• Title/Summary/Keyword: HT-29 human

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Antiproliferative Effect and Apoptotic Induction of Bauhinia forficata Extract in Human Cancer Cells. (Bauhinia forficata 추출물의 인체 암세포에 대한 성장억제 및 세포사멸 유도 활성)

  • 임혜영;이철훈
    • Microbiology and Biotechnology Letters
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    • v.32 no.1
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    • pp.67-71
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    • 2004
  • Pata de Vaca (Bauhinia forficata) Is a tree which grows naturally in the rainforests and tropical parts of Peru and Brazil, as well as tropical zones of Asia, eastern Paraguay and northeastern Argentina. The active fraction (Pata-50) of the 70% ethanol extract from Pata de Vaca was sequentially fractionated by HP-20 Diaion column chromatography and C-18 column chromatography, and its characteristics were investigated. The growth of all cancer cells tested except for MCF-7 was Inhibited in a concentration-dependent manner by Pata-50. Its $IC_{50}$ values were estimated to be 40.4 $\mu\textrm{g}$/$m\ell$ on AGS, 51.3 $\mu\textrm{g}$/$m\ell$ on HT-29, 52.1$\mu\textrm{g}$/$m\ell$ on HepG2, 65.2$\mu\textrm{g}$/$m\ell$ on A549, and 77.5$\mu\textrm{g}$/$m\ell$ on HeLa cells. A flow cytometric analysis of HepG2 cells revealed induction of apoptosis, but cell cycle regulation was not affected. The HepG2 cell population of apoptosis region increased In a concentration-dependent manner by Pata-50.

Growth Inhibitory and Quinone Reductase Activity Stimulating Effects of Internal Organs of Aplysia kurodai Fractions on Cancer Cell Lines In vitro (군소내장 분획물의 in vitro에서의 암세포 성장억제 및 quinone reductase 유도 활성 증가 효과)

  • Shin, Mi-Ok
    • Journal of Life Science
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    • v.20 no.6
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    • pp.877-884
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    • 2010
  • We investigated the growth inhibitory effect of internal organs of Aplysia kurodai (AK) on proliferation in cancer cell lines in vitro. The internal organs of AK were extracted with methanol (AKM), which were then further fractionated into four subfractions by using solvent partition method, resulting in hexane (AKMH), methanol (AKMM), butanol (AKMB), and aqueous (AKMA) soluble fractions. We determined the cytotoxic effect of these four fractions in four kinds of cancer cell lines - HepG2, MCF-7, HT29 and B16-F10 - by MTT assay. Among the four subfractions of AKM, AKMM showed the strongest cytotoxic effects on all cancer cell lines which were used. Morphological changes such as membrane shrinking and blebbing of cells were also observed in AKMM treatment in HepG2 cells. In addition, we also observed quinone reductase (QR) induced effect in the methanol layer (AKMM) of HepG2 cells. AKMM showed the highest induction activity of quinone reductase on HepG2 cells among the partition layers. The QR induced effect of AKMM was determined to be 2.4 at $100\;{\mu}g/ml$ level with a control value of 1.0. Although further studies are needed, the present work suggests that internal organs of Aplysia kurodai (AK) may be a chemopreventive agent for the treatment of human cells.

Antioxidant and Anticancer Activities of Extracts from Styela plicata (주름 미더덕(Styela plicata) 추출물의 항산화력 및 항암활성)

  • Kim, Jin-Ju;Kim, Sun-Jung;Kim, Sun-Hee;Park, Hae-Ryong;Lee, Seung-Cheol
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.7
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    • pp.937-941
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    • 2005
  • The antioxidant and anticancer activities of Styela plicata extracts were evaluated. When extracts were prepared with fresh Styela plicata (FR), extraction yield was in the order of methanol > ethanol = acetone > water among treated solvents. However, the extraction order was methanol > water > ethanol > acetate in freeze dried Styela plicata (FD). Radical scavenging activity was the highest in acetone extracts $(37.39\%)$ from FR, while in ethanol extracts $(78.40\%)$ from FD. Reducing power of FR was the greatest in methanol extracts (1.076), and that of FD in ethanol extracts (1.360). The acetone extracts from FD showed significant anticancer activity when revealed with human colon cancer cell line HT-29. These results indicated that extraction yields and properties of extracts from Styela plicata were variable depending on solvent and/or physicochemical state, and appropriate extraction process could provide some valuable bioactive materials from Styela plicata.

Biological Activities of Crude Polysaccharides and Crude Saponins from Salicornia herbacea (함초 조다당체 및 조사포닌의 생리활성)

  • Jung, Sung-Hee;Park, Kyung-Uk;Kim, Jae-Yong;Park, Chae-Kyu;Choi, Kap-Seong;Seo, Kwon-Il
    • Food Science and Preservation
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    • v.16 no.1
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    • pp.109-114
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    • 2009
  • To develop Salicornia herbacea as a functional food material, crude polysaccharides and saponins were isolated from 80% ethanolic extracts of Salicornia herbacea using Diaion HP-20 column chromatography, and biological activities including anti-oxidation, anti-proliferation of cancer cells, and immune activities were investigated. The hydrogen-donating properties of crude polysaccharides and saponins were above 20% at $500{\mu}g/mL$ concentration. The reducing power of fractions increased in a dose-dependent manner. Crude polysaccharides and saponins at $500{\mu}g/mL$ inhibited more than 20% and 50% of proliferation by PC-3 and HT-29 human cancer cells, respectively. The anti-oxidative and anti-cancer properties of crude saponins were higher than those of crude polysaccharides. Also, proliferation in spleen cells and (nitric oxide) NO production in RAW264.7 macrophages treated with crude polysaccharides increased in a dose-dependent manner compared with the untreated control cells. However, crude saponins at concentration greater than $500{\mu}g/mL$ resulted in decreases in immune activities. Immune activities of crude polysaccharide were greater than those of crude saponins. These results indicate that Salicornia herbacea may be used as functional food materials.

Ginseng berry polysaccharides on inflammation-associated colon cancer: inhibiting T-cell differentiation, promoting apoptosis, and enhancing the effects of 5-fluorouracil

  • Wang, Chong-Zhi;Hou, Lifei;Wan, Jin-Yi;Yao, Haiqiang;Yuan, Jinbin;Zeng, Jinxiang;Park, Chan Woong;Kim, Su Hwan;Seo, Dae Bang;Shin, Kwang-Soon;Zhang, Chun-Feng;Chen, Lina;Zhang, Qi-Hui;Liu, Zhi;Sava-Segal, Clara;Yuan, Chun-Su
    • Journal of Ginseng Research
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    • v.44 no.2
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    • pp.282-290
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    • 2020
  • Background: Ginseng is a commonly used herbal medicine in treating various medical conditions. Chronic gut inflammation is a recognized factor for the development of colorectal cancer (CRC). In this project, Asian ginseng berry polysaccharide preparations were used to assess their effects on CRC and related immune regulation mechanisms. Methods: Ginseng berry polysaccharide extract (GBPE) and purified ginseng berry polysaccharide portion (GBPP) were used to evaluate their activities on human HCT-116 and HT-29 CRC cell proliferation. Interleukin-8 secretion analysis was performed on HT-29 cells. Naive CD4 cell isolation and T-helper cell differentiation were performed and determined using flow cytometry for Th1 and Treg in addition to cell cycle and apoptotic investigation. Results: GBPE and GBPP significantly inhibited interleukin-8 secretion and cancer cell proliferation, inhibited CD4+IFN-γ+ cell (Th1) differentiation, and decreased CD4+FoxP3+ cell (Treg) differentiation. Compared to the GBPE, GBPP showed more potent antiinflammatory activities on the malignant cells. This is consistent with the observation that GBPP can also inhibit Th1-cell differentiation better, suggesting that it has an important role in antiinflammation, whereas Treg cells hinder the body's immune response against malignancies. Supported by cell cycle and apoptosis data, GBPE and GBPP, at various degrees, remarkably enhanced the anticancer activities of 5-fluorouracil. Conclusion: Data from this project suggested that Asian ginseng berry potentially has clinical utility in managing enteric inflammation and suppressing CRC through immunomodulation mechanisms.

Detection of Human Taurine Transporter and Production of Monoclonal Antibody

  • An, Hye-Suk;Han, Hee-Chang;Lee, Sun-Min;Park, Taesun;Park, Kun-Koo;Kim, Ha-Won
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 2001.11a
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    • pp.102-102
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    • 2001
  • Taurine (2-ethaneaminosulfonic acid) is one of the major intracellular ${\beta}$ -amino acids in mammals and is required for a number of biological processes including membrane stabilization, osmoregulation, antioxidation, detoxification, modulation of calcium flux and neurornodulation. The taurine transporter (TAUT) which contains 12 hydrophobic membrane-spanning domains has been cloned from dog kidney, rat brain, mouse brain, human thyroid, placenta and retina. In this study, The TAUT cDNA from the human intestinal epithelial cell, HT-29 was cloned and sequenced. Reverse-transcription polymerase chain reaction (RT-PCR) was performed to amplify partial cDNA encoding human intestinal TAUT. The coding region of the PCR product was 732 bp long. The primers were designed to encode highly conserved amino acid sequences near the transmembrane domains III (IPYFIFLF) and Ⅵ (KYKYNSYR) both in human and mouse. The TAUT cDNA amplified was ligated into the pGEX 4T-1 expression vector. The resulting sequence of human intestinal TAUT cDNA (Accession number of NCBI Genebank is AF346763) was identical to the sequences of the TAUTs previously determined in the human placenta and retina except 3 base pairs from that of the reported human thyroid. TAUT specific antibodies were generated to use them as biological tools in the studies of the biological role of TAUT. Peptides of 149-162 amino acid residue (14 amino acids) of the TAUT were synthesized. The synthetic peptide used in this study was LFQSFQKELPWAHC. This region was chosen not only to avoid putative glycosylation sites but also to exclude regions of known homology with GABA transporters in the extracellular hydrophilic domains. The synthetic peptide, TAUT-1 was conjugated with carrier protein, kehole lympet hemocyanin (KLH) to use as an antigen. When used for immunization on a rabbit to produce polyclonal antiserum, the conjugates elicited high -titered specific anti-TAUT-1 antibodies, which reacted well with the ovalbumin (OVA) conjugated peptides in ELISA. The KLH-conjugated peptide was also used as immunizing antigen in BALB/c mice to produce TAUT specific monoclonal antibodies. From the culture supernatant of the hybridoma, the specificity of anti-TAUT-1 monoclonal antibodies was confirmed by ELISA. Further applications of more tools in TAUT expression analysis will be performed such as western blotting and flow cytometry.

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Cytotoxic and Antimutagenic Stilbenes from Seeds of Paeonia lactiflora

  • Kim, Hyo-Jin;Chang, Eun-Ju;Bae, Song-Ja;Shim, Sun-Mi;Park, Heui-Dong;Rhee, Chang-Ho;Park, Jun-Hong;Choi, Sang-Won
    • Archives of Pharmacal Research
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    • v.25 no.3
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    • pp.293-299
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    • 2002
  • Cytotoxic and antimutagenic effects of a novel cis-$\varepsilon$-viniferin and five known stilbenes, transresveratrol, trans-$\varepsilon$-viniferin, gnetin H, suffruticosols A and B, isolated from the seeds of Paeonia lactiflora Pall. (Paeoniaceae) were determined against five different cancer cell lines, and mutagenicity of N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) in Salmonella typhimurium TA100, respectively. Six stilbenes showed cytotoxic activity in a dose-dependent manner, and especially did potent cytotoxic activity against C6 (mouse glioma) cancer cell with $IC_{50}$ values ranging from 8.2 to $20.5{\;}{\mu\textrm{g}}/ml$. trans-Resveratrol showed significant cytotoxic activity against HepG2 (liver hepatoma) and HT-29 (colon) human cancer cell lines with $IC_{50}$ values of 11.8 and 25.2 g/ml, respectively. In contrast, trans-$\varepsilon$-viniferin and cis--viniferin, and gnetin H exhibited marked cytotoxic activity against Hela (cervicse) and MCF-7 (breast) human cancer cell lines with $IC_{50}$ values of 20.4, 21.5, and $12.9{\;}{\mu\textrm{g}}/ml$, respectively. However, suffruticosol A and B had less cytotoxic effect against all cancer cells except C6. Meanwhile, six stilbenes exerted antimutagenic activity in a dose-dependent fashion. Of them, trans-resveratrol exhibited the strongest antimutagenic effect against MNNG with $IC_{50}$ value of $27.0{\;}{\mu\textrm{g}}/plate$, while other five resveratrol oligomers also did moderate antimutagenic activity with $IC_{50}$ values ranging from 31.7 to $35.2{\;}{\mu\textrm{g}}/plate$.

Physico-chemical Properties and In Vivo Anti-cancer Effects of Potato Kimchi Prepared by adding Hot Water Extracts of Potato (생감자의 열탕 추출물을 첨가한 감자 김치의 이화학적 특성 및 In Vivo에서의 항암 효과)

  • Chang, Sang-Keun;Kim, Hee-Joo
    • Journal of the East Asian Society of Dietary Life
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    • v.18 no.3
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    • pp.302-310
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    • 2008
  • In this study, potato kimchi was prepared by applying heat to raw potatoes, and then the physico-chemical properties and anti-cancer effects of the kimchi were analyzed. The texture results indicated the potato kimchi had very good hardness and springiness attributes. During th late storage period, total vitamin C content of the kimchi slowly increased. In addition, the potato kimchi had non-volatile organic acid changes that promoted early aging; however, after the complete aging period, it was comparatively similar to other types of kimchi. Using the methanol extracts of various kimchi samples, the potato kimchi(solid 100%) showed the highest anti-carcinogenic effects in terms of anti-tumor activity in tumor bearing Balb/c mice with sarcoma-180 cells. In addition, the effects of the methanol extracts on hepatic glutathione S-transferase content were $289.76\;{\mu}mol/mg$ protein/min, $250.97\;{\mu}mol/mg$ protein/min, $251.20\;{\mu}mol/mg$ protein/min, $219.53\;{\mu}mol/mg$ protein/min, $183.79\;{\mu}mol/mg$ protein/min, for control kimchi, mul kimchi, and two potato kimchis [(solid 100%) and(solid 60%+kimchi juice 40%)], respectively. The in vivo anti-cancer effects of the potato kimchi were investigated using AGS human gastric adenocarcionoma cells and HT-29 human colon adenocarcionoma cells. Overall, an MTT assay revealed that the methanol extract of the potato kimchi showed the highest anti-carcinogenic effects.

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Amelioration of DSS-Induced Acute Colitis in Mice by Recombinant Monomeric Human Interleukin-22

  • Suhyun Kim;Eun-Hye Hong;Cheol-Ki Lee;Yiseul Ryu;Hyunjin Jeong;Seungnyeong Heo;Joong-Jae Lee;Hyun-Jeong Ko
    • IMMUNE NETWORK
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    • v.22 no.3
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    • pp.26.1-26.18
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    • 2022
  • IL-22, a pleiotropic cytokine, is known to have a profound effect on the regeneration of damaged intestinal barriers. The tissue-protective properties of IL-22 are expected to be potentially exploited in the attenuation and treatment of colitis. However, because of the disease-promoting role of IL-22 in chronic inflammation, a comprehensive evaluation is required to translate IL-22 into the clinical domain. Here, we present the effective production of soluble human IL-22 in bacteria to prove whether recombinant IL-22 has the ability to ameliorate colitis and inflammation. IL-22 was expressed in the form of a biologically active monomer and non-functional oligomers. Monomeric IL-22 (mIL-22) was highly purified through a series of 3 separate chromatographic methods and an enzymatic reaction. We reveal that the resulting mIL-22 is correctly folded and is able to phosphorylate STAT3 in HT-29 cells. Subsequently, we demonstrate that mIL-22 enables the attenuation of dextran sodium sulfate-induced acute colitis in mice, as well as the suppression of pro-inflammatory cytokine production. Collectively, our results suggest that the recombinant mIL-22 is suitable to study the biological roles of endogenous IL-22 in immune responses and can be developed as a biological agent associated with inflammatory disorders.

Isolation and Biological Properties of Novel Cell Cycle Inhibitor, HY558, Isolated from Penicillium minioluteum F558

  • Lee, Chul-Hoon;Lim, Hae-Young;Kim, Min-Kyoung;Cho, Youl-Hee;Oh, Deok-Kun;Kim, Chang-Jin;Lim, Yoon-Gho
    • Journal of Microbiology and Biotechnology
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    • v.12 no.3
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    • pp.470-475
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    • 2002
  • In the course of screening for a novel cell cycle inhibitor, a potent Cdk 1 inhibitor, HY558, was found from the culture broth of Penicillium minioluteum F558 isolated from a soil sample. The molecular ion of HY558 was identified at m/z 329 (MH+) with a molecular formula of $C_20H_44ON_2$. HY558 exhibited selective antiproliferative effects on various human cancer cell lines. Its $IC_50$ values were estimated to be 0.29 mM on HepG2, 0.30 mM on HeLa, 0.30 mM on HL6O, 0.33 mM on HT-29, and 0.25 mM on AGS cells. Interestingly, Hy558 demonstrated no antiproliferative effect with normal lymphocytes used as the control, and a low level of inhibition on the proliferation of A549 cancer cells. A flow cytometric analysis of HepG2 cells revealed an appreciable arrest of cells at the G1 and G2/M phases of the cell cycle following treatment with Hy558. furthermore, DNA fragmentation due to apoptosis was observed in HeLa cells treated with 0.46 mM of HY558.