• Title/Summary/Keyword: HT-29 Cell

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Antioxidant and Anticancer Activities of Methanol Extracts Prepared from Different Parts of Jangseong Daebong Persimmon (Diospyros kaki cv. Hachiya) (장성 대봉감의 부위별 메탄올 추출물의 항산화 활성 및 항암 활성)

  • Jo, Young-Hong;Park, Ji-Won;Lee, Jeung-Min;Ahn, Gwang-Hwan;Park, Hae-Ryong;Lee, Seung-Cheol
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.4
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    • pp.500-505
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    • 2010
  • The antioxidant activity of methanol extracts from five different parts (flesh, peel, core, seed, calyx) of Jangseong Daebong persimmon (Diospyros kaki cv. Hachiya) were evaluated by determining total phenol content (TPC), DPPH radical scavenging activity (RSA), ABTS RSA, and reducing power (RP). The flesh extract gave the highest yield (92.93%) while the lowest yield was obtained from the seed (5.17%). The seed extract showed the highest total phenolic content ($76.47\pm0.009$ mg GAE/g extract), DPPH RSA ($IC_{50}=52.05\pm1.61\;{\mu}g/mL$), ABTS RSA ($IC_{50}=30.94\pm0.41\;{\mu}g/mL$) and RP ($IC_{50}=87.94\pm0.37\;{\mu}g/mL$). In addition, the calyx extract also showed high antioxidant activity. On the other hand, the core extract gave the lowest TPC and all antioxidant assays. In particular, HT-29 cells showed extensive cell death when treated with $500\;{\mu}g/mL$ of calyx extracts. Thus, these results suggest that methanolic extracts of Jangseong Daebong persimmon seed and calyx may serve as a potential source of natural antioxidant for food and nutraceutical application.

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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The Effect of (1S,2S,3E,7E,11E)-3,7,11,15-Cembratetraen-17,2-Olide (LS-1) from Lobophyyum sp. on the Apoptosis Induction of SNU-C5 Human Colorectal Cancer Cells

  • Kim, Eun-Ji;Kang, Jung Il;Tung, Nguyen-Huu;Kim, Young-Ho;Hyun, Jin Won;Koh, Young Sang;Chang, Weon-Young;Yoo, Eun Sook;Kang, Hee-Kyoung
    • Biomolecules & Therapeutics
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    • v.24 no.6
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    • pp.623-629
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    • 2016
  • (1S,2S,3E,7E,11E)-3,7,11,15-cembratetraen-17,2-olide (LS-1), a marine cembrenolide diterpene, has anticancer activity against colon cancer cells such as HT-29, SNU-C5/5-FU (fluorouracil-resistant SNU-C5) and SNU-C5. However, the action mechanism of LS-1 on SNU-C5 human colon cancer cells has not been fully elucidated. In this study, we investigated whether the anticancer effect of LS-1could result from apoptosis via the modulation of $Wnt/{\beta}$-catenin and the TGF-${\beta}$ pathways. When treated with the LS-1, we could observe the apoptotic characteristics such as apoptotic bodies and the increase of sub-G1 hypodiploid cell population, increase of Bax level, decrease of Bcl-2 expression, cleavage of procaspase-3 and cleavage of poly (ADP-ribose) polymerase in SNU-C5 cells. Furthermore, the apoptosis induction of SNU-C5 cells upon LS-1 treatment was also accompanied by the down-regulation of $Wnt/{\beta}$-catenin signaling pathway via the decrease of GSK-$3{\beta}$ phosphorylation followed by the decrease of ${\beta}$-catenin level. In addition, the LS-1 induced the activation of TGF-${\beta}$ signaling pathway with the decrease of carcinoembryonic antigen which leads to decrease of c-Myc, an oncoprotein. These data suggest that the LS-1 could induce the apoptosis via the down-regulation of $Wnt/{\beta}$-catenin pathway and the activation of TGF-${\beta}$ pathway in SNU-C5 human colon cancer cells. The results support that the LS-1 might have potential for the treatment of human colon cancer.

Immunomodulating and Antitumor Activities of Panellus serotinus Polysaccharides

  • Kim, Jeong-Hwa;Lee, Jae-Seong;Lee, Kyung-Rim;Shim, Mi-Ja;Lee, Min-Woong;Shin, Pyung-Gyun;Cheong, Jong-Chun;Yoo, Young-Bok;Lee, Tae-Soo
    • Mycobiology
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    • v.40 no.3
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    • pp.181-188
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    • 2012
  • This study was initiated in order to investigate the anticancer and immunomodulating activities of crude polysaccharides extracted in methanol, neutral saline, and hot water (hereinafter referred to as Fr. MeOH, Fr. NaCl, and Fr. HW, respectively) from the fruiting bodies of Panellus serotinus. Content of ${\beta}$-glucan and protein in Fr. MeOH, Fr. NaCl, and Fr. HW extracts of P. serotinus ranged from 22.92~28.52 g/100 g and 3.24~3.68 g/100 g, respectively. In vitro cytotoxicity tests, none of the various fractions of crude polysaccharides were cytotoxic against sarcoma 180, HT-29, NIH3T3, and RAW 264.7 cell lines at the tested concentration. Intraperitoneal injection with crude polysaccharides resulted in a life prolongation effect of 23.53~44.71% in mice previously inoculated with sarcoma 180. Treatment with Fr. HW resulted in an increase in the numbers of spleen cells by 1.3 fold at the concentration of $50{\mu}g/mL$ compared with control. Treatment with Fr. NaCl resulted in improvement of the immuno-potentiating activity of B lymphocytes by increasing the alkaline phosphatase activity by 1.4 fold, compared with control, at the concentration of $200{\mu}g/mL$. Among the three fractions, maximum nitric oxide ($13.48{\mu}M$) was recorded at $500{\mu}g/mL$ in Fr. HW. Production of tumor necrosis factor alpha, interleukin-$1{\beta}$, and interleukin-6 was significantly higher, compared to the positive control, concanavalin A, at the tested concentration. Therefore, treatment with crude polysaccharides extracted from the fruiting body of P. serotinus could result in improvement of antitumor activity.

Modulation of Chemical Stability and Cytotoxic Effects of Epigallocatechin-3-gallate by Different Types of Antioxidants (Epigallocatechin-3-gallate의 화학안정성 및 세포독성에 미치는 각종 항산화제의 영향)

  • Kim, Mi-Ri;Kang, Smee;Hong, Jung-Il
    • Korean Journal of Food Science and Technology
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    • v.43 no.4
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    • pp.483-489
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    • 2011
  • Epigallocatechin-3-gallate (EGCG) is a polyphenolic compound frequently found in green tea, and its physiological actions have been extensively investigated. In the present study, changes in chemical stability and cytotoxic properties of EGCG in the presence of different types of antioxidants were investigated. The antioxidants used modulated the chemical stability of EGCG. Superoxide dismutase (SOD) significantly increased EGCG stability; EGCG was less stable in the presence of catalase. Ascorbic acid, N-acetylcysteine (NAC), and glutathione (GSH) stabilized EGCG concentration dependently. The $H_2O_2$ level generated from EGCG was decreased by catalase, SOD, and NAC but not by GSH. The cytotoxic effects of EGCG also decreased in the presence of NAC, catalase, and SOD. GSH, however, showed a complicated modulatory pattern according to the EGCG and GSH concentrations, and ascorbic acid rather enhanced EGCG toxicity. The results suggest that certain antioxidants could modulate the cytotoxic properties of EGCG in a cell culture system not only by removing reactive oxygen species but by modulating chemical stability and other factors, which should be considered carefully when studying reactive oxygen species-dependent mechanisms of EGCG.

Spray-dried powder preparation of pumpkin sweet potato hydrolysates and its physicochemical properties (호박고구마 효소 분해물의 분무건조 분말 제조 및 물리화학적 품질특성)

  • Lee, Dae-Hoon;Jang, Jong-Hyun;Hong, Joo-Heon
    • Food Science and Preservation
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    • v.24 no.2
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    • pp.246-253
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    • 2017
  • This study was conducted prepare spray-dried powder using pumpkin sweet potato hydrolysates and examine the physicochemical properties of the powder. The insoluble dietary fiber and soluble dietary fiber of the pumpkin sweet potato treated by enzyme were 4.17% and 2.07%, respectively. The spray-dried pumpkin sweet potato hydrolysates was manufactured via spray-drying with different forming agents: i.e., pectin 0.1%, 0.5%, 1%, and 2.0%. The moisture contents and total starches of the spray-dried powders were approximately 1.68-2.46 and 45.32-46.51%, respectively. The color of the L and a value decreased, and that of the b and ${\Delta}E$ value increased. The particle size and outer topology of the spray-dried powders were $37.17-42.32{\mu}m$, and its shape was generally globular. The water absorption index of the spray-dried powder (1.74-1.91) was lower than that of the freeze-dried powder (2.15). The water solubility index of the spray-dried powder, 80.75-87.61%, was higher than that of the freeze-dried powder (70.47%). The adhesion values of spray-dried powder to epithelial HT-29 cells were 2.66-6.18% of the initial cell counts, whereas freeze-dried powder showed lower adhesive ability (1.79%). The in vitro human digestibility in the spray-dried powder was 70.09% which is very effective in digestion.

Biological Activities in the Leaf Extract of Lythrum salicaria L. (털부처꽃 잎 추출물의 생리활성탐색)

  • Kim, Hee-Yeon;Lim, Sang-Hyun;Park, Min-Hee;Park, Yu-Hwa;Ham, Hun-Ju;Lee, Ki-Yun;Park, Dong-Sik;Kim, Kyung-Hee;Kim, Song-Mun
    • Korean Journal of Medicinal Crop Science
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    • v.18 no.6
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    • pp.409-415
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    • 2010
  • In this study, the bioactivities of ethanol (EELS) and water extract (WELS) from the leaf of Lythrum salicaria L. were investigated. In the anti-cancer activity, the growths of both human prostate cancer (DU145) and human colonic carcinoma cell (HT29) were inhibited up 60% by adding 10 mg/$m{\ell}$ of EELS. Anti-inflammatory activity of EELS and WELS have been evaluated on lipopolysaccharide (LPS) induced release of nitric oxide (NO) by the macrophage RAW 264.7 cells. EELS and WELS inhibited inflammatory by 57.3 and 46.9% in 10 mg/$m{\ell}$, respectively. In the anti-oxidative activity, $IC_{50}$ of DPPH radical scavenging activity was respectively 60.71 and $92.90\;{\mu}g/m{\ell}$ by EELS and WELS. In the anti-diabetic activity, $IC_{50}$ of ${\alpha}$-amylase inhibitory activity of EELS and WELS were respectively 5,250 and $5,020\;{\mu}g/m{\ell}$. $IC_{50}$ of ${\alpha}$-glucosidase inhibitory activity was 7.96 and $68.41\;{\mu}g/m{\ell}$ by EELS and WELS. In the anti-obesity, $IC_{50}$ of lipase inhibitory activity was 880 and $9,840\;{\mu}g/m{\ell}$ by EELS and WELS. Finally, EELS and WELS exhibited anti-oxidative, anti-inflammatory, anti-diabetic activity and anti-obesity. It suggests that Lythrum salicaria L. could be potentially used as a resource of bioactive materials for health functional foods.

Synergistic Effect of Yuza(Citrus junos) Extracts and Ascorbic Acid on Antiproliferation of Human Cancer Cells and Antioxidant Activity (비타민 C가 첨가된 유자 추출물의 항산화능과 암세포 증식억제 상승효과)

  • Shon, Mi-Yae;Park, Seok-Kyu
    • Food Science and Preservation
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    • v.13 no.5
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    • pp.649-654
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    • 2006
  • To enhance beneficial effects of citron fruits, anticancer and antioxidant activities of citron fruits extracts were assessed with or without ascorbic acid. Total phenolic acids and flavonoids of fruits peels and flesh extracts were determined. Fruits peels contained more phenolic acids and flavonoids than those detected in flesh extracts. Scavenging activities of 1,1-diphenyl-2-picrylhydrazyl radicals and reducing powers were increased depending on the concentration. The antioxidant activities on oxidation of linoleic acid emulsion incubated at $50^{\circ}C$ were increased but the effect was small to that of butylated hydroxy toluene and ascorbic acid. The anti-tumorigenic effect of these compounds were investigated. They were shown to inhibit the in vitro proliferation of four human tumorigenic cell lines, HT-29, MCF-7, DU-145 and HepG2, in a doso-dependent manner. This study demonstrated that the antioxidant and anticancer activities of citron fruits extracts were derived from their phenols and flavonoids.

The Immuno-Modulatory and Antitumor Effects of Crude Polysaccharides Extracted from Tremella fuciformis (흰목이(Tremella fuciformis)에서 추출한 조다당류의 면역 활성 및 항암 효과)

  • Oh, Yun-Hee;Kim, Sang-Beom;Lee, Gun-Woo;Kim, Hye-Young;Shim, Mi-Ja;Rho, Hyun-Su;Lee, Hyun-Sook;Lee, Min-Woong;Lee, U-Youn;Lee, Tae-Soo
    • The Korean Journal of Mycology
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    • v.34 no.2
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    • pp.105-111
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    • 2006
  • Tremella fuciformis, one of edible and medicinal mushroom belonging to Tremellaceae of Basidiomycota, has been known to have a curative effect on sarcoma 180 of mice and lowering high blood pressure of human beings. Neutral salt soluble [0.9% NaCl (Fr. NaCl)], hot water soluble (Fr. HW) and methanol soluble (Fr. MeOH) substances were extracted from Tremella fuciformis. In vitro cytotoxicity tests, Fr. HW and Fr. NaCl were not cytotoxic against cancer cell lines such as NIH3T3, Sarcoma 180, and HT-29 at the concentration of $2000{\mu}g/ml$, while Fr. MeOH was cytotoxic to NIH3T3 and Sarcoma 180. Intraperitoneal injection with Fr. NaCl showed antitumor effect with life prolongation of 53% in mice inoculated with Sarcoma 180. Fr. NaCl improved the immunopotentiation activity of B lymphocyte by increasing the alkaline phosphatase activity by $3.0{\sim}8.1$ folds, respectively. Intraperitoneal injection with Fr. NaCl increased the numbers of peritoneal exudated cells and circulating leukocytes by 7.4 folds and 1.6 folds, respectively, than in the control group. The antitumor effect of T. fuciformis against Sarcoma 180 of mice was likely due to immunopotentiation activity.

Inhibitory Effects of Solvent Extracts from Seven Brown Algae on Mutagenicity and Growth of Human Cancer Cells (7종 갈조류의 항돌연변이 및 인체 암세포 증식 억제 효과)

  • Choi, Hyung-Ju;Kil, Jeung-Ha;Bak, Soon-Sun;Kong, Chang-Suk;Park, Kun-Young;Seo, Young-Wan;Lim, Sun-Young
    • Journal of Life Science
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    • v.16 no.7 s.80
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    • pp.1080-1086
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    • 2006
  • 본 연구에서는 해조류의 항돌연변이 및 항암 생리활성물질을 검색하여 발암물질 생성 방지 및 생체 방어 물질로서의 이용 가능성을 검토하고자 Ames test를 이용하여 직접돌연변이원인 MNNG와 간접돌연변이원인 $AFB_1$에 대한 항돌연변이 효과 및 암세포 증식 억제 효과를 알아보고자 하였다. $AFB_1$에 대해서 괭생이모자반(S. horneri)이 실험에 사용된 다른 해조류들 중에서 가장 높은 돌연변이 억제 효과를 보였다. 첨가농도 1.25mg/plate일 때, 괭생이모자반의 acetone+methylene chloride 추출물과 methanol 추출물은 각각 96%, 91%로 실험에 사용된 다른 해조류들의 추출물들 중에서 가장 높았으며 양성 대조군인 다시마의 용매 추출물보다도 높은 돌연변이 억제 효과를 보였다. $AFB_1$과 같은 농도인 0.6mg/plate의 농도의 MNNG를 사용하여 S. typhimurium TA100 균주에 대한 해조류의 항돌연변이성 실험을 한 결과, 간접 돌연변이원인 $AFB_1$에 비해 직접 돌연변이원인 MNNG에 대해서는 다소 항돌연변이 효과가 떨어지지만, 여기서도 실험에 사용된 해조류들의 돌연변이 억제 효과를 살펴볼 수 있었으며 acetone+methylene chloride 추출물의 경우가 methanol 추출물보다 다소 높은 활성을 나타내었다. 항돌연변이 실험에서 효과가 뛰어난 괭생이모자반과 짝잎모자반을 중심으로 인체 암세포(위암세포, AGS 및 결장암 세포, HT-29) 증식억제효과를 살펴본 결과, 용매 추출물을 0.5%, 1% 및 2%의 농도별로 암세포에 처리했을 때 acetone+methylene chloride 추출물과 methanol 추출물은 둘 다 가장 낮은 농도인 0.5%에서부터 농도 의존적으로 암세포 증식 억제 효과가 증가하였다. 이상의 7종의 갈조류 추출물들은 Ames test에서 높은 항돌연변이 효과를 나타냈을 뿐만 아니라 괭생이모자반 및 짝잎모자반은 인체 암세포에 대해서도 높은 증식 억제 효과를 나타냄을 살펴 볼 수가 있었다.