• 제목/요약/키워드: human colon cancer cells

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Anti-Proliferative Effect of Polysaccharides from Salicornia herbacea on Induction of G2/M Arrest and Apoptosis in Human Colon Cancer Cells

  • Ryu, Deok-Seon;Kim, Seon-Hee;Lee, Dong-Seok
    • Journal of Microbiology and Biotechnology
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    • v.19 no.11
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    • pp.1482-1489
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    • 2009
  • In this study, we investigated the anti-proliferative effect of polysaccharides from Salicornia herbacea on HT-29 human colon cancer cells. Crude polysaccharides from S. herbacea (CS) were prepared by extraction with hot steam water, and fine polysaccharides from S. herbacea (PS) were obtained through further size exclusion chromatography. The anti-proliferative effect of CS and PS were measured using the MTS assay, apoptosis analysis, cell cycle analysis, and RT-PCR. HT-29 cells were treated with CS or PS at different dosages (0.5, 1, 2, 4 mg $ml^{-1}$) for 24 or 48 h. CS and PS inhibited proliferation and stimulated apoptosis of cells in a dose-dependent manner. Flow cytometric analysis after Annexin V-FITC and PI staining revealed that treatment with CS or PS increased total apoptotic death of cells to 24.99% or 91.59%, respectively, in comparison with the control (13.51 %). PS increased early apoptotic death substantially - up to 12 times more than the control. Treatment with CS or PS resulted in a concentration-dependent increase of the G2/M cell population of the cell cycle as determined by flow cytometry. G2/M arrest was induced significantly with the highest concentration (4 mg $ml^{-1}$) of PS. RT-PCR was performed to study the correlation between G2/M arrest and transcription of cell cycle control genes. The anti-proliferative activity of CS and PS was accompanied by inhibition of cyclin B1, and Cdc 2 mRNA. Moreover, both CS and PS induced expression of the p53 tumor suppressor gene and the Cdk inhibitor p21. These results suggest that polysaccharides from S. herbacea have anti-cancer activity in human colon cancer cells.

Monitoring the Change of Protein Expression in Human Colon Cancer Cell SNU-81 treated with the Water-Extract of Coptis japonica (황련 열수추출물을 처치한 인간 대장암 세포 SNU-81에서의 단백질 발현 변화)

  • Yoo, Tae-Mo;Kim, Byung-Soo;Yoo, Byong-Chul;Yoo, Hwa-Seung
    • Journal of Pharmacopuncture
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    • v.12 no.1
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    • pp.5-12
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    • 2009
  • Background : Anticancer effects of herbal medicine have been reported in various types of cancer, but the systematic approaches to explain molecular mechanism(s) are not established yet. Objective : To find the anticancer-effect and mechanism(s) of Water Extract of Coptis japonica (WECJ) colon cancer cell (SNU-81). Methods : We first selected 11 herbals, and anti-cancer effects of water-extracts from those herbals have been tested in human colon cancer cell line, SNU-81. Among the tested herbals, the WECJ significantly reduced proliferation of SNU-81. To establish a basis of understanding for anti-cancer mechanism, whole proteins have been obtained from SNU-81 harvested at 48 and 96 hrs after the treatment of WECJ, protein expression has been profiled by 2DE-based proteomic approach. Results : Various changes of the protein expression have been monitored, and most frequent dysregulation was found in the molecular chaperons including heat shock protein 90-alpha (Hsp90-alpha), 14-3-3 protein epsilon, T-complex protein 1 subunit alpha, protein disulfide-isomerase A3, and calreticulin. Interestingly, proliferation-associated protein 2G4 has been up-regulated, and it suggests the possible effect of Coptis japonica on ErbB3-regulated signal transduction pathway and growth control of human colon cancer cells. Conclusion : Based upon the present findings, the further study will focus on monitoring various cancer survival factors after artificial regulation of the proteins identified, and it would be the basis for the understanding of the Coptis japonica anti-cancer effect(s) at the molecular level.

Anticancer Effect of Doenjang in in vitro Sulforhodamine B (SRB) Assay (된장의 in vitro Sulforhodamine B (SRB) Assay에 의한 암세포 증식 억제 효과)

  • 이숙희;임선영;박건영
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.28 no.1
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    • pp.240-245
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    • 1999
  • Growth inhibitory effect of doenjang(Korean soypaste) methanol extracts in SRB assay using AGS human gastric adenocarcinoma cell, Hep 3B human hepatocellular carcinoma cell and HT 29 human colon cancer cell was studied. The treatment of doenjang methanol extracts(2mg/assay) to the AGS, Hep 3B and HT 29 cancer cells inhibited the growth of the cancer cells by 55%, 60%, and 71%, respectively. Doenjang methanol extracts exhibited the highest inhibitory effect among other soybean fermented foods and original materials in the SRB assay. In addition, to separate active compounds of doenjang methanol extracts, we fractionated the doenjang with hexane, methanol, dichloromethane, ethylacetate and butanol. Growth inhibitory effect on the AGS, Hep 3B, HT 29 and MG 63 cancer cells was the highest in the fractions of dichloromethane and ethylacetate among other solvent fractions of the doenjang. These results showed that some compounds contained in the fractions of dichloromethane and ethylacetate might play a role on the anticanceric effect of doenjang.

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Active Immunization Study of Colon Cancer Derived 1-8D Peptide in HHD Mice

  • Jung, Hun-Soon;Ahn, In-Sook;Do, Hyung-Ki;Lemonnier, Francois A.;Song, Kuk-Hyun;Do, Myoung-Sool
    • IMMUNE NETWORK
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    • v.5 no.3
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    • pp.157-162
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    • 2005
  • Background: 1-8D gene is a member of human 1-8 interferon inducible gene family and was shown to be overexpressed in fresh colon cancer tissues. Three peptides 1-6, 3-5 and 3-7 derived from human 1-8D gene were shown to have immunogenicity against colon cancer. Methods: To study tumor immunotherapy, of three peptides we established an active immunization model using HHD mice. $D^{b-/-}{\times}{\beta}2$ microglobulin $({\beta}2m)$ null mice transgenic for a chimeric HLA-$A2.1/D^{b-}\;{\beta}2m$ single chain (HHD mice) were challenged with B16/HHD/1-8D tumor cells and were immunized with irradiated peptide-loaded RMA- S/HHD/B7.1 transfectants. In therapy model tumor growth was retarded in HHD mice that were injected with 3-5 peptide-loaded RMA-S/HHD/B7.1. In survival test vaccination with 1-8D-derived peptide protects HHD mice from tumor progression after tumor challenge. Results: These studies show that peptide 3-5 derived from 1-8D gene can be the most effective candidate for the vaccine of immunotherapy against colon cancer and highlight 1-8D gene as putative colon carcinoma associated antigens. Conclusion: We demonstrated that RMA-S/HHD/ B7.1 loaded with 1-8D peptides, especially 3-5, immunization generates potent antitumor immunity against tumor cells in HHD mice and designed active immunization as proper immunotherapeutic protocols.

Methanol Extract of Flacourtia indica Aerial Parts Induces Apoptosis via Generation of ROS and Activation of Caspases in Human Colon Cancer HCT116 Cells

  • Park, Ki-Woong;Kundu, Juthika;Chae, In Gyeong;Bachar, Sitesh Chandra;Bae, Jung-Woo;Chun, Kyung-Soo
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.17
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    • pp.7291-7296
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    • 2014
  • Different plant parts of Flacourtia indica have long been used in Ayurvedic medicine. Previous studies have demonstrated that the methanolic extract of F. indica possess anti-inflammatory properties. The present study was aimed at investigating the anticancer effects of methanol extract of Flacourtia indica (FIM) aerial parts in human colon cancer (HCT116) cells. Treatment of cells with FIM at a concentration of $500{\mu}g/ml$ for 24 hours significantly reduced cell viability and induced apoptosis, which was associated with the increased cytoplasmic expression of cytochrome c, activation of caspase-3, and the cleavage of poly-(ADP-ribose) polymerase. Incubation with FIM also inhibited the levels of Bcl-2, Bcl-xl and survivin, which are the markers of cell proliferation, whereas the expression of Bax remained unchanged. Treatment with FIM led to the generation of reactive oxygen species (ROS) in a concentration-dependent manner. Pharmacological inhibition of ROS generation by pretreatment of cells with N-acetyl cysteine abrogated FIM-induced apoptosis in HCT116 cells. Thus, these results demonstrate that FIM has anti-proliferative and pro-apoptotic effects in HCT116 cells and the effects are, at least in part, due to the ROS dependent activation of caspases.

Anti-Proliferative Activity of Nodosin, a Diterpenoid from Isodon serra, via Regulation of Wnt/β-Catenin Signaling Pathways in Human Colon Cancer Cells

  • Bae, Eun Seo;Kim, Young-Mi;Kim, Dong-Hwa;Byun, Woong Sub;Park, Hyen Joo;Chin, Young-Won;Lee, Sang Kook
    • Biomolecules & Therapeutics
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    • v.28 no.5
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    • pp.465-472
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    • 2020
  • Colorectal cancer (CRC) is one of the most malignant type of cancers and its incidence is steadily increasing, due to life style factors that include western diet. Abnormal activation of canonical Wnt/β-catenin signaling pathway plays an important role in colorectal carcinogenesis. Therefore, targeting Wnt/β-catenin signaling has been considered a crucial strategy in the discovery of small molecules for CRC. In the present study, we found that Nodosin, an ent-kaurene diterpenoid isolated from Isodon serra, effectively inhibits the proliferation of human colon cancer HCT116 cells. Mechanistically, Nodosin effectively inhibited the overactivated transcriptional activity of β-catenin/T-cell factor (TCF) determined by Wnt/β-catenin reporter gene assay in HEK293 and HCT116 cells. The expression of Wnt/β-catenin target genes such as Axin2, cyclin D1, and survivin were also suppressed by Nodosin in HCT116 cells. Further study revealed that a longer exposure of Nodosin induced the G2/M phase cell cycle arrest and subsequently apoptosis in HCT116 cells. These findings suggest that the anti-proliferative activity of Nodosin in colorectal cancer cells might in part be associated with the regulation of Wnt/β-catenin signaling pathway.

The p53-p21Cip1/WAF1 Pathway Is Necessary for Cellular Senescence Induced by the Inhibition of Protein Kinase CKII in Human Colon Cancer Cells

  • Kang, Ji-Young;Kim, Jin Joo;Jang, Seok Young;Bae, Young-Seuk
    • Molecules and Cells
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    • v.28 no.5
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    • pp.489-494
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    • 2009
  • We have previously shown that the down-regulation of protein kinase CKII activity is tightly associated with cellular senescence of human fibroblast IMR-90 cells. Here, we examined the roles of p53 and $p21^{Cip1/WAF1}$ in senescence development induced by CKII inhibition using wild-type, isogenic p53-/- and isogenic p21-/- HCT116 human colon cancer cell lines. A senescent marker appeared after staining for senescence-associated ${\beta}$-galactosidase activity in wild-type HCT116 cells treated with CKII inhibitor or $CKII{\alpha}$ siRNA, but this response was almost abolished in p53- or $p21^{Cip1/WAF1}$-null cells. Increased cellular levels of p53 and $p21^{Cip1/WAF1}$ protein occurred with the inhibition of CKII. CKII inhibition upregulated p53 and $p21^{Cip1/WAF1}$ expression at post-transcriptional level and transcription level, respectively. RB phosphorylation significantly decreased in cells treated with CKII inhibitor. Taken together, this study shows that the activation of the $p53-p21^{Cip1/WAF1}$ pathway acts as a major mediator of cellular senescence induced by CKII inhibition.

Role of p53-dependent PI3K in Radioresistance of Colon Cancer Cells (대장암 세포의 방사선저항성에 대한 p53의존성 PI3K의 역할)

  • Lee, Heui-Kwan;Kim, Jong-Suk;Kwon, Hyoung-Cheol
    • Journal of Food Hygiene and Safety
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    • v.25 no.3
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    • pp.258-262
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    • 2010
  • Radiotherapy is one of the major therapies for cancer treatment. p53 acts as a central mediator of the cellular response to stressful stimuli, such as radiation. Recently it has been known that activation of the phosphatidylinositol-3-kinase (PI3K) pathway is associated with radioresistance. In this study, we investigated whether X-irradiation up-regulates PI3K in a p53-dependent manner in human colon cancer cells. In order to study this phenomenon, we have treated p53-wild type and p53-mutant type HCT116 cells with X-ray. Treatment of wild type HCT116 cells with 8 Gy resulted in a marked increase in PI3K (p85), which paralleled an increase in PTEN, a counterpart of PI3K. However, these effects of X-rays in the p53-mutant cells were not observed. These results suggest that the X-irradiation-induced up-regulation of PI3K/PTEN pathway is p53-dependent.

Effects of Korean and Chinese Crataegi Fructrus on the Antioxidant Activity and Antiproliferation of Cancer Cells (한국산과 중국산 산사의 항산화 활성 및 암세포 증식 억제효과)

  • Park, Yonghyun;Lee, Hyun-Joo;Lee, Jae-Joon
    • The Korean Journal of Community Living Science
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    • v.26 no.1
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    • pp.103-113
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    • 2015
  • This study examines the effects of Korean Crataegi fructrus(KCF) and Chinese Crataegi fructrus(CCF) on the antioxidative activity and antiproliferation of human cancer cells(HCT-116 human colon, Hep G2 human liver, and A549 human lung cancer cells). The total polyphenol and flavonoid contents, and antioxidative index of the Crataegi fructrus ethanol extracts were significantly higher in KCF than in CCF. The DPPH radical-scavenging activity of the KCF ethanol extract was 82.26%(1000 ppm), and that of the CCF ethanol extract was 77.64%. Antiproliferation effects of 80% ethanol extracts of KCF and CCF on human cancer cells(HCT-116, Hep G2 and A549) increased in a dose-dependent manner. Inhibitory effects of KFC on HCT-116 and A549 cells were greater than those of CCF. The results suggest that ethanol extracts of Crataegi fructrus have antioxidative and hyperplasia inhibition effects on human cancer cells.

Effects of Dietary Fiber from Mozuku (Cladosiphon novae-caledoniae kylin) Residue on Antioxidant Activity and Anticancer in HT-29 Human Colon Cancer Cells according to Extraction Condition (추출법에 따른 큰실말(Cladosiphon novae-caledoniae kylin) 부산물 식이섬유의 항산화 효과 및 HT-29 대장암 세포에 대한 항암 작용)

  • Jeong, Haeng Soon;Lee, Jae-Hwa
    • Applied Chemistry for Engineering
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    • v.25 no.4
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    • pp.363-367
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    • 2014
  • In this study, the dietary fiber contents of Mozuku (Cladosiphon novae-caledoniae kylin) residue and the extraction condition (HCl, $H_2SO_4$, NaOH, $Na_2CO_3$, $Na_2EDTA$) of the dietary fiber was investigated. We examined that the contents of the total polyphenols and flavonoids in the dietary fiber from Mozuku residue, and the potent anti-cancer effect was also tested through the growth inhibition in human colon cancer cells (HT-29) in vitro. It was effective to extract soluble dietary fiber with 1.5% $Na_2EDTA$ and 0.05 N HCl in Mozuku residue. The extraction time and temperature affected the yields of soluble dietary fiber. The contents of the total polyphenols and flavonoids in the dietary fiber from Mozuku residue were the highest in 1% NaOH extract (Total polyphenols $34.4{\pm}0.055$ mg gallic acid/g dry basis, total flavonoids $34.7{\pm}0.023$ mg naringin/g extract dry basis). In DPPH radical scavenging activity, 1% NaOH extract showed the most potent antioxidant activity. In the result of viability in human colon cancer cells, growth inhibition was observed in D.W., 0.05 N HCl, and 0.5% $Na_2CO_3$ extracts in a dose-dependent manner. These results demonstrated that soluble dietary fiber from Mozuku residue significant antioxidant activity and anticancer in human colon cancer.