• Title, Summary, Keyword: Caco-2

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Transport of Transferrin-Horseradish Peroxidase Conjugate Through Cultured Caco-2 Cell Monolayer (배양 Caco-2 세포 단층막 실험계에서 트란스페린과 옥시다아제효소 포합체의 세포막투과)

  • Kim, Dong-Chool;Kim, Jie-Hae
    • Journal of Pharmaceutical Investigation
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    • v.29 no.4
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    • pp.287-293
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    • 1999
  • Transport study of horseradish peroxidase and transferrin-horseradish peroxidase conjugate was performed using an in vitro Caco-2 cell cultured monolayer grown on a polycarbonate membrane of $Transwell^{\circledR}$, Horseradish peroxidase was not transported across Caco-2 cell monolayer. Transferrin-horseradish peroxidase conjugate was transported through Caco-2 cell monolayer. The apparent membrane permeability coefficient $(P_{app})$ of transferrin horseradish peroxidase conjugate was $6.54{\times}10^{-7}\;cm/sec$. The $P_{app}$ value of transferrin-horseradish peroxidase conjugate across Caco-2 cell monolayer was increased to $11.9{\times}10^{-7}\;cm/sec$ in the presence of $50\;{mu}g/ml$ brefeldin-A. These results suggest the transferrin receptor mediated transcytosis of transferrin-horseradish peroxidase conjugate across Caco-2 cell monolayer.

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Growth Stimulation and Inhibition of Differentiation of the Human Colon Carcinoma Cell Line Caco-2 with an Anti-Sense Insulin-Like Growth Factor Binding Protein-3 Construct

  • YoonPark, Jung-Han
    • BMB Reports
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    • v.32 no.3
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    • pp.266-272
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    • 1999
  • The insulin-like growth factor (IGF) system consisting of IGF-I, IGF-II, IGF-receptors, and IGF-binding proteins (IGFBP) regulates the proliferation of a variety of cancer cell types. To examine whether a decrease in endogenous IGFBP-3 stimulates proliferation or inhibits differentiation, Caco-2 cells, a human colon adenocarcinoma cell line, were stably transfected with an anti-sense IGFBP-3 expression construct or pcDNA3 vector as control. Accumulation of IGFBP-3 mRNA and secretion of IGFBP-3 into serum-free conditioned medium, 9 days after plating, were significantly lower in Caco-2 cell clones transfected with anti-sense IGFBP-3 cDNA compared to the controls. The anti-sense clones grew at a similar rate to the controls for 8 days after plating, but achieved a higher final density between days 10 and 12. The levels of sucrase-isomaltase mRNA, a marker of enterocyte differentiation of Caco-2 cells, were lower in the anti-sense clones examined on day 9. In conclusion, proliferation of Caco-2 cells can be stimulated by lowering endogenously-produced IGFBP-3.

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Adhesion of Bifidobacteria to Caco-2 Cells and in Relation to Cell Surface Hydrophobicity (비피도박테리아의 Caco-2 세포에 대한 부착성과 세포 표면 소수성)

  • Lim, Kwang-Sei;Huh, Chul-Sung
    • Food Science of Animal Resources
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    • v.26 no.4
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    • pp.497-502
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    • 2006
  • The adhesion of 16 bifidobacterial strains, including 10 isolates from Korea infants, to Caco-2 cells and their cell surface hydrophobicity were tested. The results of adhesion and cell surface hydrophobicity of for various bifidobacterial strains were obtained and correlations between adhesion and hydrophobicity were strain-dependent properties. Any correlations between species of tested strains were not observed. Among the tested strains, Bifidobacterim longum D6, B. longum H4, B. thermophilum ATCC 25525, B. suis ATCC 27533, and B. animalis subsp. lactis BB12 had higher adherent properties and B. bifidum B3, B. longum D6, B. longum stronger hydrophobicity, respectively. Due to the strain-dependant correlation between adhesion to Caco-2 cells and cell surface hydrophobicity of bifidobacteria, these results provide a possible method for preliminary selection of bifidobacteria potentially adherent to Caco-2 cells by means of cell surface hydrophobic properties.

Comparison of Caco-2 and MDCK Cells As an In-Vitro ADME Screening Model (In-Vitro 흡수특성 검색모델로서 Caco-2 및 MDCK 세포배양계의 특성 비교 평가)

  • Go, Woon-Jung;Cheon, Eun-Pa;Han, Hyo-Kyung
    • Journal of Pharmaceutical Investigation
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    • v.38 no.3
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    • pp.183-189
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    • 2008
  • The present study compared the feasibility of Caco-2 and MDCK cells as an efficient in-vitro model for the drug classification based on Biopharmaceutics Classification System (BCS) as well as an in-vitro model for drug interactions mediated by P-gp inhibition or P-gp induction. Thirteen model drugs were selected to cover BCS Class I{\sim}IV$ and their membrane permeability values were evaluated in both Caco-2 and MDCK cells. P-gp inhibition studies were conducted by using vinblastine and verapamil in MDCK cells. P-gp induction studies were also performed in MDCK cells using rifampin and the P-gp expression level was determined by western blot analysis. Compared to Caco-2 cells, MDCK cells required shorter period of time to culture cells before running the transport study. Both Caco-2 and MDCK cells exhibited the same rank order relationship between in-vitro permeability values and human permeability values of all tested model compounds, implying that those in-vitro models may be useful in the prediction of human permeability (rank order) of new chemical entities at the early drug discovery stage. However, in the case of BCS drug classification, Caco-2 cells appeared to be more suitable than MDCK cells. P-gp induction by rifampin was negligible in MDCK-cells while MDCK cells appeared to be feasible for P-gp inhibition studies. Taken all together, the present study suggests that Caco-2 cells might be more applicable to the BCS drug classification than MDCK-cells, although MDCK cells may provide some advantage in terms of capacity and speed in early ADME screening process.

Adhesion of Kimchi Lactobacillus Strains to Caco-2 Cell Membrane and Sequestration of Aflatoxin B1 (김치 유산균의 Caco-2 세포막 부착성 및 Aflatoxin B1 제거 효과)

  • Lee, Jeongmin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.5
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    • pp.581-585
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    • 2005
  • Five lactic acid bacteria (LAB) including 2 Lactobacillus strains isolated from Kimchi were evaluated to determine the binding ability to Caco-2 cells and $AFB_1$. LAB were divided into three different groups ; viable, heat-treated, and acid-treated cells. In the radioactive-labeling assay for bound cell counting, viable Lactobacillus Plantarum KCTC 3099 showed the higher adhesion to Caco-2 cells with the binding capacity of $39.2\%$, which was $149\%$ higher than Lactobacillus rhamnosus GG as a positive control. Leuconostoc mesenteroids KCTC 3100 showed the similar binding ability to L. rhamnosus GG. After 1 hour incubation at $37^{\circ}C$ with $AFB_1$, viable L. Planterum KTCC 3099 removed the toxin by $49.8\%$, which was similar level to L. rhamnosus GG. Both heat- and acid-treated groups showed high binding effect but acid-treated group was more effective for both Caco-2 cell binding and $AFB_1$ removal than the other. These results indicate that components of bacterial cell wall might be involved in tile binding to intestinal cells and toxins.

Inhibition of Interleukin-1α-induced Intestinal Epithelial Tight Junction Permeability by Curcumin Treatment in Caco-2 Cells in Caco-2 Cells (Caco-2 세포에서 커큐민 처리에 의한 IL-1α로 유도된 소장 상피세포의 tight junction 투과성 저해)

  • Kim, Choon Young
    • Journal of Life Science
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    • v.26 no.9
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    • pp.1082-1087
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    • 2016
  • The intestinal tight junction (TJ) plays an important role as a paracellular barrier. Impaired TJ permeability and enhanced proinflammatory cytokine production are crucial pathophysiological mechanisms in inflammatory bowel diseases (IBDs). Although proinflammatory cytokines, tumor necrosis factor-alpha and interluekin-1 beta, which are markedly increased in IBD patients, have been reported to increase intestinal TJ permeability, the role of interleukin-1 alpha (IL-1α) in the TJ has not been studied. Phytochemicals could prevent proinflammatory cytokine-caused TJ alteration. Curcumin (CCM), a biologically active component of turmeric, has a strong anti-inflammatory activity. The purpose of this study was to elucidate the effect of IL-1α on intestinal epithelial TJ permeability and the role of CCM in IL-1α′s action on TJ in an in vitro intestinal epithelial system, Caco-2 monolayers. The TJ integrity of Caco-2 monolayers was estimated by measuring the flux of FITC-labeled dextran and transepithelial electrical resistance (TEER). Apical IL-1α (100 ng/ml) treatment elevated TJ permeability and suppressed TEER of Caco-2 monolayers. Pretreatment with CCM (20 μM) for 30 min significantly inhibited IL-1α-induced TJ alterations, such as increased TJ permeability and decreased in TEER values. These results demonstrated that IL-1α-induced increases in Caco-2 TJ permeability and CCM blocked the action of IL-1α in the TJ.

Effect of Particle Size of Zinc Oxides on Cytotoxicity and Cell Permeability in Caco-2 Cells

  • Chang, Hyun-Joo;Choi, Sung-Wook;Ko, Sang-Hoon;Chun, Hyang-Sook
    • Preventive Nutrition and Food Science
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    • v.16 no.2
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    • pp.174-178
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    • 2011
  • The cell permeability and cytotoxic effects of different-sized zinc oxide (ZnO) particles were investigated using a human colorectal adenocarcinoma cell line called Caco-2. Morphological observation by scanning electron microscopy revealed that three zinc oxides with different mean particle sizes (ZnO-1, 20 nm; ZnO-2, 90~200 nm; ZnO-3, $1\sim5\;{\mu}m$) tended to aggregate, particularly in the case of ZnO-1. When cytotoxicities of all three sizes of zinc oxide particles were measured at concentration ranges of $1\sim1000\;{\mu}g$/mL, significant decreases in cell viability were observed at concentrations of $50\;{\mu}g$/mL and higher. Among the three zinc oxides, ZnO-1 showed the lowest viability at $50\;{\mu}g$/mL in Caco-2 cells, followed by ZnO-2 and ZnO-3. The permeate concentration of ZnO-1 from the apical to the basolateral side in the Caco-2 model system after four hours was about three-fold higher than that of either ZnO-2 or ZnO-3. These results demonstrated that ZnO-1, with a 20 nm mean particle size, had poorer viability and better permeability in Caco-2 cells than ZnO-2 and ZnO-3.

Inhibitory Effect of Globefish Homogenate on the Growth of Caco-2 Human Colorectal Cancer Cells (복어 균질액의 Caco-2 인간 결장직장암세포 성장 억제 효과에 대한 연구)

  • Kim, Junghoon;Chung, Gujune;Kim, Jungho
    • KSBB Journal
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    • v.32 no.3
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    • pp.212-217
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    • 2017
  • Colorectal cancer is a leading cause of cancer mortality worldwide. Many studies show that most cases of human colorectal cancer arise from adenomatous polyps, which are usually dysplastic, nonmalignant precursor lesions; however, accumulation of multiple somatic mutations leads some to develop into advanced adenoma, which ultimately progresses to an invasive colorectal cancer. Notwithstanding the efforts made to improve chemotherapy, most colorectal cancers are unresponsive to this form of treatment, and malignant colorectal cancers remain incurable. To reduce the incidence of colorectal cancer mortality, further studies to improve colorectal cancer treatment are needed. Here, we show that Globefish homogenate suppresses the growth of Caco-2 human colorectal cancer cells, and that the homogenate inhibits Caco-2 cell proliferation in a dose-dependent manner. These data suggest that Globefish homogenate may suppress colorectal cancer development.

Evaluating the Regulation of P-glycoprotein by Phytochemicals Using Caco-2 Cell Permeability Assay System

  • Choi, Ran Joo;Kim, Yeong Shik
    • Natural Product Sciences
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    • v.20 no.1
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    • pp.1-6
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    • 2014
  • P-glycoprotein (P-gp) is a permeability glycoprotein also known as multidrug resistance protein 1 (MDR1). P-gp is an ATP-binding cassette (ABC) transporter that pumps various types of drugs out of cells. These transporters reduce the intracellular concentrations of drugs and disturb drug absorption. The Caco-2 cell permeability assay system is an effective in vitro system that predicts the intestinal absorption of drugs and the functions of enzymes and transporters. Rhodamine-123 (R-123) and digoxin are well-known P-gp substrates that have been used to determine the function of P-gp. Efflux of P-gp substrates by P-gp has been routinely evaluated. To date, a number of herbal medicines have been tested with Caco-2 cell permeability assay system to assess bioavailability. There are growing efforts to find phytochemicals that potentially regulate P-gp function. The Caco-2 cell permeability assay system is a primary strategy to search for candidates of P-gp inhibitors. In this mini review, we have summarized the P-gp modulation by herbal extracts, decoctions or single components from natural products using Caco-2 cell permeability assays. Many natural products are known to regulate P-gp and herbal medicines could be used in combination with conventional drugs to enhance bioavailability.

Degradation of the Transcription Factors NF-${\kappa}B$, STAT3, and STAT5 Is Involved in Entamoeba histolytica-Induced Cell Death in Caco-2 Colonic Epithelial Cells

  • Kim, Kyeong Ah;Min, Arim;Lee, Young Ah;Shin, Myeong Heon
    • The Korean Journal of Parasitology
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    • v.52 no.5
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    • pp.459-469
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    • 2014
  • Entamoeba histolytica is a tissue-invasive protozoan parasite causing dysentery in humans. During infection of colonic tissues, amoebic trophozoites are able to kill host cells via apoptosis or necrosis, both of which trigger IL-8-mediated acute inflammatory responses. However, the signaling pathways involved in host cell death induced by E. histolytica have not yet been fully defined. In this study, we examined whether calpain plays a role in the cleavage of pro-survival transcription factors during cell death of colonic epithelial cells, induced by live E. histolytica trophozoites. Incubation with amoebic trophozoites induced activation of m-calpain in a time- and dose-dependent manner. Moreover, incubation with amoebae resulted in marked degradation of STAT proteins (STAT3 and STAT5) and NF-${\kappa}B$ (p65) in Caco-2 cells. However, $I{\kappa}B$, an inhibitor of NF-${\kappa}B$, was not cleaved in Caco-2 cells following adherence of E. histolytica. Entamoeba-induced cleavage of STAT proteins and NF-${\kappa}B$ was partially inhibited by pretreatment of cells with a cell-permeable calpain inhibitor, calpeptin. In contrast, E. histolytica did not induce cleavage of caspase-3 in Caco-2 cells. Furthermore, pretreatment of Caco-2 cells with a calpain inhibitor, calpeptin (but not the pan-caspase inhibitor, z-VAD-fmk) or m-calpain siRNA partially reduced Entamoeba-induced DNA fragmentation in Caco-2 cells. These results suggest that calpain plays an important role in E. histolytica-induced degradation of NF-${\kappa}B$ and STATs in colonic epithelial cells, which ultimately accelerates cell death.