• Journal of Life Science
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• v.26 no.11
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• pp.1313-1319
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• 2016

Cancer is a complex disease heterogeneously composed of various types of cells including cancer stem-like cells responsible for relapse and chemoresistance in the tumor microenvironment. The conventional two-dimensional cell culture-based platform has critical limitations for representing the heterogeneity of cancer cells in the three-dimensional tumor niche in vivo. To overcome this insufficiency, three-dimensional cell culture methods in a scaffold-dependent or -free physical environment have been developed. In this study, we improved and simplified the HCT-8 colon cancer cell-based spheroid culture protocol and evaluated the relationship between cancer stemness and responses of chemosensitivity to 5- Fluorouracil (5-FU), a representative anticancer agent against colon cancer. Supplementation with defined growth factors in the medium and the culture dish of the regular surface with low attachment were required for the formation of constant-sized spheroids containing $CD44^+$ and $CD133^+$ colon cancer stem cells. The chemo-sensitivities of $CD44^+$ cancer stem cells in the spheroids were much lower than those of $CD44^-$ non-stem-like cancer cells, indicating that the chemoresistance to 5-FU is due to the stemness of colon cancer cells. Taken together, the inflammation and oncogenic gut environment-sensitive HCT-8 cell-based colon cancer spheroid culture and comparative evaluation using the simplified model would be an efficient and applicable way to estimate colon cancer stemness and pharmaceutical response to anticancer drugs in the realistic tumor niche.

• The Korean Journal of Physiology and Pharmacology
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• v.2 no.4
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• pp.521-527
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• 1998

An important property of the intestine is the ability to secrete fluid. The intestinal secretion is regulated by a number of substances including vasoactive intestinal peptide (VIP), ATP and different inflammatory mediators. One of the most important secretagogues is adenosine during inflammation. However, the controversy concerning the underlying mechanism of adenosine-stimulated $Cl^-$ secretion in intestinal epithelial cells still continues. To investigate the effect of adenosine on $Cl^-$ secretion and its underlying mechanism in the rabbit colon mucosa, we measured short circuit current ($I_{SC}$) under automatic voltage clamp with DVC-1000 in a modified Ussing chamber. Adenosine, when added to the basolateral side of the muocsa, increased $I_{SC}$ in a dose-dependent manner. The adenosine-stimulated $I_{SC}$ response was abolished when $Cl^-$ in the bath solution was replaced completely with gluconate. In addition, the $I_{SC}$ response was inhibited by a basolateral Na-K-Cl cotransporter blocker, bumetanide, and by apical $Cl^-$ channel blockers, dephenylamine-2-carboxylate (DPC), 5-nitro-2-(3-phenyl-propylamino)-benzoate (NPPB), glibenclamide. Amiloride, an epithelial $Na^+$ channel blocker, and 4,4-diisothiocyanato-stilbene-2,2-disulphonate (DIDS), a $Ca^{2+}-activated$ $Cl^-$ channel blocker, had no effect. In the mucosa pre-stimulated with forskolin, adenosine did not show any additive effect, whereas carbachol resulted in a synergistic potentiation of the $I_{SC}$ response. The adenosine response was inhibited by 10 ${\mu}M$ H-89, an inhibitor of protein kinase A. These results suggest that the adenosine-stimulated $I_{SC}$ response is mediated by basolateral to apical $Cl^-$ secretion through a cAMP-dependent $Cl^-$ channel. The rank order of potencies of adenosine receptor agonists was $5'-(N-ethylcarboxamino)adenosine(NECA)＞N^6-(R-phenylisopropyl)adenosine(R-$ PIA)＞2-[p-(2-carbonylethyl)-phenyl-ethylamino]-5'-N-ethylcarboxaminoadenosine(CGS21680). From the above results, it can be concluded that adenosine interacts with the $A_{2b}$ adenosine receptor in the rabbit colon mucosa and a cAMP-dependent signalling mechanism underlies the stimulation of $Cl^-$ secretion.

• Korean Journal of Plant Resources
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• v.27 no.3
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• pp.223-228
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• 2014

Magnoflorine, an important compound in Aristolochia, was usually used as an anxiolytic chemical. In this study, the magnoflorine was isolated from Aristolochia and the biological activities such as antioxidant, ${\alpha}$-tyrosinase inhibitory, anti-inflammatory, and anticancer activities were investigated. The magnoflorine showed significant antioxidant activity as a 2,2-diphenyl-2-picryl-hydrazyl (DPPH) free radical scavenger, $50{\mu}g/mL$ of the magnoflorine scavenged about 70.8% of all the free radicals. And it was good at ${\alpha}$-tyrosinase inhibiting, $100{\mu}g/mL$ of the magnoflorine inhibited 36.5% of the tyrosinase. High dosage of magnoflorine inhibited the inflammation production nitric oxide (NO), and the magnoflorine protected the murine macrophage cells (RAW 264.7) from LPS-induced apoptosis. The cell viability of human colon cancer calls (HT-29) was around 100% when treated with different dose of magnoflorine, it's suggesting that magnoflorine had no anticancer effect.

• Archives of Pharmacal Research
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• v.26 no.6
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• pp.433-440
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• 2003

Ulcerative colitis (UC) is an inflammatory bowel disease of unknown etiology limited to the large intestine. The disease is prevalent in industrial societies and is associated with specific ethnic populations. A number of murine models, each focused on distinct aspects of the disease process, were developed over the past 20 years to further our understanding of the pathogenesis of UC. These models have been and remain our best resource for the study of the disorder as a result of their homology to human UC and the ease in which they can be manipulated and examined. This review examines and distills what has been learned from these models and how this information is related back to human UC.

• Proceedings of the PSK Conference
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• 2003.10b
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• pp.80.1-80.1
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• 2003

Intestinal epithelial cells can produce cytokines and chemokines that play an important role in the mucosal immune response. Regulation of this production is important to prevent inflammatory tissue damage. Glycyrrhiza glabra has been shown to inhibit inflammation. The aim of this study was to examine the inhibitory effect of 18- beta-glycyrrhetinic acid, a triterpenoid saponin of Glycyrrhiza glabra, on IL-S production via mitogen-activated protein kinases (MAPKs) and nuclear factor-kappa B (NF-kB) in TNF-alpha-stimulated human colon epithelial cells. (omitted)

• Gut and Liver
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• v.12 no.6
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• pp.682-693
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• 2018

Background/Aims: Intestinal barrier dysfunction is a hallmark of inflammatory bowel diseases (IBDs) such as ulcerative colitis. This dysfunction is caused by increased permeability and the loss of tight junctions in intestinal epithelial cells. The aim of this study was to investigate whether estradiol treatment reduces colonic permeability, tight junction disruption, and inflammation in an azoxymethane (AOM)/dextran sodium sulfate (DSS) colon cancer mouse model. Methods: The effects of $17{\beta}$-estradiol (E2) were evaluated in ICR male mice 4 weeks after AOM/DSS treatment. Histological damage was scored by hematoxylin and eosin staining and the levels of the colonic mucosal cytokine myeloperoxidase (MPO) were assessed by enzyme-linked immunosorbent assay (ELISA). To evaluate the effects of E2 on intestinal permeability, tight junctions, and inflammation, we performed quantitative real-time polymerase chain reaction and Western blot analysis. Furthermore, the expression levels of mucin 2 (MUC2) and mucin 4 (MUC4) were measured as target genes for intestinal permeability, whereas zonula occludens 1 (ZO-1), occludin (OCLN), and claudin 4 (CLDN4) served as target genes for the tight junctions. Results: The colitis-mediated induced damage score and MPO activity were reduced by E2 treatment (p<0.05). In addition, the mRNA expression levels of intestinal barrier-related molecules (i.e., MUC2, ZO-1, OCLN, and CLDN4) were decreased by AOM/DSS-treatment; furthermore, this inhibition was rescued by E2 supplementation. The mRNA and protein expression of inflammation-related genes (i.e., KLF4, NF-${\kappa}B$, iNOS, and COX-2) was increased by AOM/DSS-treatment and ameliorated by E2. Conclusions: E2 acts through the estrogen receptor ${\beta}$ signaling pathway to elicit anti-inflammatory effects on intestinal barrier by inducing the expression of MUC2 and tight junction molecules and inhibiting pro-inflammatory cytokines.

• The Journal of Internal Korean Medicine
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• v.38 no.6
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• pp.944-954
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• 2017

Objectives: This study investigated the anti-inflammatory effects of Jageum-jung extract on Dextran sulfate sodium (DSS-induced) ulcerative colitis in mice. Methods: Ulcerative colitis was induced by DSS in Balb/C male mice. Ten mice were assigned to each of four groups: Ctrl (control), UE (ulcerative colitis-induced), PT (treated with pentasaccharide after induction of ulcerative colitis), and JT (treated with Jageum-jung extract after induction of ulcerative colitis). The effects of Jageum-jung extract were measured by restoration of the length of the intestine, degree of mucosal damage as seen with histochemistry, and changes of p-IkB, iNOS, COX-2, and caspase-3 determined by immunohistochemistry. Results: The recovered intestinal length of the JT group was longer than that of the UE group. In the colon mucosa of JT group, hemorrhagic lesions were reduced, and the mucus barrier was recovered. This group also showed inhibited production of inflammatory enzymes (iNOS, COX-2) through regulation of proinflammatory enzyme (NF-kB, p65) activity in the colon. In addition, caspase 3 activation induced apoptosis. By GC/MS analysis, azetidine was identified. Conclusions: This study confirmed the anti-inflammatory effects of jageum-jung extract, and suggests the possibility of using Jageum-jung extract to treat ulcerative colitis. Further experiments and research on the mechanism of Jageum-jung effects are needed.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.4
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• pp.1521-1525
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• 2014

Background: Human papillomavirus (HPV) is one of the most common sexually transmitted infections worldwide and the association between HPV infection and genital cancers has been well established. This study concerned the possible role of HPV infection in colorectal carcinoma (CRC) in the Iranian population. Materials and Methods: We examined 80 tissues obtained from patients with colorectal cancer consisting of 58 colon cancer samples and 22 rectal cancer samples and 80 tissues from patients with unremarkable pathologic changes as matched controls by sex, study center and anatomical sites. HPV infection and genotypes were detected using nested PCR and sequencing methods, respectively. Results: HPV DNA was detected in 5/80 (6.25%) cases including 1 of 22 (4.54%) patients with rectum cancer and 4 of 58 (6.9%) patients with colon cancer and 1/80 (1.25%) of controls. Furthermore, HPV-18 was detected as the most frequent type and we found no significant correlation between prevalence of HPV infection and anatomical sub- sites. Conclusions: Although a causal relation between human papillomavirus and colorectal cancer was not found through this study, analysis of medical records pointed to a possible role for high- risk types of HPV in increasing the potential of aggressiveness in colorectal cancer. This study shows a particular frequency of HPV genotypes in patients with colorectal cancer in Iran. Since HPV vaccines are limited to a few types of virus, using cohort studies in different geographical zones to screen for patterns of HPV infection in different organs might increase the efficacy and optimization of the current vaccines.

• Biomedical Science Letters
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• v.18 no.4
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• pp.362-368
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• 2012

Enterotoxigenic Bacteroides fragilis (ETBF) is an intestinal commensal bacterium implicated as a risk factor for colon cancer. The key virulence factor is a secreted toxin called B. fragilis toxin (BFT). In this study we used an in vitro bioassay to examine the prevalence of ETBF in colonic washings from patients with colorectal polyps and normal control patients. We found that 9.3% of polyp patients and 10.9% of non-polyp patients harbored ETBF, respectively. A total of nine ETBF clinical isolates were isolated and confirmed to be positive for the BFT gene by PCR analysis and the ability to induce IL-8 secretion in the colonic epithelial cell line HT29/c1. Two of the ETBF clinical strains were characterized further in vitro and in vivo. We found that the two ETBF clinical isolates induced E-cadherin cleavage in HT29/c1 cells and promoted colonic inflammation in C57BL/6 mice. Our results indicate that the prevalence of ETBF in polyp patients were similar in non-polyp patients suggesting that ETBF carriage does not positively correlate to polyp incidence.

• Biomolecules & Therapeutics
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• v.20 no.5
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• pp.457-462
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• 2012

The stem-bark of Kalopanax pictus (KP, family Araliaceae), of which main constituent is kalopanaxsaponin B, has been used for asthma, rhinitis, and arthritis in Chinese traditional medicine. To clarify anticolitic effect of KP, we examined anti-inflammatory effect of KP extract and kalopanaxsaponin B in lipopolysaccharide (LPS)-stimulated peritoneal macrophage and 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced colitic mice. Of KP extracts, KP BuOH-soluble fraction most potently inhibited LPS-induced IL-$1{\beta}$, IL-6 and TNF-${\alpha}$ expression, as well as NF-${\kappa}B$ activation. However, KP BuOH fraction increased IL-10, an anti-inflammatory cytokine. KP BuOH fraction also inhibited colon shortening and myeloperoxidase activity in TNBS-induced colitic mice. KP BuOH fraction also potently inhibited the expression of the pro-inflammatory cytokines, IL-$1{\beta}$, IL-6, and TNF-${\alpha}$ as well as the activation of NF-${\kappa}B$. Kalopanaxsaponin B, a main constituent of KP, inhibited TNBS-induced colonic inflammation, including colon shortening, and TNBS-increased myeloperoxidase activity pro-inflammatory cytokine expression and NF-${\kappa}B$ activation in mice. Based on these findings, KP, particularly its main constituent, kalopanaxsaponin B, may ameliorate colitis by inhibiting NF-${\kappa}B$ pathway.