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Journal of Nutritional Science and... 2019The effect of low-dose of ethanol consumption on the development of colon cancer is unclear. This study aimed to investigate the effects of low-dose ethanol (0.5%, 1%,...
The effect of low-dose of ethanol consumption on the development of colon cancer is unclear. This study aimed to investigate the effects of low-dose ethanol (0.5%, 1%, and 2% [v/v] ethanol in drinking water) for 28 wk on colon tumor incidence in rats injected with 1,2-dimethylhydrazine. Body weight, fluid and food consumption, and the total numbers of colon adenomas (mild-, moderate-, and severe-grade dysplasia) per rat were unaffected by ethanol consumption. However, the numbers of severe-grade dysplasia were significantly reduced by 1% ethanol compared with the control (0% ethanol; -93%) but not by 0.5% and 2% ethanol. Although the numbers of total adenocarcinomas were unaffected, those of total of adenomas and adenocarcinomas together were significantly reduced by 0.5% and 1% ethanol (-39% and -41%, respectively). Intriguingly, real-time PCR assay indicated the abundance of cecal Clostridium leptum (a putative immunosuppressor) was the least in rats received 1% ethanol. Furthermore, 1% ethanol markedly increased colonic mRNA of IL-6, a putative suppressor of regulatory T-cells and cytoprotector. This study provides the first evidence for the potential of 1% ethanol, but not 2% ethanol, to prevent colon tumorigenesis in rats, supporting the J-curve hypothesis of the effect of low-dose alcohol on health. Further, the modulation of C. leptum and expression of IL-6, potentially linking to carcinogenesis, by 1% ethanol may provide an insight into the underlying mechanisms of the anti-colon tumor effect.
Topics: 1,2-Dimethylhydrazine; Adenocarcinoma; Animals; Antineoplastic Agents; Carcinogenesis; Colon; Colonic Neoplasms; Ethanol; Interleukin-6; RNA, Messenger; Rats
PubMed: 31666482
DOI: 10.3177/jnsv.65.443 -
The Journal of Clinical Investigation Mar 1986Dimethylhydrazine (DMH) is a potent procarcinogen with selectivity for the colon. To determine whether alterations in the lipid composition and fluidity of rat colonic...
Dimethylhydrazine (DMH) is a potent procarcinogen with selectivity for the colon. To determine whether alterations in the lipid composition and fluidity of rat colonic brush border membranes existed before the development of DMH-induced colon cancer, rats were injected s.c. with this agent (20 mg/kg body weight per wk) or diluent for 5, 10, and 15 wk. Animals were killed at these time periods and brush border membranes were prepared from proximal and distal colonocytes of each group. The "static" and "dynamic" components of fluidity of each membrane were then assessed, by steady-state fluorescence polarization techniques using limiting hindered fluorescence anisotropy and order parameter values of the fluorophore 1,6 diphenyl-1,3,5-hexatriene (DPH) and fluorescence anisotropy values of DL-2-(9-anthroyl) stearic acid and DL-12-(9-anthroyl) stearic acid, respectively. Membrane lipids were extracted and analyzed by thin-layer chromatography and gas-liquid chromatography. Phospholipid methylation activity in these membranes was also measured using S-adenosyl-L-methionine as the methyl donor. The results of these studies demonstrate that: the lipid composition and both components of fluidity of proximal DMH-treated and control membranes and their liposomes were similar at all time periods examined; at 5, 10, and 15 wk the "dynamic component of fluidity" of distal DMH-treated membranes and their liposomes was found to be higher, similar, and lower, respectively, than their control counterparts; the "static component of fluidity" of distal DMH-treated membranes and their liposomes, however, was similar to control preparations at all three time periods; and alterations in the lipid composition and phospholipid methylation activities appeared to be responsible for these differences in the "dynamic component of fluidity" at these various time periods.
Topics: 1,2-Dimethylhydrazine; Animals; Colon; Dimethylhydrazines; Fatty Acids; Fluorescence Polarization; Intestinal Mucosa; Kinetics; Male; Membrane Fluidity; Membrane Lipids; Methylation; Methylhydrazines; Methyltransferases; Microvilli; Phosphatidylethanolamine N-Methyltransferase; Rats; Time Factors
PubMed: 3949981
DOI: 10.1172/JCI112380 -
Nutrients Aug 2014This work investigated the effects of Vitamin E (VE) on aberrant crypt foci (ACF) incidence, oxidative stress parameters (serum and hepatic VE concentration, and...
This work investigated the effects of Vitamin E (VE) on aberrant crypt foci (ACF) incidence, oxidative stress parameters (serum and hepatic VE concentration, and homocysteine, glutathione (GSH), and malondialdehyde (MDA) levels), and expression of both cyclooxygenase-2 (COX2) and proliferating cellular nuclear antigen (PCNA) in experimental colorectal carcinogenesis. Male Wistar rats received subcutaneous injections of 1,2-dimethylhydrazine (DMH) twice a week, for two weeks (40 mg/kg), except for the Control group. Animals were separated into groups that received different amounts of VE in the diet: 0 IU (0×), 75 IU (recommended daily intake, RDI), 225 IU (3× RDI), or 1500 IU (20× RDI), during (dDMH) or after (aDMH) administration of carcinogen. The 0×dDMH and 3×dDMH groups showed decreased serum VE levels. Hepatic VE concentration was higher in 3×aDMH as compared with the other groups. All the groups, except the Control and the 0×aDMH groups, had reduced GSH levels. The 0×dDMH, 0×aDMH, and 20×aDMH groups exhibited increased MDA levels. The aDMH groups had higher ACF incidence and PCNA expression. The 0×aDMH group presented higher ACF rate, followed by 20×aDMH. Moreover, the 3×aDMH group displayed reduced ACF incidence and COX2 expression. Multivariate analysis revealed that GSH modulated homocysteine levels and COX2. These results suggested that 1500 IU of VE is hazardous, whereas 225 IU of VE has beneficial effects on chemical colorectal carcinogenesis.
Topics: 1,2-Dimethylhydrazine; Aberrant Crypt Foci; Animals; Biomarkers; Carcinogenesis; Carcinogens; Cell Proliferation; Colorectal Neoplasms; Cyclooxygenase 2; Dietary Supplements; Disease Models, Animal; Dose-Response Relationship, Drug; Gene Expression Regulation, Neoplastic; Glutathione; Homocysteine; Immunohistochemistry; Male; Multivariate Analysis; Oxidative Stress; Proliferating Cell Nuclear Antigen; Rats; Rats, Wistar; Recommended Dietary Allowances; Vitamin E; Weight Gain
PubMed: 25123248
DOI: 10.3390/nu6083214 -
European Review For Medical and... Jun 2019The aim of this study was to explore the regulatory mechanism of micro ribonucleic acid (miR)-223 in ulcerative colitis (UC) through the phosphatidylinositol 3-kinase...
OBJECTIVE
The aim of this study was to explore the regulatory mechanism of micro ribonucleic acid (miR)-223 in ulcerative colitis (UC) through the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt)-mammalian target of rapamycin (mTOR) signaling pathway.
MATERIALS AND METHODS
A total of 36 Sprague-Dawley (SD) rats were randomly divided into three groups, including normal group (n=12), model group (n=12) and inhibitor group (n=12). Rats in the normal group received no treatment. Rats in the model group were used to establish a UC model. Meanwhile, rats in the inhibitor group underwent intraperitoneal injection of inhibitor and establishment of the UC model. Subsequently, specimens were obtained for detection. Immunohistochemistry was applied to measure the expression of mTOR. Western blotting was adopted to determine the relative protein expressions of P85, P110 and phosphorylated Akt (p-Akt). Quantitative polymerase chain reaction (qPCR) was used to detect the mRNA expression of miR-223. Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) was utilized to determine cell apoptosis. Furthermore, an enzyme-linked immunosorbent assay (ELISA) was conducted to measure the content of interleukin-1 beta (IL-1β) and IL-6.
RESULTS
Immunohistochemistry showed that the positive expression of mTOR increased remarkably in the model group and inhibitor group when compared with that of the normal group (p<0.05). However, it decreased notably in the inhibitor group when compared with the model group (p<0.05). Western blotting indicated that the protein expressions of P85, P110 and p-Akt in model group and inhibitor group were significantly higher than the ones of the normal group (p<0.05). However, the inhibitor group showed markedly lower relative protein expressions of P85, P110 and p-Akt than the ones of the model group (p<0.05). Compared with the normal group, the expression level of miR-223 was significantly elevated in model group and inhibitor group (p<0.05). However, there was no significant difference in the mRNA expression of miR-233 between the model group and the inhibitor group (p>0.05). The apoptosis rate of the cells increased prominently in the model group and in the inhibitor group when compared with the normal group (p<0.05). However, it was remarkably reduced in the inhibitor group than the model group (p<0.05). In comparison with the normal group, the content of IL-1β and IL-6 was significantly up-regulated in the model group and in the inhibitor group (p<0.05). However, it declined notably in the inhibitor group compared with the model group (p<0.05).
CONCLUSIONS
MiR-223 can trigger cell apoptosis and inflammation in UC by up-regulating the PI3K/Akt-mTOR signaling pathway.
Topics: Animals; Apoptosis; Chromones; Colitis, Ulcerative; Dextran Sulfate; Dimethylhydrazines; Disease Models, Animal; Humans; Interleukin-1beta; Interleukin-6; MicroRNAs; Morpholines; Phosphatidylinositol 3-Kinases; Phosphoinositide-3 Kinase Inhibitors; Proto-Oncogene Proteins c-akt; Rats; Signal Transduction; TOR Serine-Threonine Kinases; Up-Regulation
PubMed: 31210320
DOI: 10.26355/eurrev_201906_18074 -
BMC Cancer Feb 2013Colon cancer stem cells may drive carcinogenesis and account for chemotherapeutic failure. Although many markers for these cells have been proposed, there is no complete...
Expression of LGR-5, MSI-1 and DCAMKL-1, putative stem cell markers, in the early phases of 1,2-dimethylhydrazine-induced rat colon carcinogenesis: correlation with nuclear β-catenin.
BACKGROUND
Colon cancer stem cells may drive carcinogenesis and account for chemotherapeutic failure. Although many markers for these cells have been proposed, there is no complete agreement regarding them, nor has their presence in the early phases of carcinogenesis been characterized in depth.
METHODS
The expression of the putative markers LGR-5 (leucine-rich-repeat-containing G-protein-coupled receptor 5), MSI-1 (Musashi-1) and DCAMKL-1 (doublecortin and calcium/calmodulin-dependent protein kinase-like-1) was studied in normal colon mucosa (NM), in the precancerous lesions Mucin Depleted Foci (MDF) and in macroscopic tumours (adenomas) of 1,2-dimethylhydrazine-treated rats. Co-localization between these markers and nuclear β-catenin (NBC), an attributed feature of cancer stem cells, was also determined. Moreover, since PGE2 could increase NBC, we tested whether short-term treatment with celecoxib, a COX-2 inhibitor (2 weeks, 250 ppm in the diet) could reduce the expression of these markers.
RESULTS
LGR-5 expression in NM was low (Labelling Index (LI): 0.22 ± 0.03 (means ± SE)) with positive cells located mainly at the base of the crypts. Compared to NM, LGR-5 was overexpressed in MDF and tumours (LI: 4.7 ± 2.0 and 2.9 ± 1.0 in MDF and tumours, respectively, P<0.01 compared to NM). DCAMKL-1 positive cells, distributed along the length of normal crypts, were reduced in MDF and tumours. Nuclear expression of MSI-1, located mainly at the base of normal crypts, was not observed in MDF or tumours. In both MDF and tumours, few cells co-expressed LGR-5 and NBC (LI: 1.0 ± 0.3 and 0.4 ± 0.2 in MDF and tumours, respectively). Notwithstanding the lower expression of DCAMKL-1 in tumours, the percentage of cells co-expressing DCAMKL-1 and NBC was higher than in NM (LI: 0.5 ± 0.1 and 0.04 ± 0.02 in tumours and NM, respectively). MSI-1 and NBC co-localization was not observed. Celecoxib did not reduce cells co-expressing LGR-5 and NBC.
CONCLUSIONS
Based on its prevalent localization at the base of normal crypts, as expected for stem cells, and on the overexpression in precancerous lesions and tumours, we support LGR-5, but not MSI-1 or DCAMKL-1, as putative neoplastic stem cell marker. In both MDF and tumours, we identified LGR-5-positive cells co-expressing NBC which could be a subpopulation with the highest stem cell features.
Topics: 1,2-Dimethylhydrazine; Adenoma; Animals; Biomarkers, Tumor; Celecoxib; Cell Nucleus; Cell Transformation, Neoplastic; Colonic Neoplasms; Cyclooxygenase 2; Cyclooxygenase 2 Inhibitors; Dinoprostone; Doublecortin Protein; Doublecortin-Like Kinases; Fluorescent Antibody Technique; Immunohistochemistry; Intestinal Mucosa; Male; Neoplastic Stem Cells; Nerve Tissue Proteins; Precancerous Conditions; Protein Serine-Threonine Kinases; Pyrazoles; RNA-Binding Proteins; Rats; Rats, Inbred F344; Receptors, G-Protein-Coupled; Sulfonamides; Time Factors; beta Catenin
PubMed: 23374535
DOI: 10.1186/1471-2407-13-48 -
International Journal of Molecular... Jun 2022Colorectal cancer (CRC) is one of most common cancers worldwide, with high rates of mortality. Epidemiological findings demonstrate that coffee consumption reduces the...
Colorectal cancer (CRC) is one of most common cancers worldwide, with high rates of mortality. Epidemiological findings demonstrate that coffee consumption reduces the risk of developing CRC by ~13%. In general, in vivo and in vitro findings demonstrate the antiproliferative, antioxidant and proapoptotic effects of brewed coffee or major bioavailable coffee compounds. Thus, it was assessed whether caffeine (CAF) and/or chlorogenic acid (CGA) attenuates the early-stage of chemically induced mouse colon carcinogenesis. Male Swiss mice were submitted to a 1,2-dimethylhydrazine/deoxycholic acid (DMH/DCA)-induced colon carcinogenesis model. These animals received CAF (50 mg/kg), CGA (25 mg/kg) or CAF+CGA (50 + 25 mg/kg) intragastrically for five times/week for ten weeks. CAF+CGA had the most pronounced effects on decreasing epithelial cell proliferation (Ki-67) and increasing apoptosis (cleaved caspase-3) in colonic crypts. This treatment also decreased the levels of proinflammatory cytokines IL-6, IL-17 and TNF-α, and downregulated the oncomiR miR-21a-5p in the colon. Accordingly, the analysis of miR-21a-5p targets demonstrated the genes involved in the negative regulation of proliferation and inflammation, and the positive regulation of apoptosis. Ultimately, CAF+CGA attenuated preneoplastic aberrant crypt foci (ACF) development. Our findings suggest that a combination of coffee compounds reduces early-stage colon carcinogenesis by the modulation of miR-21a-5p expression, highlighting the importance of coffee intake to prevent CRC.
Topics: 1,2-Dimethylhydrazine; Animals; Caffeine; Carcinogenesis; Chlorogenic Acid; Coffee; Colon; Colonic Neoplasms; Male; Mice; MicroRNAs
PubMed: 35682971
DOI: 10.3390/ijms23116292 -
International Journal of Molecular... Sep 2021Colon cancer is accompanied by a decrease of epithelial barrier properties, which are determined by tight junction (TJ) proteins between adjacent epithelial cells. The...
Colon cancer is accompanied by a decrease of epithelial barrier properties, which are determined by tight junction (TJ) proteins between adjacent epithelial cells. The aim of the current study was to analyze the expression of TJ proteins in a rat model of 1,2-dimethylhydrazine (DMH)-induced colorectal cancer, as well as the barrier properties and TJ protein expression of IPEC-J2 cell monolayers after incubation with DMH. Transepithelial electrical resistance and paracellular permeability for sodium fluorescein of IPEC-J2 were examined by an epithelial volt/ohm meter and spectrophotometry. The expression and localization of TJ proteins were analyzed by immunoblotting and immunohistochemistry. In the colonic tumors of rats with DMH-induced carcinogenesis, the expression of claudin-3 and -4 was significantly increased compared to controls. The transepithelial electrical resistance of IPEC-J2 cells increased, while paracellular permeability for sodium fluorescein decreased, accompanied by an increased expression of claudin-4. The increase of claudin-4 in rat colon after chronic DMH exposure was consistent with the acute effect of DMH on IPEC-J2 cells, which may indicate an essential role of this protein in colorectal cancer development.
Topics: 1,2-Dimethylhydrazine; Adenocarcinoma; Animals; Carcinogens; Cell Line; Claudins; Colonic Neoplasms; Electric Impedance; Intestinal Mucosa; Male; Permeability; Rats; Rats, Wistar; Swine; Tight Junction Proteins
PubMed: 34638619
DOI: 10.3390/ijms221910278 -
Nutricion Hospitalaria Feb 2020Introduction: resistant starch (RS) is not completely digested in the human intestine but is fermented in the colon; intestinal pH decreases as short-chain fatty acids... (Comparative Study)
Comparative Study
Introduction: resistant starch (RS) is not completely digested in the human intestine but is fermented in the colon; intestinal pH decreases as short-chain fatty acids are produced. This is beneficial for health, and for preventing and treating rectal colon cancer. Pyrodextrinization and enzymatic hydrolysis are modifications to native starch (NS) that may increase the amount of RS. Objective: the objective of this project was to evaluate the effects of M. cavendish AAA native and both chemically and enzymatically modified starches on tumor markers in rats. Methods: modifications (chemical and enzymatic) were made to M. cavendish AAA NS, and were evaluated in rats with 1,2-DMH. Male Sprague Dawley rats (25) were used, divided into five experimental groups: PC, NC, NS, PI, and ERM. During 4 weeks they received the experimental diet assigned to each group. The PC, NS, PI and ERM groups received 2 weekly s.c. (subcutaneous) injections of 1,2-DMH (40 mg/kg) (third and fourth week). In feces, pH, β-glucuronidase enzyme, and short-chain fatty acids were evaluated, and a histopathological study was performed of the intestine to detect microscopic lesions. Results: the activity of β-glucuronidase decreased (p < 0.05) for NS, PI and ERM vs. PC. The highest proportion of butyric acid was observed in the NS (p < 0.05) vs. NC group. Sixty percent of enteritides were severe in grade in the PC group, and 40 % in the experimental groups. Conclusions: native starch granules resisted pyrodextrinization, but treatment with α-amylase broke the structure of the pyrodextrin granule. According to the treatments given to the rats, as the amount of RS present in the diet increases (NS), the neoplastic cells do not advance beyond the basement membrane, suggesting a possible cell-protective or anticancer effect.
Topics: 1,2-Dimethylhydrazine; Animals; Carcinogens; Colonic Neoplasms; Fatty Acids, Volatile; Feces; Glucuronidase; Hydrogen-Ion Concentration; Hydrolysis; Intestinal Mucosa; Male; Microscopy, Electron; Musa; Polysaccharides; Random Allocation; Rats; Rats, Sprague-Dawley; Starch; alpha-Amylases
PubMed: 31793323
DOI: 10.20960/nh.02706 -
PloS One 2016Doxycycline (DOX) exhibits anti-inflammatory, anti-tumor, and pro-apoptotic activity and is being tested in clinical trials as a chemotherapeutic agent for several...
BACKGROUND
Doxycycline (DOX) exhibits anti-inflammatory, anti-tumor, and pro-apoptotic activity and is being tested in clinical trials as a chemotherapeutic agent for several cancers, including colon cancer.
MATERIALS & METHODS
In the current study, the chemotherapeutic activity of doxycycline was tested in a rat model of colon carcinogenesis, induced by colon specific cancer promoter, 1,2, dimethylhydrazine (DMH) as well as study the effect of DOX-alone on a separate group of rats.
RESULTS
Doxycycline administration in DMH-treated rats (DMH-DOX) unexpectedly increased tumor multiplicity, stimulated progression of colonic tumor growth from adenomas to carcinomas and revealed metastasis in small intestine as determined by macroscopic and histopathological analysis. DOX-alone treatment showed markedly enhanced chronic inflammation and reactive hyperplasia, which was dependent upon the dose of doxycycline administered. Moreover, immunohistochemical analysis revealed evidence of inflammation and anti-apoptotic action of DOX by deregulation of various biomarkers.
CONCLUSION
These results suggest that doxycycline caused chronic inflammation in colon, small intestine injury, enhanced the efficacy of DMH in tumor progression and provided a mechanistic link between doxycycline-induced chronic inflammation and tumorigenesis. Ongoing studies thus may need to focus on the molecular mechanisms of doxycycline action, which lead to its inflammatory and tumorigenic effects.
Topics: 1,2-Dimethylhydrazine; Animals; Body Weight; Carcinogenesis; Caspase 3; Caspase 9; Cell Proliferation; Chronic Disease; Colonic Neoplasms; Cytochromes c; Down-Regulation; Doxycycline; Immunohistochemistry; Inflammation; Intestine, Small; Male; Matrix Metalloproteinase 9; NF-kappa B; Neoplasm Metastasis; Rats, Sprague-Dawley; Tumor Suppressor Protein p53; Up-Regulation; Vascular Endothelial Growth Factor A
PubMed: 26998758
DOI: 10.1371/journal.pone.0151539 -
Brazilian Journal of Medical and... Jun 2011β-ionone (βI), a cyclic isoprenoid, and geraniol (GO), an acyclic monoterpene, represent a promising class of dietary chemopreventive agents against cancer, whose...
β-ionone (βI), a cyclic isoprenoid, and geraniol (GO), an acyclic monoterpene, represent a promising class of dietary chemopreventive agents against cancer, whose combination could result in synergistic anticarcinogenic effects. The chemopreventive activities of βI and GO were evaluated individually or in combination during colon carcinogenesis induced by dimethylhydrazine in 48 3-week-old male Wistar rats (12 per group) weighing 40-50 g. Animals were treated for 9 consecutive weeks with βI (16 mg/100 g body weight), GO (25 mg/100 g body weight), βI combined with GO or corn oil (control). Number of total aberrant crypt foci (ACF) and of ACF ≥4 crypts in the distal colon was significantly lower in the GO group (66 ± 13 and 9 ± 2, respectively) compared to control (102 ± 9 and 17 ± 3) and without differences in the βI (91 ± 11 and 14 ± 3) and βI+GO groups (96 ± 5 and 19 ± 2). Apoptosis level, identified by classical apoptosis morphological criteria, in the distal colon was significantly higher in the GO group (1.64 ± 0.06 apoptotic cells/mm²) compared to control (0.91 ± 0.07 apoptotic cells/mm²). The GO group presented a 0.7-fold reduction in Bcl-2 protein expression (Western blot) compared to control. Colonic mucosa concentrations of βI and GO (gas chromatography/mass spectrometry) were higher in the βI and GO groups, respectively, compared to the control and βI+GO groups. Therefore, GO, but not βI, represents a potential chemopreventive agent in colon carcinogenesis. Surprisingly, the combination of isoprenoids does not represent an efficient chemopreventive strategy.
Topics: Acyclic Monoterpenes; Animals; Anticarcinogenic Agents; Antineoplastic Combined Chemotherapy Protocols; Carcinogens; Colon; Colonic Neoplasms; Dimethylhydrazines; Drug Screening Assays, Antitumor; Intestinal Mucosa; Male; Norisoprenoids; Rats; Rats, Wistar; Terpenes
PubMed: 21445532
DOI: 10.1590/s0100-879x2011007500037