• Title/Summary/Keyword: colon cell line

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$\beta$-Alanine Induced Down-Regulation of the Taurine Transporter Activity in the Human Colon Carcinoma Cell Line (HT-29) (인체 소장상피세포주 모델(HT-29)에서 $\beta$-알라닌이 타우린수송체 활성에 미치는 영향)

  • 박태선;윤미영;정한나;이해미
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.30 no.2
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    • pp.314-319
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    • 2001
  • In the present study, effects of $\beta$-alanine, a known taurine antagonist for its structural similarity, on the adaptive regulation and kinetic behavior of the taurine transporter were investigated in the HT-29, human colon carcinoma cell line. Pretreatment of the cell with $\beta$-alanine(10mM) for varying periods from 3 to 30 hrs significantly reduced the taurine uptake compared to the value for control cells. This decrease in the taurine transporter activity was dependent on the incubation time with $\beta$-alanine, and the maximal down-regulation of the transporter activity was observed in cells pretreated with $\beta$-alanine for 24 hrs (25% of the control value, p<0.01). The taurine transporter appears to bind exclusively with $\beta$-alanine in the HT-29 cells since the same concentration of $\alpha$-alanine added in the culture medium for 24 hrs did not influence the taurine uptake. Kinetic analyses of the taurine transporter activity was performed in the HT-29 cell line with varying taurine concentration (5~60$\mu$M) in the uptake medium. Active taurine uptake was significantly lower in $\beta$-alanine pretreated cells compared to the value for control cells in the range of taurine concentration used in the experiment (p<0.001). The cells pretreated with $\beta$-alanine showed a 50% lower maximal velocity (Vmax, 1.7$\pm$2.0 nmole.mg $protein^{-1}$.$30min^{-1}$), and a 99% higher Michaelis constant (Km, 40.3$\pm$7.6$\mu$M) than the control values (3.3$\pm$1.9 nmole.mg $protein^{-1}$.$30min^{-1}$, and 20.3$\pm$2.1$\mu$M, respectively). These results on kinetic data suggest that $\beta$-alanine induced down-regulation of the taurine transporter activity was associated with decreases in both maximal velocity and affinity of the transporter.

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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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Hath1 Inhibits Proliferation of Colon Cancer Cells Probably Through Up-regulating Expression of Muc2 and p27 and Down-regulating Expression of Cyclin D1

  • Zhu, Dai-Hua;Niu, Bai-Lin;Du, Hui-Min;Ren, Ke;Sun, Jian-Ming;Gong, Jian-Ping
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.12
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    • pp.6349-6355
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    • 2012
  • Previous studies showed that Math1 homologous to human Hath1 can cause mouse goblet cells to differentiate. In this context it is important that the majority of colon cancers have few goblet cells. In the present study, the potential role of Hath1 in colon carcinogenesis was investigated. Sections of paraffin-embedded tissues were used to investigate the goblet cell population of normal colon mucosa, mucosa adjacent colon cancer and colon cancer samples from 48 patients. Hath1 and Muc2 expression in these samples were tested by immunohistochemistry, quantitative real-time reverse transcription -PCR and Western blotting. After the recombinant plasmid, pcDNA3.1(+)-Hath1 had been transfected into HT29 colon cancer cells, three clones were selected randomly to test the levels of Hath1 mRNA, Muc2 mRNA, Hath1, Muc2, cyclin D1 and p27 by quantitative real-time reverse transcription-PCR and Western blotting. Moreover, the proliferative ability of HT29 cells introduced with Hath1 was assessed by means of colony formation assay and xenografting. Expression of Hath1, Muc2, cyclin D1 and p27 in the xenograft tumors was also detected by Western blotting. No goblet cells were to be found in colon cancer and levels of Hath1 mRNA and Hath1, Muc2 mRNA and Muc2 were significantly down-regulated. Hath1 could decrease cyclin D1, increase p27 and Muc2 in HT29 cells and inhibit their proliferation. Hath1 may be an anti-oncogene in colon carcinogenesis.

Apoptotic Effects of Eugenol-loaded Nanoemulsions in Human Colon and Liver Cancer Cell Lines

  • Majeed, Hamid;Antoniou, John;Fang, Zhong
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.21
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    • pp.9159-9164
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    • 2014
  • Background: In this study eugenol (EU) loaded nanoemulsions (NEs) emulsified with modified starch were prepared and their apoptotic potential against liver and colon cancer cells was examined in comparison with bulk EU. Materials and Methods: We prepared stable EU loaded NEs whcih were characterized by dynamic light scattering, centrifugation and gas chromatography. Furthermore, cell viability was determined using MTT assay, and apoptosis and cell cycle analysess by flow cytometry. Results: HB8065 (liver) and HTB37 (colon) cells when treated with EU:CA NEs demonstrated greater apoptotic cells percentages as evidenced by microscopic images and flow cytometric evaluations. It was observed that EU and EU:CA NE induced apoptosis in both cell lines via reactive oxygen species (ROS) generation. Conclusions: The present study demonstrated that ROS plays a critical role in EU and EU:CA NE induced apoptosis in HB8065 and HTB37 cells. This is the first report on the antiproliferative mechanisms of EU loaded NE.

Growth Inhibition and G2/M Phase Cell Cycle Arrest by 3,4,5-Trimethoxy-4'-bromo-cis-stilbene in Human Colon Cancer Cells

  • Heo, Yeon-Hoi;Min, Hye-Young;Kim, Sang-Hee;Lee, Sang-Kook
    • Biomolecules & Therapeutics
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    • v.15 no.2
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    • pp.95-101
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    • 2007
  • Resveratrol (3,5,4’-trihydroxy-trans-stilbene), a naturally occurring phytoallexin abundant in grapes and several plants, has been shown to be active in inhibiting proliferation and inducing apoptosis in several human cancer cell lines. On the line of the biological activity of resveratrol, a variety of resveratrol analogs were synthesized and evaluated for their growth inhibitory effects against several human cancer cell lines. In the present study, we found that one of the resveratrol analogs, 3,4,5-trimethoxy-4’-bromo-cis-stilbene, markedly suppressed human colon cancer cell proliferation (EC$_{50}$ = 0.01 ${\mu}$g/ml), and the inhibitory activity was superior to its corresponding trans-isomer (EC$_{50}$ = 1.6 ${\mu}$g/ml) and resveratrol (EC$_{50}$ = 18.7 ${\mu}$g/ml). Prompted by the strong growth inhibitory activity in cultured human colon cancer cells (Col2), we investigated its mechanism of action. 3,4,5-Trimethoxy-4’-bromo-cis-stilbene induced arrest of cell cycle progression at G2/M phase and increased at sub-G1 phase DNA contents of the cell cycle in a time- and dose-dependent manner. Colony formation was also inhibited in a dose-dependent manner, indicating the inhibitory activity of the compound on cell proliferation. Moreover, the morphological changes and condensation of the cellular DNA by the treatment of the compound were well correlated with the induction of apoptosis. These data suggest the potential of 3,4,5-trimethoxy-4’-bromo-cis-stilbene might serve as a cancer chemotherapeutic or chemopreventive agent by virtue of arresting the cell cycle and inducing apoptosis for the human colon cancer cells.

Cytotoxic Effect of Inonotus obliquus Composition in HCT-15 Human Colon Cancer Cells and AGS Gastric Cancer Cells (대장암 세포암종 HCT-15 세포 및 위암 세포암종 AGS 세포에서 차가버섯 조성물에 의한 세포생육 억제 효과)

  • 차재영;전병삼;문재철;유지현;조영수
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.33 no.4
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    • pp.633-640
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    • 2004
  • This study was performed to investigate the cytotoxic effect of the water-extract from Chaga mushroom (Inonotus obliquus) compositions containing powdered green tea in HCT-15 human colon carcinoma, AGS human gastric carcinoma and NIH3T3 mouse normal fibroblast cells using viable cell count and MTT assay. The water-extract from Chaga mushroom compositions induced inhibitory effects on proliferation of HCT-15 and AGS cells in the MTT assay and viable cell count. However, mouse normal NIH3T3 cells were exhibited 80% survival under the same condition. Chaga mushroom compositions showed highly antiproliferative effect in human cancer cell line HCT-15 and AGS, but not in mouse normal cell line NIH3T3. These results suggest that Chaga mushroom (Inonotus obliquus) compositions containing powdered green tea are the candidate for chemoprevention in colon and gastric cancer.

Effects on Hot Water Extract of Schizandra chinensis on Colon Cancer (오미자 열수추출물의 대장암세포 증식억제 효과)

  • Ryu, Min-Ju;Chung, Ha-Sook
    • Food Engineering Progress
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    • v.15 no.1
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    • pp.64-69
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    • 2011
  • The anticancer activity of Schizandra chinensis Baillon was investigated for the development of functional food resources. The antiproliferative activity of hot water extracts of Schizandra chinensis Baillon in human colon cancer cell line (HT-29) were identified using cell viability, morphology study, cell cycle and RT-PCR analyses. HT-29 cells were cultured in several concentrations (0, 1.0, 2.0, 4.0 mg/mL) of water extracts of Schizandra chinensis Baillon. In our study, colon cancer cell growth could be inhibited by hot water extracts of Schizandra chinensis Baillon in a dose-dependent manners. It was associated with morphological changes and apoptotic cell death with cell shrinking, chromatin condensation, apoptotic bodies and cell cycle analysis. These results suggest that Schizandra chinensis Baillon may inhibit the growth of human colon cancer cells by various apoptosis-aiding activities as well as apoptosis itself.

The Antiproliferative Effects of Bile Acids and Their Derivatives on HT-29 Human Colon Cancer Cells

  • Park, Sang-Eun;Yee, Su-Bog;Choi, Hye-Joung;Chung, Sang-Woon;Park, Hwa-Sun;Yoo, Young-Hyun;Kim, Nam-Deuk
    • Proceedings of the PSK Conference
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    • 2003.04a
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    • pp.229.1-229.1
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    • 2003
  • The anti proliferative effects of bile acids and their derivatives on HT -29 human colon cancer cells were investigated. Ursodeoxycholic acid (UDCA) and its synthetic derivatives, HS-1030 and HS-1183, and chenodeoxycholic acid (CDCA) and its synthetic derivatives, HS-1199 and HS-1200 were employed for this study. General evaluations focusing on cell cycle were conducted in HT -29 human colon adenocarcinoma cell line (p53 mutant type). (omitted)

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Effect of Root Extracts of Medicinal Herb Glycyrrhiza glabra on HSP90 Gene Expression and Apoptosis in the HT-29 Colon Cancer Cell Line

  • Nourazarian, Seyed Manuchehr;Nourazarian, Alireza;Majidinia, Maryam;Roshaniasl, Elmira
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.18
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    • pp.8563-8566
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    • 2016
  • Colorectal cancer is one of the most common lethal cancer types worldwide. In recent years, widespread and large-scale studies have been done on medicinal plants for anti-cancer effects, including Glycyrrhiza glabra. The aim of this study was to evaluate the effects of an ethanol extract Glycyrrhiza glabra on the expression of HSP90, growth and apoptosis in the HT-29 colon cancer cell line. HT-29 cells were treated with different concentrations of extract (50,100,150, and $200{\mu}g/ml$). For evaluation of cell proliferation and apoptosis, we used MTT assay and flow cytometry technique, respectively. RT-PCR was also carried out to evaluate the expression levels of HSP90 genes. Results showed that Glycyrrhiza glabra inhibited proliferation of the HT-29 cell line at a concentration of $200{\mu}g/ml$ and this was confirmed by the highest rate of cell death as measured by trypan blue and MTT assays. RT-PCR results showed down-regulation of HSP90 gene expression which implied an ability of Glycyrrhiza glabra to induce apoptosis in HT-29 cells and confirmed its anticancer property. Further studies are required to evaluate effects of the extract on other genes and also it is necessary to make an extensive in vivo biological evaluation and subsequently proceed with clinical evaluations.

Heptaphylline Induces Apoptosis in Human Colon Adenocarcinoma Cells through Bid and Akt/NF-κB (p65) Pathways

  • Boonyarat, Chantana;Yenjai, Chavi;Vajragupta, Opa;Waiwut, Pornthip
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.23
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    • pp.10483-10487
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    • 2015
  • Heptaphylline derivatives are carbazoles in Clausena harmandiana, a medicinal plant that is utilized for headache, stomach ache, and other treatments of illness. The present study examined the effects of heptaphylline and 7-methoxyheptaphylline on apoptosis of human colon adenocarcinoma cells (HT-29 cell line). Quantification of cell viability was performed using cell proliferation assay (MTT assay) and of protein expression through immunoblotting. The results showed that only heptaphylline, but not 7-methoxyheptaphylline, significantly significantly activated cleaved of caspase-3 and poly (ADP-ribose) polymerase (PARP-1) which resulted in HT-29 cell death. We found that heptaphylline activated BH3 interacting-domain death agonist (Bid) and Bak, proapoptotic proteins. In contrast, it suppressed X-linked inhibitor-of-apoptosis protein (XIAP), Bcl-xL and survivin, inhibitors of apoptosis. In addition, heptaphylline inhibited activation of NF-${\kappa}B$/p65 (rel), a regulator of apoptotic regulating proteins by suppressing the activation of Akt and $IKK{\alpha}$, upstream regulators of p65. The findings suggested that heptaphylline induces apoptosis in human colon adenocarcinoma cells.