• Title/Summary/Keyword: Hela cells

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Proteomics Analysis of Immunoprecipitated Proteins Associated with the Oncogenic Kinase Cot

  • Wu, Binhui;Wilmouth, Rupert C.
    • Molecules and Cells
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    • v.25 no.1
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    • pp.43-49
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    • 2008
  • Cancer Osaka thyroid, also known as Tpl-2 (Cot) is a member of the MAP3K kinase family and plays a key role in the regulation of the immune response to pro-inflammatory stimuli such as lipopolysaccharide (LPS) and tumour necrosis $factor-{\alpha}$ ($TNF-{\alpha}$). A series of Cot constructs with an N-terminal 6xHis tag were transiently expressed in HEK293 cells: $Cot_{130-399}$ (kinase domain), $Cot_{1-388}$ (N-terminal and kinase do-mains), $Cot_{1-413}$, $Cot_{1-438}$ (containing a putative PEST sequence), $Cot_{1-457}$ (containing both PEST and degron sequences) and $Cot_{1-467}$ (full-length protein). These Cot proteins were pulled down using an anti-6xHis antibody and separated by 2D electrophoresis. The gels were silver-stained and 21 proteins were detected that did not appear, or had substantially reduced intensity, in the control sample. Three of these were identified by MS and MS/MS analysis as Hsp90, Hsp70 and Grp78. Hsp90 appeared to bind to the kinase domain of Cot and this interaction was further investigated using co-immuno-precipitation with both overexpressed Cot in HEK293 cells and endogenous Cot in Hela cells.

Combined Effect of Radiation and 25-Hydroxycholesterol on Human Cervix and Lung Cancer Cells in vitro

  • Chae Sungwook;Kang Kyoung Ah;Lee Kyoung Hwa;Zhang Rui;Jung Myung Sun;Hyun Jin Won
    • Environmental Mutagens and Carcinogens
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    • v.25 no.2
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    • pp.76-81
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    • 2005
  • 25-Hydroxycholesterol (cholest-5-ene-3, 25-diol, 25-OHC) showed the cytotoxicity on HeLa human cervix and NCI-H460 human lung cancer cells, $0.5{\mu}M\;of\;50\%$ inhibitory concentration. We studied 25-OHC as the possibility of radiation sensitizer. The combination effect of 25-OHC and y-irradiation measured using flow cytometer with propidium iodide stained cells. The combined treatment of 25-OHC and $\gamma-irradiation$ did not show significant enhancing effects on Hela and NCI-H460 cells.

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Effects of Punica granatum L. Fractions on Quinone Reductase Induction and Growth Inhibition on Serveral Cancer Cells (석류 추출성분이 암세포 증식 억제와 Quinone Reductase 유도활성에 미치는 효과)

  • 심선미;최상원;배송자
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.30 no.1
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    • pp.80-85
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    • 2001
  • Various lines of evidence suggest that dietary components protect the initiation step of carcinogenesis. In this study, the ethylacetate (PGMEA), ethylether (PGMEE), butanol (PGMB) and aqueous (PGMA) soluble fractions of Punica granatum L. (PG) were screened for their growth inhibition using the MTT assay on HepG2, HeLa, C6, MCF-7 and HT-29 cells and for their activity to induce quinone reductase (QR) in HepG2 cells. Among various fractions of Punica granatum L., the PGMEE showed the strongest growth inhibition at 500 $\mu\textrm{g}$/mL which resulted 92.5% on Hela cell lines and 97.8% on C6 cell lines. The PGMEA and PGMB also showed significant growth inhibition. The assay of QR induction on HepG2 cells, grown in the presence of PGMEE at the concentration of 50$\mu\textrm{g}$/mL, was 1.4 times more effective compared with the control value of1.0. These results suggested that useful cancer chemoprevention materials could be isolated from PGMEE fraction of Punica granatum L.

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Effect of Extracts from Oriental Plum (Formosa, Oishiwase, Soldam) on LPS-stimulated Raw 264.7 Cells (자두(후무사, 대석, 피자두) 추출물이 LPS로 염증을 유발한 Raw 264.7 세포와 암 세포에 미치는 영향)

  • Kim, Se-Na;Kim, So-Young;Kim, Jung-Bong;Park, Hong-Ju;Cho, Young-Sook
    • Journal of the East Asian Society of Dietary Life
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    • v.23 no.2
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    • pp.197-202
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    • 2013
  • The objective of this study is to evaluate the anti-cancer and anti-inflammatory activities of plum (Formosa, Oishiwase, Soldam) for the future development of functional food products. To determine the anti-inflammatory effect of different types of plums, the inhibitory effect of plum extracts on nitric oxide (NO) production were measured in lipopolysaccharide (LPS)-stimulated Raw 264.7 mouse macrophage cells and human cancer cell lines (A549, Ags, Hela, Hep3B). Among the three different plum cultivars, Oishiwase at a concentration of 1 mg/mL showed the highest inhibitory effects on NO production (%) in Raw 264.7 macrophage cells. Moreover, Oishiwase exhibited a higher anti-cancer activity against A549 (renal carcinoma, 50%), Ags (gastric carcinoma, 35%), HeLa (cervical carcinoma, 50%), and Hep3B (hepatocellular carcinoma, 31%) at a concentration on 1 mg/mL, respectively, compared to Formosa and Soldam. Our findings suggest Oishiwase plum extracts may serve as potential dietary sources of natural health promoting substances.

Studies on the Population of Toxigenic Fungi in Foodstuffs (Ⅶ) Toxicological approches to the Penicillium sp. metabolites isolated from foodstuffs (한국 식품중의 유독성 진균에 관한 연구(제7보) Penicillium 속의 유독성에 대하여)

  • Ko, Chun-Myung;Kim, Sung-Kwang;Cho, Se-Hoon;Kim, Se-Jong;Choi, Tae-Ju;Ryu, Jun
    • Microbiology and Biotechnology Letters
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    • v.2 no.1
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    • pp.19-27
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    • 1974
  • Thirty one culture filtrates of Penicillium app. isolated from foodstuffs were submitted for toxicity by use of HeLa cell and ICR-mice. Nine strains among the 31 of Penicillia were cytotoxic to the HeLa cell cultures. Seven strains among the 31 of Penicillia were toxic to the ICR-mice and the pathological findings were the liver injury featured by parenchymal cell necrosis and degeneration. As a mass screening, cytotoxicity test using HeLa cells is feasible method to detect the mycotoxin-producing fungi.

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Experimental Studies on Antimetastatic and Immunomodulating Effects of Ulmus davidiana (유근피(楡根皮)의 선천 면역 활성화에 의한 암 전이 억제 효과)

  • Kim, Heung-Soo;Cho, Jung-Honn;Lee, Jin-Moo;Lee, Chang-Hoon;Jang, Jun-Bock;Lee, Kyung-Sub
    • The Journal of Korean Obstetrics and Gynecology
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    • v.23 no.1
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    • pp.1-11
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    • 2010
  • Purpose: This study was designed to investigate the antimetastatic and immunomodulating effects of extracts of Ulmus davidiana extracts(U. D. Ex.). Methods: Antimetastatic experiments were conducted in vitro and in vivo by using colon 26-M3.1 carcinoma, L5178Y-R lymphoma cell and Hela cell. To observe the immunomodulating effects of U. D. Ex., we measured IL-6, IL-10, IL-12 and TNF-$\alpha$ from peritoneal macrophages. And we evaluated the activation of NK cell by using anti-asialo-GM1 serum. Results: We found that the administration of U. D. Ex. significantly inhibited tumor metastasis in vivo. In vitro cytotoxicity analysis, cell growth are closer to 100% in case of Colon 26-M3.1 carcinoma, L5178Y-R lymphoma cell and Hela cell at low concentration. In case of macrophage, cell proliferation is closer to 100% less than $250{\mu}g/ml$ of U. D. Ex.. The level of cytokine such as IL-6, IL-10, IL-12 which stimulates U. D. Ex. was increased in dose-dependent manner compared to the control group. In case of TNF-$\alpha$, the level was increased at concentration of $1,000{\mu}g/ml$. The depletion of NK cells by anti-asialo GM1 serum partly abolished the inhibitory effect of U. D. Ex. on tumor metastasis. Conclusion: Ulmus davidiana appears to have considerable activity on the anti-metastasis by activation the immune system.

Effect of Prunellae Spica Extracts on Anti-tumor Metastasis by Immune Activity (면역활성(免疫活性)에 의한 하고초(夏枯草)의 암전이(癌轉移) 억제(抑制) 효과(效果))

  • Heo, Ja-Kyung;Lee, Jin-Moo;Lee, Chang-Hoon;Cho, Jung-Hoon;Jang, Jun-Bok;Lee, Kyung-Sub
    • The Journal of Korean Obstetrics and Gynecology
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    • v.22 no.1
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    • pp.41-52
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    • 2009
  • Purpose: This study was designed to investigate the anti-tumor metastasis by immunomodulating effects of extracts of Prunellae Spica. Methods: Antimetastatic experiment was conducted in vivo by using colon 26-M3.1 carcinoma. And we observed cytotoxicity of Prunellae Spica on colon 26-M3.1 carcinoma, L5178Y-R lymphoma cell, hela cell and macrophage. To observe the immnomodulating effects of Prunellae Spica, we estimated IL-6, IL-10, IL-12, TNF-${\alpha}$ from peritoneal macrophages. And we evaluated the activation of NK cell by using anti-asialo-GM1 serum. Results: We found that the administration of Prunellae Spica extracts significantly inhibited tumor metastasis in vivo. In an in vitro cytotoxicity analysis, cell growth are closer to 100% in case of colon 26-M3.1 carcinoma, L5178Y-R lymphoma cell, hela cell at low concentration. In case of macrophage, cell proliferation is closer to 100% less than $62.5{\mu}g/m{\ell}$ of Prunellae Spica extracts. The level of cytokine such as IL-6, IL-10, IL-12 which stimulates Prunellae Spica extracts was increased in dose-dependent manner compared to the control group. TNF-${\alpha}$ is hardly secreted less than $250{\mu}g/m{\ell}$ The depletion of NK cells by anti-asialo GM1 serum partly abolished the inhibitory effect of Prunellae Spica on tumor metastasis. Conclusion: Prunellae Spica appears to have considerable activity on the anti-metastasis by activation the immune system such as macrophage and NK cell.

Pharmacology of enantiomers of higenamine and related tetrahydroisoquinolines

  • Park, Min-Kyu;Huh, Ja-Myung;Lee, Young-Soo;Kang, Young-Jin;Seo, Han-Geuk;Lee, Jae-Heun;Park, Hye-Sook-Yun-;Lee, Duck-Hyung;Chang, Ki-Churl
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 2004.04a
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    • pp.3-10
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    • 2004
  • Oxidative stress is a constant threat to all living organisms and an immense repertoire of cellular defense systems is being employed by most pro- and eukaryotic systems to eliminate or to attenuate oxidative stress. Ischemia and reperfusion is characterized by both a significant oxidative stress and characteristic changes in the antioxidant defense. Heme oxigenase-l (HO-l) is up-regulated by various stimuli including oxidative stress so that it is thought to participate in general cellular defense mechanisms against ischemic injury in mammalian cells. Higenamine, an active ingredient of Aconite tuber, has been shown to have antioxidant activity along with inhibitory action of inducible nitric oxide synthase (iNOS) expression in various cells. In the present study, we investigated whether higenamine and related analogs protect cells from oxidative cellular injuries by modulating antioxidant enzymes, such as HO-l, MnSOD etc. R-form of YS-51 was the most potent inducer of HO-l in bovine endothelial cells, which inhibited apoptotic cell death by H$_2$O$_2$. HO-1 induction by YS 51 was mediated by PI3 kinase activation in which PKA- as well as PKG pathway is considered as important regulators. YS-51 also induced Mn-SOD mRNA expression by activating c-jun N-terminal kinase in endothelial cells and Hela cells. In ROS 17/2.1 cells, higenamine and enetiomers of related compounds inhibited iNOS expression by cytokine mixtures. Taken together, higenamine and related compounds can be developed as possible protective agents from oxidative cell injury or death.

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Antioxidative and Cytotoxic Effects of Ethanol Extracts from Elaeagnus multiflora (뜰보리수 에탄올 추출물의 산화적 스트레스 억제 효과와 암세포 증식 억제 효과)

  • Oh, Se-In;Lee, Mee-Sook
    • The Korean Journal of Food And Nutrition
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    • v.21 no.4
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    • pp.403-409
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    • 2008
  • Elaeagnus multiflora, generally referred to as the cherry silverberry, is a plant. Elaeagnus multiflora fruit, leaves, and roots have been traditionally utilized in China as a treatment for cough, diarrhea, itch, and foul sores, and even cancer. More importantly, it is being investigated as a food that is capable of reducing the incidence of cancer, and also as a means of halting or reversing the growth of cancers. Considering the dearth of information regarding the medicinal properties of Elaeagnus multiflora, we assessed the antioxidative and cytotoxic effects of Elaeagnus multiflora by examining its scavenging effects on the 1,1-diphenyl-2-picryl-hydrazyl(DPPH) radical, its inhibitory effects on lipid peroxidation, and its inhibitory effects on cancer cell proliferation in HeLa cells, MCF-7 cells, and SNU-638 cells via MTT assay. Ethanol extracts of Elaeagnus multiflora flesh and seed inhibited DPPH radical production($36.91{\pm}1.00$, $94.17{\pm}0.37$) and lipid peroxidation ($28.18{\pm}5.02$, $40.30{\pm}1.45$, respectively). The Elaeagnus multiflora seed is believed to exert a possible antioxidative effects against the DPPH radical. The ethanol extracts of Elaeagnus multiflora flesh and seed exerted the cytotoxic effects on Hela cells ($6.93{\pm}1.92$, $84.97{\pm}0.47$), MCF-7 cells($5.45{\pm}0.41$, $84.97{\pm}0.47$), and SNU-638 cells($19.39{\pm}0.43$, $76.84{\pm}0.63$) used in this study. This result suggests that Elaeagnus multiflora seeds in contrast to its flesh, is believed to exert a possible anticancer effect. Elaeagnus multiflora seeds are considered to be a the candidate for preventative and dietetic treatment as an anticancer functional food.

Study on the Alkaloids from Thalictrum fauriei

  • Chen, Chung-Hsiung
    • Korean Journal of Pharmacognosy
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    • v.17 no.1
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    • pp.49-54
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    • 1986
  • To explore the biological activities of Thalictrum alkaloids, a study was initiated to investigate the alkaloidal constituents from T. fauriei, a species indigenous to Taiwan. Extensive fractionation of the ethanolic extract provided three known aporphines, (+)-oconovine (Ia), (+)-isocorydine (Ib), and (+)-corydine (Ic) from nonphenolic tertiary base fraction. From quaternary base fraction two new protoberberinium salts, thalifaurine (IIa), dehydrodiscretine (IIb), in addition to magnoflorine (III), were isolated. The structure of both new compounds were confirmed by total syntheses. Fractionation of phenolic bases yielded a dihydromorphinandienone, (-)-ocobotrine (IV), as well as a novel aporphine-pavine dimer, tentatively named EP-10. The structure of EP-10 was established by means of 2D NMR studies. Preliminary study indicated a weak activity of lyzing Hela cells, in vitro, for EP-10.

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