• Title/Summary/Keyword: HeLa $S_3$ cells

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Studies on Biological Activities of the Polysaccharides and Oligosaccharides of Orostachys japonicus (와송 다당체 및 올리고당류의 생리 활성)

  • Kim Ki Hoon;Kim Eun Young;Kim Yea Oon;Baek Geum Ok;Kim Han Bok;Lee Dong Seok
    • Korean Journal of Microbiology
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    • v.40 no.4
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    • pp.334-341
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    • 2004
  • Polysaccharides were prepared from Orostachys japonicus by extration with hot steam water (OJPl). The OIPl fraction was further purified by Sephadex G-50 gel filtration chromatography to produce FI (polysaccharides) and FII (oligosaccharides) fraction. The average molecular masses o fFI and FII fraction were determined to be 3050 kDa and 13 kDa, respectively. The antimicrobial activity of OIPl was tested against 8 strains of bacteria and one strain of yeast by the disc diffusion method, fluorescein diacetate (FDA) method and broth dilution method. The OIPl exhibited a very strong growth inhibition to Candida albicans. The OIPl remarkably sup­pressed the growth of Salmonella typhimurium and Staphylococcus aureus. The OIPl showed higher growth inhibition to Escherichia coli and Pseudomonas aeruginosa than propolis, positive control. When the anticancer activity of the OIPl, FI or FII was examined against human cancer cell lines and the Sarcoma 180 cells, these widely suppressed the proliferation of cell lines in the MTT assay and morphology study. Especially, they remarkably inhibited the growth of A549, HeLa and AGS cells. Also treatment of cancer cells with OJPl, FI or FII induced apoptotic cell death characterized by DNA fragmentation. The OJPl, FI or FII exhibiting various biological activities such as antimicrobial activity and anticancer activity is expected to be developed as new biohealth products.

Serum Deprivation Enhances Apoptotic Cell Death by Increasing Mitochondrial Enzyme Activity

  • Moon, Eun-Yi
    • Biomolecules & Therapeutics
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    • v.16 no.1
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    • pp.1-8
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    • 2008
  • Mitochondria are important sensor of apoptosis. $H_2O_2-induced$ cell death rate was enhanced by serum deprivation. In this study, we investigated whether serum deprivation using 0.5 or 3 % FBS induces apoptotic cell death through mitochondrial enzyme activation as compared to 10 % FBS. Apoptotic cell death was observed by chromosome condensation and the increase of sub-G0/G1 population. Serum deprivation reduced cell growth rate, which was confirmed by the decrease of S-phase population in cell cycle. Serum deprivation significantly increased caspase-9 activity and cytochrome c release from mitochondria into cytosol. Serum deprivation-induced mitochondrial changes were also indicated by the increase of ROS production and the activation of mitochondrial enzyme, succinate dehydrogenase. Mitochondrial enzyme activity increased by serum deprivation was reduced by the treatment with rotenone, mitochondrial electron transport inhibitor. In conclusion, serum deprivation induced mitochondrial apoptotic cell death through the elevation of mitochondrial changes such as ROS production, cytochrome c release and caspase-9 activation. It suggests that drug sensitivity could be enhanced by the increase of mitochondrial enzyme activity in serum-deprived condition.

Bioassay-guided Isolation of Deoxypodophyllotoxin, the Cytotoxic Constituent of Juniperus chinensis

  • Ali, A.M.;Intan-Safinar, I.;Mackeen, M.M.;El-Sharkawy, S.H.;Takahata, K.;Kanzaki, H.;Kawazu, K.
    • Natural Product Sciences
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    • v.4 no.3
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    • pp.180-183
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    • 1998
  • The ethanol extract from the leaves of Juniperus chinensis was found to be cytotoxic towards HeLa cells. Bioassay-guided fractionation of the EtOAc soluble faction directed by the microtitration cytotoxic assay revealed that the cytotoxic compound was deoxypodophyllotoxin. All the tumour cell lines tested (KU8112F-chronic mylogeneous leukemia, TK 10-renal carcinoma, UACC 62-melanoma and CEM-SS-T-lymphoblastic leukemia) were found to be susceptible to deoxypodophyllotoxin, however, the minimum effective concentration (MEC) required to reduce the cell population by 100 percent was different between cell lines.

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X-ray Induced Unscheduled DNA Synthesis in Relation to Chromosome Exchange and Mitotic Activity in Established Mammalian Cells (哺乳動物細胞에 있어서 X-線에 의한 回復複製와 染色體交換 및 分裂活動과의 聯關性)

  • Park, Sang-Dai
    • The Korean Journal of Zoology
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    • v.15 no.3
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    • pp.95-100
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    • 1972
  • Dose response and time dependence of unscheduled DNA synthesis induced by X-rays were measured to determine if any correlation exists between unscheduled DNA synthesis, modal chromosome number, chromosome exchange and mitotic activity in four mammalian cell strains. Unscheduled DNA synthesis occurred in all strains studied. The rate was dose-dependent and strain-specific. Only HeLa $S_3$ showed a staturated dose response after 4, 000 R, other cells were linearly proportional to dose increases. Time dependence of unscheduled DNA synthesis was completed within 2 hours after irradiation regardless of cell strains. Unscheduled DNA synthesis was not directly related to modal chromosome number, total exchange rate and mitotic activity. Mitotic activity and chromosome exchange were both dose-dependent, but the rates of them were inversely related.

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Antimutagenic and Antitumor Effects of Codonopsis lanceolata Extracts (더덕 추출물의 항돌연변이 및 항종양 효과)

  • Kim, Soo-Hyun;Choi, Hyun-Jin;Chung, Mi-Ja;Cui, Cheng-Bi;Ham, Seung-Shi
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.10
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    • pp.1295-1301
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    • 2009
  • This study was carried out to investigate the mutagenic, antimutagenic, cytotoxicity and antitumor effect of Codonopsis lanceolata (CL). CL was extracted with 70% ethanol and then further fractionated to hexane, chloroform, ethyl acetate, butanol and water. Antimutagenic, cytotoxicity and antitumor effects of CL extracts were measured by using Ames test, SRB method, and the tumor growth inhibition test. CL extracts did not show any mutagenicity in the Ames test; however, 70% ethanol extracts and its fractions had strong antimutagenic effects against mutation induced by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) and 4-nitroquinoline-1-oxide (4NQO). The ethyl acetate fraction of CL (200 ${\mu}g$/plate) showed approximately 72.1% inhibitory effect on the mutagenesis induced by 4NQO against TA98 strain, whereas 69.6% and 67.0% inhibitions were observed on the mutagenesis induced by MNNG and 4NQO against TA100 strain. In anticancer effects, the cytotoxicity of CL extract and its fractions against cancer cell lines including human cervical adenocarcinoma (HeLa), human hepatocellular carcinoma (HepG2), human breast adenocarcinoma (MCF-7), human lung carcinoma (A549) and transformed primary human embryo kidney (293) were investigated. The treatment of 1 mg/mL CL ethyl acetate fraction had the highest cytotoxicity of 74.5%, 70.7% and 80.3% against HeLa, MCF-7 and A549 cells, respectively. In contrast, the extract and its fractions showed only 2$\sim$31% cytotoxicity for a normal human kidney cell line (293). In vivo anticancer effect of CL extract was tested using Balb/c mice transplanted sarcoma-180 cells. CL ethyl acetate fraction showed the highest inhibition rate of 56.4% at the 50 mg/kg concentration.

Antioxidative and Cytotoxic Effects of Solvent Fractions from Elaeagnus multiflora (뜰보리수 열매의 용매분획별 항산화 및 암세포 증식 억제 효과)

  • Kim, Sung-Ae;Oh, Se-In;Lee, Mee-Sook
    • The Korean Journal of Food And Nutrition
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    • v.20 no.2
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    • pp.134-142
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    • 2007
  • This study was performed to determine the antioxidative and cytotoxic effects of Elaeagnus multiflora by examining its scavenging effects on the 1,1-diphenyl-2-picryl-hydrazyl(DPPH) radical, the inhibition of lipid peroxidation, and its inhibitory effects on cancer cell proliferation in HeLa cells, MCF-7 cells, and SNU-638 cells by MTT assay. Here, dried samples were extracted in ehtanol at room temperature and fractionated into five different solvent types: hexane, dichloromethane, ethylacetate, butanol, and aqueous partition layers. The hexane(62.92${\pm}$2.45%) and dichloromethane(65.25${\pm}$4.74%) fractions of Elaeagnus multiflora's flesh, and the aqueous(94.65${\pm}$0.02%) and ethylacetate(93.83${\pm}$0.02%) fractions of Elaeagnus multiflora's seeds, inhibited DPPH radical production. The DPPH radical scavenging effects of the flesh and seed were different according to solvent fractions. The inhibition of lipid peroxidation by the flesh and seed extracts were 76.11${\pm}$3.66 and 69.57${\pm}$2.27, respectively, for hexane and 67.57${\pm}$2.43 and 62.09${\pm}$0.90, respectively, for the dichloromethane fraction. Among the various partition layers of Elaeagnus multiflora's flesh, hexane and dichloromethane showed the strong cytotoxicities on all the cancer cell lines used in the study. Also all the fractions of Elaeagnus multiflora's seed exhibited significant effects on the inhibition of cancer cell growth(hexane > dichloromethane > ethylacetate > butanol > aqueous partition layers). These results indicate that the haxane and dichloromethane partition layers of Elaeagnus multiflora's flesh and seed extracts have possible antioxidative and anticancer capacities. Although further studies are needed, the present work suggests that Elaeagnus multiflora may be an antioxidative and chemopreventive agent.

Justicidin A Reduces β-Amyloid via Inhibiting Endocytosis of β-Amyloid Precursor Protein

  • Chun, Yoon Sun;Kwon, Oh-Hoon;Oh, Hyun Geun;Cho, Yoon Young;Yang, Hyun Ok;Chung, Sungkwon
    • Biomolecules & Therapeutics
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    • v.27 no.3
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    • pp.276-282
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    • 2019
  • ${\beta}$-amyloid precursor protein (APP) can be cleaved by ${\alpha}$-, and ${\gamma}$-secretase at plasma membrane producing soluble ectodomain fragment ($sAPP{\alpha}$). Alternatively, following endocytosis, APP is cleaved by ${\beta}$-, and ${\gamma}$-secretase at early endosomes generating ${\beta}$-amyloid ($A{\beta}$), the main culprit in Alzheimer's disease (AD). Thus, APP endocytosis is critical for $A{\beta}$ production. Recently, we reported that Monsonia angustifolia, the indigenous vegetables consumed in Tanzania, improved cognitive function and decreased $A{\beta}$ production. In this study, we examined the underlying mechanism of justicidin A, the active compound of M. angustifolia, on $A{\beta}$ production. We found that justicidin A reduced endocytosis of APP, increasing $sAPP{\alpha}$ level, while decreasing $A{\beta}$ level in HeLa cells overexpressing human APP with the Swedish mutation. The effect of justicidin A on $A{\beta}$ production was blocked by endocytosis inhibitors, indicating that the decreased APP endocytosis by justicidin A is the underlying mechanism. Thus, justicidin A, the active compound of M. angustifolia, may be a novel agent for AD treatment.

Isolation of Anticancer Compounds from Peucedanum japonicum Thunb. Roots

  • Jun, Neung Jae;Kim, Seong-Cheol;Song, Eun-Young;Jang, Ki Chang;Lee, Dong Sun;Cho, Somi K.
    • Korean Journal of Plant Resources
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    • v.27 no.3
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    • pp.215-222
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    • 2014
  • This study was conducted to isolate a compound with anticancer properties from the roots of Peucedanum japonicum Thunb. (Umbelliferae), and to evaluate the efficacy of that compound's anticancer activity. The $CHCl_3$ layer was purified via repeated column chromatography and recrystallization. The two compounds isolated from $CHCl_3$ layer were identified via NMR spectroscopic analysis as (10E) 1,10-heptadecadiene-4,6-diyne-3,8,9-triol (Comp. I) and anomalin (Comp. II). (10E) 1,10-heptadecadiene-4,6-diyne-3,8,9-triol was the first report from the roots of P. japonicum. MTT assays were conducted to evaluate the in vitro cytotoxic activities of Compounds I and II against the following human cancer cell lines: HeLa, HepG2, SNU-16, and AGS. Comp. I evidenced the most profound cytotoxic activity against HepG2 cells ($IC_{50}=6.04{\mu}g/mL$), and Comp. II exhibited the most profound cytotoxic activity against SNU-16 cells ($IC_{50}=18.24{\mu}g/mL$) among the human cancer cell lines tested in this study. However, no significant cell death was observed in the CCD-25Lu human normal lung fibroblast cells. Quantitative analysis using UPLC (Ultra Performance Liquid Chromatography) showed that the roots of P. japonicum contained 0.015 (Comp. I) and 1.69 mg/g (Comp. II) of these compounds.

Expression Comparison of the GFP Gene under the Controls of Several Internal Promoters in the Retrovirus Vectors with or without WPRE Sequence (여러 표적세포에서 Retrovirus Vector의 내부 Promoter의 종류와 WPRE의 유무에 따른 GFP 유전자의 발현 효율성 비교)

  • Kim Y. H.;Koo B. C.;Kwon M. S.;Kim T. O.
    • Reproductive and Developmental Biology
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    • v.28 no.3
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    • pp.191-196
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    • 2004
  • In this study, to construct more effective retrovirus system, we compared four internal promoters (RSV: Rous sarcoma virus, UbC: Ubiquitin C, β-actin, CMV: human cytomegalovirus) in the retrovirus vector by measuring GFP (green fluorescent protein) expression. The effect of WPRE (woodchuck hepatitis virus posttrans-criptional regulatory element) sequence on transgene expression was also investigated. Experiments were conducted with cells derived from three different species (human, pig and chicken) and evaluated the activity of each promoter and the effect of WPRE sequence by fluorometry and Western blotting. In all cells tested, RSV and CMV promoters were superior to UbC and β-actin promoters, and RSV promoter was the best one in chicken cells. The boosting effect of WPRE sequence on GFP expression was evident in human and porcine cells but not in chicken cells.

Rapid Identification of Bioactive Compounds Reducing the Production of Amyloid β-Peptide (Aβ) from South African Plants Using an Automated HPLC/SPE/HPLC Coupling System

  • Kwon, Hak-Cheol;Cha, Jin-Wook;Park, Jin-Soo;Chun, Yoon-Sun;Moodley, Nivan;Maharaj, Vinesh J.;Youn, Sung-Hee;Chung, Sung-Kwon;Yang, Hyun-Ok
    • Biomolecules & Therapeutics
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    • v.19 no.1
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    • pp.90-96
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    • 2011
  • Automated HPLC/SPE/HPLC coupling experiments using the Sepbox system allowed the rapid identification of four bioactive principles reducing the production of amyloid $\beta$-peptide ($A{\beta}$) from two South African plants, Euclea crispa subsp. crispa and Crinum macowanii. The structures of biologically active compounds isolated from the methanol extract of Euclea crispa subsp. crispa were assigned as 3-oxo-oleanolic acid (1) and natalenone (2) based on their NMR and MS data, while lycorine (3) and hamayne (4) were isolated from the dichloromethane-methanol (1:1) extract of Crinum macowanii. These compounds were shown to inhibit the production of $A{\beta}$ from HeLa cells stably expressing Swedish mutant form of amyloid precursor protein (APPsw).