• Title/Summary/Keyword: HeLa cell lines

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Fenofibrate decreases radiation sensitivity via peroxisome proliferator-activated receptor ${\alpha}$-mediated superoxide dismutase induction in HeLa cells

  • Liu, Xianguang;Jang, Seong-Soon;An, Zhengzhe;Song, Hye-Jin;Kim, Won-Dong;Yu, Jae-Ran;Park, Woo-Yoon
    • Radiation Oncology Journal
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    • v.30 no.2
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    • pp.88-95
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    • 2012
  • Purpose: The fibrates are ligands for peroxisome proliferator-activated receptor (PPAR) ${\alpha}$ and used clinically as hypolipidemic drugs. The fibrates are known to cause peroxisome proliferation, enhance superoxide dismutase (SOD) expression and catalase activity. The antioxidant actions of the fibrates may modify radiation sensitivity. Here, we investigated the change of the radiation sensitivity in two cervix cancer cell lines in combination with fenofibrate (FF). Materials and Methods: Activity and protein expression of SOD were measured according to the concentration of FF. The mRNA expressions were measured by using real time reverse-transcription polymerase chain reaction. Combined cytotoxic effect of FF and radiation was measured by using clonogenic assay. Results: In HeLa cells total SOD activity was increased with increasing FF doses up to 30 ${\mu}M$. In the other hand, the catalase activity was increased a little. As with activity the protein expression of SOD1 and SOD2 was increased with increasing doses of FF. The mRNAs of SOD1, SOD2, $PPAR{\alpha}$ and $PPAR{\gamma}$ were increased with increasing doses of FF. The reactive oxygen species (ROS) produced by radiation was decreased by preincubation with FF. The surviving fractions (SF) by combining FF and radiation was higher than those of radiation alone. In Me180 cells SOD and catalase activity were not increased with FF. Also, the mRNAs of SOD1, SOD2, and $PPAR{\alpha}$ were not increased with FF. However, the mRNA of $PPAR{\gamma}$ was increased with FF. Conclusion: FF can reduce radiation sensitivity by ROS scavenging via SOD induction in HeLa. SOD induction by FF is related with $PPAR{\alpha}$.

VEGF siRNA Delivery by a Cancer-Specific Cell-Penetrating Peptide

  • Lee, Young Woong;Hwang, Young Eun;Lee, Ju Young;Sohn, Jung-Hoon;Sung, Bong Hyun;Kim, Sun Chang
    • Journal of Microbiology and Biotechnology
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    • v.28 no.3
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    • pp.367-374
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    • 2018
  • RNA interference provides an effective tool for developing antitumor therapies. Cell-penetrating peptides (CPPs) are delivery vectors widely used to efficiently transport small-interfering RNA (siRNA) to intracellular targets. In this study, we investigated the efficacy of the cancer-specific CPP carrier BR2 to specifically transport siRNA to cancer-target cells. Our results showed that BR2 formed a complex with anti-vascular endothelial growth factor siRNA (siVEGF) that exhibited the appropriate size and surface charge for in vivo treatment. Additionally, the BR2-VEGF siRNA complex exhibited significant serum stability and high levels of gene-silencing effects in vitro. Moreover, the transfection efficiency of the complex into a cancer cell line was higher than that observed in non-cancer cell lines, resulting in downregulated intracellular VEGF levels in HeLa cells and comprehensively improved antitumor efficacy in the absence of significant toxicity. These results indicated that BR2 has significant potential for the safe, efficient, and specific delivery of siRNA for diverse applications.

Cytotoxic Effect of Flavonoids from the Roots of Glycyrrhiza uralensis on Human Cancer Cell Lines (감초(Glycyrrhiza uralensis Fisch.)로부터 분리된 flavonoid의 인체 암세포에 대한 세포독성)

  • Park, Ji-Hae;Wu, Qian;Yoo, Ki-Hyun;Yong, Hye-Im;Cho, Sueng-Mock;Chung, In-Sik;Baek, Nam-In
    • Journal of Applied Biological Chemistry
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    • v.54 no.1
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    • pp.67-70
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    • 2011
  • The roots of Glycyrrhiza uralensis Fisch. were extracted with 30% aqueous ethanol (EtOH), and the concentrated extract was partitioned with n-hexane, chloroform ($CHCl_3$), ethyl acetate (EtOAc), n-butanol (n-BuOH), and $H_2O$, successively. From the $CHCl_3$ fraction, four flavonoids were isolated through the repeated silica gel ($SiO_2$), octadecyl silica gel (ODS), and Sephadex LH-20 column chromatographies (c.c.). According to the results of spectroscopic data including nuclear magnetic resonance spectrometry (NMR), electron ionization mass spectrometry (EI/MS), and infrared spectroscopy (IR), the chemical structures of the compounds were determined as glabrol (1), abyssinone II (2), glabridin (3), and isoliquiritigenin (4). The flavonoids were evaluated for cytotoxic effect against human cancer cell lines, HCT-116, HepG2, HeLa, SK-OV-3, SK-BR-3, MCF-7, and SK-MEL-5. Especially, glabrol (1) and glabridin (2) showed $IC_{50}$ values of lower than $25{\mu}M$.

Cytotoxicity of Methanol Extracts from Cudrania tricuspidata Bureau (꾸지뽕나무 메탄올 추출물의 세포독성)

  • Choi, So-Ra;You, Dong-Hyun;Jang, Ik;Ahn, Min-Sil;Song, Eun-Ju;Seo, Sang-Young;Choi, Min-Kyung;Kim, Young-Sun;Kim, Myung-Kon;Choi, Dong-Geun
    • Korean Journal of Medicinal Crop Science
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    • v.20 no.3
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    • pp.153-158
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    • 2012
  • In order to develop as a natural source of anticancer materials of Cudrania tricuspidata, the cytotoxicity of methanol extracts by harvesting parts and times against 8 cell lines including 293 (normal kidney cells) and A-431 (epidermoid carcinoma cells) were investigated using MTT assay. All harvesting parts had hardly cytotoxicity against 293. And methanol extracts of stem bark and root bark showed very high cytotoxicities against 7 cancer cell lines. The cytotoxicity was the highest against HeLa (cervix adenocarcinoma cells) and followed by MCF-7 (breast adenocarcinoma cells), AGS (stomach adenocarcinoma cells), HT-29 (colon adenocarcinoma cells), HepG2 (hepatoblastoma cells), A549 (lung carcinoma cells) and A-431. By the way, leaf extract had a cytotoxicity against only AGS and ripe fruit extract had no cytotoxicity. Among harvesting times, the cytotoxicity of root bark were high from April to September but that of stem bark showed a little difference. These results showed that anticancer activities of Cudrania tricuspidata extracts were eventful changes by harvesting parts and times.

Antioxidant, Antimicrobial, and Cytotoxic Activities of Ovotransferrin from Egg White

  • Moon, Sun-Hee;Lee, Jae-Hoon;Lee, Yong-Ju;Paik, Ji-Yeon;Ahn, Dong-Uk;Paik, Hyun-Dong
    • Food Science of Animal Resources
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    • v.32 no.5
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    • pp.612-617
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    • 2012
  • The antioxidant, antimicrobial, and cytotoxic activities of ovotransferrin were investigated in vitro. The antioxidant capacity of ovotransferrin was evaluated using the 2,2-Diphenyl-1-picryl hydrazyl (DPPH) radical scavenging method, antimicrobial effects using the agar well diffusion method, and cytotoxicity using the 3-(4,5-dimethylthizol-2-yl)-2,5-diphenylatetetrazolium bromide (MTT) assay. The DPPH radical-scavenging capacity of ovotransferrin at 1 mg/mL level reached approximately 60% after 48 h of reaction. The antimicrobial effects of ovotransferrin against common food-borne pathogens, Staphylococcus aureus KCCM 32395, Bacillus cereus KCCM 40935, Listeria monocytogenes ATCC 15313, Escherichia coli O157:H7 ATCC 43895, and Helicobacter pylori HpKCTC 26695 were dose dependant. Gram-positive bacteria was more sensitive to ovotransferrin than gram-negative bacteria. Ovotransferrin showed stronger antimicrobial effect against L. monocytogenes than other gram-positive bacteria tested. The cytotoxicity of ovotransferrin was evaluated in human cancer cell lines, various tissue origins, including the larynx (Hep-2), stomach (AGS), lung (SK-MES-1), liver (HepG2), breast (MCF-7), cervix (HeLa), and colon (HT-29). Ovotransferrin displayed relatively high cytotoxicity (${\leq}60%$ inhibition effects) at 40 mg/mL. At lower concentrations (${\leq}10mg/mL$), however, ovotransferrin cytotoxic effects were not significant in all cancer cell lines tested. These results indicated that ovotransferrin has potential to be used as an antioxidant or antimicrobial agent in foods or a pharmaceutical agent against cancers.

Sesquiterpenoids from the Stem Bark of Aglaia grandis

  • Harneti, Desi;Permatasari, Atika Ayu;Anisshabira, Amallya;Naini, Al Arofatus;Nurlelasari, Nurlelasari;Mayanti, Tri;Maharani, Rani;Safari, Agus;Hidayat, Ace Tatang;Farabi, Kindi;Supratman, Unang;Azmi, Mohamad Nurul;Shiono, Yoshihito
    • Natural Product Sciences
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    • v.28 no.1
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    • pp.6-12
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    • 2022
  • Five sesquiterpenoids, 7-epi-eudesm-4(15)-ene,1β,6α-diol (1), 7-epi-eudesm-4(15)-ene,1β,6α-diol (2), saniculamoid D (3), aphanamol I (4), and 4β,10α-dihydroxyaromadendrane (5), were isolated from the stem bark of Aglaia grandis. The compounds' (1-5) chemical structures were identified by spectroscopic data including, IR, NMR (1H, 13C, DEPT 135°, HMQC, HMBC, 1H-1H COSY), and HRTOFMS, as well as by comparing with the previously reported spectral data. Therefore, this study described the structural elucidation of compounds 1-5 and evaluated their cytotoxic effects against Hela cervical and B16F10 melanoma cells for the first time, but no significant result was discovered.

Development of a Reporter System Monitoring Regulated Intramembrane Proteolysis of the Transmembrane bZIP Transcription Factor ATF6α

  • Kim, Jin-Ik;Kaufman, Randal J.;Back, Sung Hoon;Moon, Ja-Young
    • Molecules and Cells
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    • v.42 no.11
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    • pp.783-793
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    • 2019
  • When endoplasmic reticulum (ER) functions are perturbed, the ER induces several signaling pathways called unfolded protein response to reestablish ER homeostasis through three ER transmembrane proteins: inositol-requiring enzyme 1 (IRE1), PKR-like ER kinase (PERK), and activating transcription factor 6 (ATF6). Although it is important to measure the activity of ATF6 that can indicate the status of the ER, no specific cell-based reporter assay is currently available. Here, we report a new cell-based method for monitoring ER stress based on the cleavage of $ATF6{\alpha}$ by sequential actions of proteases at the Golgi apparatus during ER stress. A new expressing vector was constructed by using fusion gene of GAL4 DNA binding domain (GAL4DBD) and activation domain derived from herpes simplex virus VP16 protein (VP16AD) followed by a human $ATF6{\alpha}$ N-terminal deletion variant. During ER stress, the GAL4DBD-VP16AD(GV)-$hATF6{\alpha}$ deletion variant was cleaved to liberate active transcription activator encompassing GV-$hATF6{\alpha}$ fragment which could translocate into the nucleus. The translocated GV-$hATF6{\alpha}$ fragment strongly induced the expression of firefly luciferase in HeLa Luciferase Reporter cell line containing a stably integrated 5X GAL4 site-luciferase gene. The established double stable reporter cell line HLR-GV-$hATF6{\alpha}$(333) represents an innovative tool to investigate regulated intramembrane proteolysis of $ATF6{\alpha}$. It can substitute active pATF6(N) binding motif-based reporter cell lines.

A Study on Biological Activities of Opuntia humifusa Cladode Extracts (손바닥 선인장 (Opuntia humifusa) 줄기 추출물의 생리활성)

  • Yoon, Min-Sun;Yoo, Jae-Soo;Lee, Keun-Kwang;Kim, Myung-Kon
    • Journal of Applied Biological Chemistry
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    • v.55 no.2
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    • pp.117-121
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    • 2012
  • Biological activities of the hot water and ethanol extracts from Opuntia humifusa cladodes were investigated. 1,1-diphenyl-2-picryl hadrazyl (DPPH) electron donating ability of hot water and ethanol extracts was 79.07 and 82.54%, respectively. Hot water extract generally showed better cytotoxic activity than ethanol extract against each cell line. HeLa and AGS cell lines treated with hot water extract had more than 50% cytotoxic activities. Based on the antimicrobial activities against four microbial strains, both extracts inhibited growth of Staphylococcus aureus KCCM 11593, whereas affected cell growth of three other microorganisms, Escherichia coli (KCCM 11234), Pseudomonas aeruginosa (ATCC 27853), and Salmonella typhimurium (ATCC 11862), in proportion to the concentration of extracts. The inflammatory activities against hot water extract (34.31%) showed higher than that of ethanol extract (25.59%). The effect of extracts on 3T3-L1 preadipocytes differentiation showed that differentiation of treated group with 80 and 100 ${\mu}g/mL$ of hot and ethanol extracts were increased more than treated group with isobutyl methyl xanthine (IBMX) + dexamethasone. These results indicate that the O. humifusa cladodes extracts can be used as a functional material due to their effective biological activities.

Antimutagenic and Cytotoxicity Effects of Agaricus blazei Murill Extracts (아가리쿠스버섯(Agaricus blazei Murill) 추출물의 항돌연변이원성 및 세포독성 효과)

  • Ji, Jeong-Hwan;Kim, Mi-Nam;Choi, Kun-Pyo;Chung, Cha-Kwon;Ham, Seung-Shi
    • Korean Journal of Food Science and Technology
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    • v.32 no.6
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    • pp.1371-1378
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    • 2000
  • This study was performed to determine the antimutagenic and cytotoxic effect of Agaricus blazei Murill methanol extract on Salmonella typhimurium TA98, TA100 and human cancer cell lines using Ames test and cytotoxicity assay, respectively. In Ames test, methanol extract from A. blazei Murill did not exhibit any mutagenicity and most of the samples showed high antimutagenic effects against mutation induced by N-methyl-N'-nitro-N-nitrosoguanidine(MNNG), 4-nitroquinoline-1-oxide(4NQO), 3-amino-1,4-dimethyl-5H-pyrido [4,3-b] indol(Trp-P-1) and $benzo({\alpha})pyrene(B({\alpha})P)$. The methanol extracts of A. blazei Murill$(200\;{\mu}g/plate)$ showed approximately 92.4%, 81.9% and 83.4% inhibitory effect on the mutagenesis induced by 4NQO, Trp-P-1 and $B({\alpha})P$ against TA98 strain, whereas 87.3%, 94.7%. 92.3% and 89.9% inhibitions were observed on the mutagenesis induced by MNNG, 4NQO, Trp-P-1 and $B({\alpha})P$ against TA100 strain. The solvent fractions of methanol extracts from A. blazei Murill except water fraction showed high antimutagenic effects of $70{\sim}90%$ against mutation induced by MNNG, 4NQO. Trp-P-1 and $B({\alpha})P$. In anticancer effects of A. blazei Murill extract and fraction against cancer cell lines including human breast adenocarcinoma(MCF7), human lung carcinoma(A549), human fibrosarcoma(HT1080), human hepatocellular carcinoma(Hep3B), human epitheloid carcinoma(HeLa), human gastric carcinoma(KATO III) and human chronic myelogenous leukemia(K562) were investigated. The treatment of 1 mg/mL A. blazei Murill extracts had the highest cytotoxicity with 91.9% against HeLa, followed by KATO III(88.7%), A549(86.5%) and Hpe3B(84.3%). Whereas 1 mg/mL treatment of A. blazei Murill extracts had only $10{\sim}40%$ cytotoxicity on human normal liver cell (WRL68).

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Quality Characteristics and Biological Activities of Traditionally Fermented Ginseng Wine

  • Yoon, Yoon-Ji;Kim, Na-Young;Rhee, Young-Kyoung;Han, Myung-Joo
    • Food Science and Biotechnology
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    • v.16 no.2
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    • pp.198-204
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    • 2007
  • The objectives of this study were to determine the quality characteristics, antioxidant activity, and cytotoxicity of fermented ginseng wine at each fermentation step. In the first mash with and without ginseng, viable cell counts (total cell, lactic acid bacteria, and yeast) were maximum between 2 to 4 days of fermentation. At the beginning of fermentation, Brix and ethanol contents, and titratable acidity increased, while pH decreased rapidly. At 3 days of fermentation of the second mash with ginseng, the viable cell counts were similar to those without ginseng and then continually decreased. At the end of fermentation, the pH of the second mash with ginseng was 4.00, lower than the pH of the second mash without ginseng, which was 4.35. Alcohol contents of second mashes with and without ginseng were 12.2 and 11.8%, respectively. In the aging period of ginseng and rice wines, the pH, titratable acidity, Brix, and ethanol contents did not change markedly. The results of sensory evaluation showed that fermented ginseng wine had good flavor and high acceptability. In the 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity assay, fermented ginseng wine ($IC_{50}$: 0.394 mg/mL) showed higher antioxidant activity than fermented rice wine ($IC_{50}$: 0.884 mg/mL). The butanol fraction of fermented ginseng wine exhibited weak cytotoxic activity against P388 and HeLa cell lines.