of crystals was observed both in length and in thickness within the extracellular environment. Composition of the calcifying medium was an essential factor in the S. mutans mineralization process. The present study shows that, contrary to carbonate and magnesium ions that prevent intra- and extracellular deposits, fluoride ions promote crystal growth in the above mentioned mediums. Clinical implications of the role of fluoride, at the plaque level, are foreseen.
Calcification, Physiologic↗
Development of a monoclonal antibody against dentin phosphophoryn: a tool to study odontoblastic activity.
The role played by phosphophoryn, one of the major noncollagenous proteins of dentin extracellular matrix, in the mineralization process has not been fully characterized. The purpose of our work was to produce monoclonal antibodies (MAbs) against dentin phosphophoryn and to test their reactivity with primary culture of odontoblasts. Dentin phosphophoryn (DPP) was extracted after the mechanical dissociation of teeth and dialyzed against guanidine and EDTA solutions followed by CaC1(2) precipitation. These extracts were characterized by SDS-PAGE and staining with Coomassie blue and Stains-All. After immunization of mice with these extracts, we produced MAb 7G4, which reacted with dentin phosphophoryn as revealed by Western blot. MAb 7G4 reactivity was tested against a primary culture of pig odontoblasts, revealing filaments specifically stained by the anti-DPP antibody. This antibody will be of great interest to study the mineralization process and dental pulp reaction after capping with various calcium phosphate materials.
Animals↗
Modulation of extracellular matrix glycoproteins production by in vitro interacting conditions between rat colonic fibroblasts and tumoral cells.
Mixed cultures of fibroblasts with rat colon carcinoma cell lines were used to investigate the production of extracellular matrix glycoproteins. Tumoral cells were shown to influence their production in different ways depending on the cell clone (PROb cells which in vivo produce progressive tumors and REGb cells which produce regressive ones) but also on the relative proportions of stromal and tumoral cells. When fibroblasts were predominant, the REGb cells containing mixture produced higher levels of all protein studied as compared with the PROb cells containing system. When the situation was reversed in favor of tumoral cells, REGb cells containing cocultures still produced more fibronectin, laminin and undulin, but the difference with PROb ones was reduced. On the opposite, cocultures enriched with PROb cells made more entactin and SPARC and approximately equal amounts of tenascin.
Animals↗
Differential adhesion of rat colon carcinoma cells to fibronectin in relation to their tumorigenicity.
We examined the fibronectin-adhesive properties of clones from a rat colonic cell line exhibiting distinct tumorigenicity in a syngeneic host. These cells were originally selected on the basis of differential adhesion to plastic surfaces. The TR cell line, when injected subcutaneously, forms a tumour which grows progressively and gives off metastases, whereas the TS cell line forms a small tumour which regresses within a few weeks. The regression is largely mediated by immunological factors and involves a fibroblastic reaction. REGb, a clone from the TS subline, adhered better to fibronectin or RGDS tetrapeptide than did PROb, a clone from the TR subline. However, there was little binding to the RGD tripeptide with either clone. The degree of adhesion was dependent on time and substrate concentration. After 6 h of incubation, 38% and 55% respectively of PROb and REGb cells bound to plates coated with 10 micrograms/ml fibronectin. Adhesion of both clones to fibronectin was inhibited to various degrees when cells were preincubated with RGDS, GRGDS or GRADSPK peptides, whereas other synthetic peptides such as RGD, GRGD or GRGFSPK were ineffective. Binding experiments using 125I-labelled fibronectin showed 39,000 fibronectin receptor sites on REGb cells but only 17,000 on PROb cells. Flow cytometry analysis using both anti-alpha 5 and anti-beta 1 integrins showed more fibronectin receptor sites on REGb than on PROb cells. Both approaches were in accordance with the higher adhesiveness of the REGb clone to fibronectin.(ABSTRACT TRUNCATED AT 250 WORDS)
Adenocarcinoma↗
Cellular activity of osteoblasts in the presence of hydroxyapatite: an in vitro experiment.
The use of synthetic calcium phosphate as bone substitute calls for the knowledge of the influence on adjacent cells. Effects on monocytes, macrophages, synovial cells and fibroblasts have been largely described in vivo and in vitro but few data are available as concerns osteoblast responses. The present experiments tested the activity of MC3T3-E1, ROS 17/2.8 and mouse calvaria cells cultured in the presence of hydroxyapatite powders. The three osteoblast-like cells were shown to phagocytoze the calcium phosphate particles. As a consequence, they exhibit reduced cell growth and alkaline phosphatase activity. This response was different when compared with other cell types. The osteogenetic function of osteoblastic cells could be involved in these specific effects of hydroxyapatite.
3T3 Cells↗
The influence of calcium phosphate biomaterials on human bone cell activities. An in vitro approach.
An in vitro method is described to assess the influence of synthetic calcium phosphate powders on osteoblast activities. Human osteoblast cell cultures were established from iliac crest. MC3T3-E1, an established osteogenic cell line, was employed as a control. Scanning and transmission electron microscopic observations clearly demonstrated the internalization of particles of calcium phosphate by the two osteoblast cell populations. As a consequence to the phagocytotic process, RNA transcription and protein synthesis were stimulated, as indicated by the measurements of labeled uridine, leucine and proline uptakes. From these data, it is proposed that such an in vitro model, using one of the specific cell types involved in the tissue responses to implants, could be useful to assess the biological response at the cell-biomaterial interaction.
Alkaline Phosphatase↗
Proteins of the mineral compartment of bovine fetal enamel share common antigenic determinants with serum proteins.
Two fractions were separated from the proteins of the mineral compartment of bovine developing enamel on the basis of their affinity for the lectin concanavalin-A. A monoclonal antibody was prepared by the hybridoma technique against the Con-A-binding fraction. This antibody and a commercial polyclonal antibody against bovine serum albumin were used to examine the relationship between those proteins, serum albumin and alpha-2HS glycoprotein, two proteins concentrated within dentin and bone matrices. The Con-A-unbound fraction reacted with the anti-albumin antibody and the antibody against the Con-A-binding fraction recognized the alpha-2HS glycoprotein. These data fully support the presence of significant levels of proteins related to serum components in the mineral compartment of developing enamel matrix.
Animals↗
Effects of synthetic calcium phosphates on the 3H-thymidine incorporation and alkaline phosphatase activity of human fibroblasts in culture.
Gingival fibroblasts were cultured with four different calcium phosphate minerals (hydroxyapatite, whitlockite, beta-tricalcium phosphate, and octocalcium phosphate). 3H-thymidine incorporation into DNA and alkaline phosphatase specific activity were determined after different incubation periods. As a consequence of the phagocytosis of calcium phosphates crystals, we pointed out, compared to control, a stimulation of the rate of 3H-thymidine incorporation and sharp decreases in alkaline phosphatase activity. The magnitude of the alkaline phosphatase activity inhibition was observed to be increased with the solubility of the materials. We propose that the effects of calcium phosphates on alkaline phosphatase and 3H-thymidine incorporation could be calcium-mediated events, resulting from intracellular dissolution of phagocytized materials. We suggest that in vitro determination of 3H-thymidine incorporation and alkaline phosphatase activity, which are highly sensitive tests, could be involved in evaluation procedures of calcium phosphates biomaterials.
Adult↗
Chemical changes in hydroxyapatite biomaterial under in vivo and in vitro biological conditions.
The introduction of a synthetic calcium phosphate into a biological environment is likely to result in surface-mediated chemical events. On the basis of such an assessment, we studied the chemical changes occurring in the mineral after exposure of a synthetic hydroxyapatite ceramic to both in vivo (implantation in human) and in vitro (cell culture) conditions. A small amount of the material was phagocytized but the major remaining part behaved as a secondary nucleator as evidenced by the appearance of a newly formed mineral. Morphologically, the newly formed mineral appeared as tiny crystals precipitated and grown from the surface of the initial synthetic crystals. The density of the additional mineral increased from the periphery to the core of each biomaterial aggregate. Chemically, it was identified by IR spectroscopy as a carbonated apatitic mineral. We propose that the adsorption of biomolecules could inhibit precipitation, accounting for the increasing amount of precipitate from the periphery to the core of the aggregates.
Adult↗
A western-blotting study of enamel glycoproteins in rat experimental fluorosis.
Experimental fluorosis was induced in order to get information on enamel protein glycosylation, using Western-blotting methodology with peroxidase-labelled concanavalin A. Fluoride inhibited amelogenin degradation, especially the production of intermediate forms. Within the non-amelogenin family of proteins there were changes in both the conventionally stainable components and the glycoconjugates revealed by lectin only. Fluoride influenced the whole extracellular processing of enamel proteins including movement between the mineral and non-mineral compartments. A different degradation scheme of enamel proteins, which also affects the glycoconjugates, might be of importance in the properties of the fluorosed enamel surface and its interactions with the oral environment.
Amido Black↗