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Synergistic Effect of Ethaselen and Selenite Treatment against A549 Human Non-small Cell Lung Cancer Cells

  • Xu, Wei;Ma, Wei-Wei;Zeng, Hui-Hui
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.17
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    • pp.7129-7135
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    • 2014
  • Background: In this study, we aimed to evaluate the growth inhibitory effect of the combination of ethaselen (BBSKE) and low fixed dose of selenite against A549 human non-small cell lung cancer cells in vitro. Materials and Methods: Growth inhibitory effects against A549 cells were determined by SRB assay. Combination index (CI) values were calculated based on Chou-Talalay median-effect analyses. Dose reduction index (DRI) values were applied to calculate dose reduction of selenite. Contents of free thiols and GSH were determined by DTNB assay and intracellular ROS levels by DCFH-DA fluorescence labeling. Results: Compared with BBSKE or selenite single treatment, the combined application of ethaselen and a low fixed dose of selenite shortened the onset time of sodium selenite, reduced $IC_{50}$ values, and increased the maximum inhibition rates, suggesting a possible molecular mechanism of the synergism. Obvious synergistic effects were observed after different times of combination treatment, especially after 24 h. Compared with selenite single treatment, dosage of selenite could be remarkably reduced in combination therapy to gain the same inhibitory effect on cell proliferation. Compared with BBSKE single treatment, the content of free thiols and GSH were significantly reduced and ROS levels greatly elevated in the combination group. For the combination treatment, cell viability increased as greater concentrations of GSH were added. Conclusions: All these results indicate that the combination treatment of BBSKE and selenite showed synergism to inhibit A549 cell proliferation in vitro, and also reduced the selenite dosage to mitigate its toxicity which is very meaningful for combination chemotherapy of lung cancer. The synergism was probably caused by the accelerated exhaustion of intracellular reductive substances, such as free thiols and GSH, which ultimately leads to enhanced oxidative stress and apoptosis.

Cytokinetic Study of MCF-7 Cells Treated with Commercial and Recombinant Bromelain

  • Fouz, Nour;Amid, Azura;Hashim, Yumi Zuhanis Has-Yun
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.11
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    • pp.6709-6714
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    • 2013
  • Background: Breast cancer is a leading cause of death in women. The available chemotherapy drugs have been associated with many side effects. Bromelain has novel medicinal qualities including anti-inflammatory, anti-thrombotic, fibrinolytic and anti-cancer functions. Commercially available bromelain is obtained through tedious methods; therefore, recombinant bromelain may provide a cheaper and simpler choice with similar quality. Materials and Methods: This study aimed to assess the effects of commercial and recombinant bromelain on the cytokinetic behavior of MCF-7 breast cancer cells and their potential as therapeutic alternatives in cancer treatment. Cytotoxic activities of commercial and recombinant bromelain were determined using (sulforhodamine) SRB assay. Next, cell viability assays were conducted to determine effects of commercial and recombinant bromelain on MCF-7 cell cytokinetic behavior. Finally, the established growth kinetic data were used to modify a model that predicts the effects of commercial and recombinant bromelain on MCF-7 cells. Results: Commercial and recombinant bromelain exerted strong effects towards decreasing the cell viability of MCF-7 cells with $IC_{50}$ values of 5.13 ${\mu}g/mL$ and 6.25 ${\mu}g/mL$, respectively, compared to taxol with an $IC_{50}$ value of 0.063 ${\mu}g/mL$. The present results indicate that commercial and recombinant bromelain both have anti-proliferative activity, reduced the number of cell generations from 3.92 to 2.81 for commercial bromelain and to 2.86 for recombinant bromelain, while with taxol reduction was to 3.12. Microscopic observation of bromelain-treated MCF-7 cells demonstrated detachment. Inhibition activity was verified with growth rates decreased dynamically from 0.009 $h^{-1}$ to 0.0059 $h^{-1}$ for commercial bromelain and to 0.0063 $h^{-1}$ for recombinant bromelain. Conclusions: Commercial and recombinant bromelain both affect cytokinetics of MCF-7 cells by decreasing cell viability, demonstrating similar strength to taxol.

The Development of Estimation Model for Iron Compound Originated from Anaerobic Microbial Corrosion (혐기성 미생물부식에 의해 생성되는 철화합물 추정식 개발)

  • Jeon, Seok-Jun
    • Journal of Korean Society on Water Environment
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    • v.18 no.4
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    • pp.379-386
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    • 2002
  • In this study, estimation model for iron compound originated from upflow, anaerobic fixed bed reactor, which treats sewage domestic wastewater, was developed. The estimation model was formulated by a mathematical expression which was based on the mass balance. Below the HRT of 60 minute, sulfide concentration combining with iron $FeS_2$ is the highest because the maximum sulfate consumption rate $V_{maxS}$ and half-saturation constant of sulfate $K_{mS}$ exert an important effect on the estimation model as temperature was increased. But increment of $FeS_2$ concentration is weakened above the HRT of 60 minutes and represent the lowest value at the HRT of 108 minutes. It implies that liquid phase distribution ratio of sulfide ${\alpha}r$ becomes lower as temperature was increased. While phosphorus concentration combining with iron $Fe_3(PO_4)_3$ is increased as HRT and temperature are increased, which is affected by phosphorus removal rate constant $k_p$. As the result of estimating the iron concentrations of corrosion by the model, the concentration of iron corrosion is higher than any other at the HRT of 108 minute and $20^{\circ}C$. The predicted values were compared with measured ones at different HRT(13.5, 27, 54, 108 min) and temperature(20, 25, $30^{\circ}C$). The experimental data could be fitted with the simulated curves. Therefore, the mathematical expression could be applicable to design full-scale wastewater treatment plants.

A Study on the Mechanism of Oxidative Stress, Screening of Protective Agents and Signal Transduction of Cell Differentiation in Cultured Osteoblast and Osteoclast Damaged by Reactive Oxygen Species

  • Park Seung-Taeck;Jeon Seung-Ho
    • Biomedical Science Letters
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    • v.11 no.3
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    • pp.319-326
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    • 2005
  • It is well known that oxidative stress of reactive oxygen species (ROS) may be a causative factor in the pathenogenesis of bone disorder on osteoblast or osteoclast. The purpose of this study was to evaluate the cytotoxicity of oxidative stress, protective effect of glutamate receptor antagoinst against ROS-induced osteotoxicity, secretion of tumor necrosis factor $(TNF)-\alpha$ and the expression of c-fos gene in the cultured rat osteoblasts and osteoclasts. Cell viability by MTS assay or !NT assay, activity of glutathione peroxidase (GPx), lipid peroxidation (LPO) activity, protein synthesis by sulforhodamine B (SRB) assay, alkaline phosphatase (ALP) activity, lactate dehydrogenase (LDH) activity, MTS assay for NMDA (N-methyl-D-aspartate) receptor antagonist or AMPA/kainate receptor antagonist, measurement for $TNF-\alpha$, and c-fos gene expression were performed after these cells were treated with or without various cocentrations of xanthine oxidase (XO), hypoxanthine (HX), D-2-amino-5-phosphonovaleric acid (APV), 7-chlorokynurenic acid (CKA), 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) and 6,7-dinitroquinoxaline-2,3-dione (DNQX), respectively. In this study, XO/HX showed decreased cell viability and glutathione peroxidase (GPx) activity, but it showed increased LPO activity, $TNF-\alpha$ secretion and c-fos expression. APV and CKA incresed protein sythesis and ALP activity. While, CNQX or DNQX did not show any protective effect in LDH activity or cell viability. From these results, XO/HX showed cytotoxic effect in cultured rat osteoblast or osteoclast, and also NMDA receptor antagonist such as APV or CKA was effective in blocking XO/HX-induced osteotoxicity in these cultures.

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Effects of Herbar Chelidonii on the Cultured Spinal Sensory Neurons Damaged by XO/HX (백굴채(白屈菜)가 손상된 배양척수감각신경세포에 미치는 영향)

  • Shin, Byung-Cheul;Song, Yung-Sun
    • The Journal of Korea CHUNA Manual Medicine
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    • v.2 no.1
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    • pp.143-157
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    • 2001
  • Objectives and Methods : To evaluate the mechanism of oxidative damage by xanthine oxydase(XO) and hypoxanthine(HX)-induced oxygen radicals, MTT assay and NR assay were carried out after the cultured mouse spinal sensory neurons were preincubated for 4 hours with various concentrations of XO/HX. And the amount of total protein. neurofilament EIA. lipid peroxidation and LDH activity were measured, to evaluate the protective effect of Herbar Chelidonii(HC) water extract on cultured spinal sensory neurons damaged by XO/HX. after the cultured mouse spinal sensory neurons were preincubated with various concentrations of HC water extract for 3 hours prior to exposure of XO/HX. Results : XO/HX decreased significantly the survival rate of the cultured mouse sensory neurons by NR assay and MTT assay In proportion to concentration and exposed time. In proportion to concentration and exposed time on cultured spinal sensory neurons, XO/HX showed the quantitative decrease of neurofilament by EIA. the decrease of total protein amount by SRB assay and the Increase of lipid peroxidation as well as LDH. HC showed the quantitative increase of neurofilament and total protein, but showed the decrease of lipid peroxidation and LDH activity against the neurotoxicity of XO/HX. Conclusions : From the above results, it is concluded that XO/HX have a neurotoxic effect on cultured spinal sensory neurons and that the herbs extract, such as HC, prevent the toxicity of XO/HX effectively in that they decrease lipid peroxidation and LDH activity.

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New HDAC inhibitor, IN2001 induces apoptosis/cell cycle arrest in human breast cancer cells

  • Euno, Joung-Ki;Nan, Min-Kyung;Jung, Cho-Min;Young, An-Jin;Kim, -Dae-Ki;Yhong, Sheen-Yhun
    • Proceedings of the Korea Society of Environmental Toocicology Conference
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    • 2003.10a
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    • pp.180-180
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    • 2003
  • The acetylation of histone is one of the mechanisms involved in the regulation of gene expression and is tightly controlled by two core enzymes, histone acetyltransferase (HAT) and deacetylase (HDAC). There are several reports that imbalance of HAT and HDAC activity is associated with abnormal behavior of the cells in morphology, cell cycle, differentiation, and carcinogenesis. Recently, an increasing number of structurally diverse HDAC inhibitors have been identified that inhibit proliferation and induce differentiation and/or apoptosis of tumor cells in vivo and in vitro. In this study, we have investigated the effects of novel HDAC inhibitors, IN2001 on ER positive and ER negative human breast cancer cell lines. The growth inhibition, cell cycle arrest and apoptosis of cells by HDAC inhibitors were determined using SRB assay, DNA fragmentation, and flow cytometry. We found that IN 2001 as well as Trichostatin A inhibited cell growth dose-dependently in both ER Positive and ER negative human breast cancer cell lines. The growth inhibition with HDAC inhibitors was associated with profound morphological change. The result of cell cycle analysis after 24 h exposure of IN2001 showed G2-M cell cycle arrest in MCF-7 cell and apoptosis in T47B and MDA-MB-231 cell. In summary, IN2001 has antiproliferative effect on human breast cancer cells regardless of the expression of estrogen receptor. These findings heights the possibility of developing HDAC inhibitors as potential anticancer therapeutic agents for the treatment of breast cancer.

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Treatment of Acid Mine Drainage Using Immobilized Beads Carrying Sulfate Reducing Bacteria (황산염환원균 고정화 담체를 이용한 산성광산배수 처리)

  • Kim, Gyoung-Man;Hur, Won;Baek, Hwan-Jo
    • Economic and Environmental Geology
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    • v.41 no.1
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    • pp.57-62
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    • 2008
  • The application of constructed subsurface-flow wetlands for treatment of wastewater from abandoned mines is being increased. Crushed limestone, oak chips, and mushroom composites are often employed in a bulk form, as the substrates in the bed media. Efficiency of the subsurface-flow treatment system drops with time as the hydraulic conductivity of the wetland soil decreases significantly, presumably due to chemical reactions with the wastewater. The purpose of this study is to investigate the applicability of immobilized beads carrying sulfate reducing bacteria for acid mine drainage treatment system. The ingredients of immobilized beads are organic materials such as mushroom composite and oak chips, limestone powder for a pH buffer, mixed with a modified Coleville Synthetic Brine. It was found that immobilized beads are more efficient than the bulk form for pH recovery, sulfate and heavy metal removal.

Comparative Antitumor Activity of Different Solvent Fractions from an Auricularia auricula-judae Ethanol Extract in P388D1 and Sarcoma 180 Cells

  • Reza, Ahsanur;Choi, Myung-Jin;Damte, Dereje;Jo, Woo-Sik;Lee, Seung-Jin;Lee, Joong-Su;Park, Seung-Chun
    • Toxicological Research
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    • v.27 no.2
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    • pp.77-83
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    • 2011
  • The objective of this study was to evaluate and compare the antitumor activity of different solvent fractions (ethanol, dichloromethane, ethyl acetate, butanol and water) of the Auricularia auricula-judae 70% ethanol extract on the P388D1 macrophage and sarcoma 180 cells. A dose-dependent antitumor activity of each solvent fraction (from 0.01 mg/ml to 0.3 mg/ml) was shown against both cell types. These cytotoxic effects of all the tested fractions were confirmed on the MTT and SRB assays, without statistical differences each other. $IC_{50}$ value of dichloromethane fraction was 94.2 ${\mu}g/ml$ against sarcoma 180 cells lower than any other solvent fractions. The potent antitumor effect of the dichloromethane (DCM) fraction was also found against solid tumor in BALB/c mice. The splenomegaly and higher splenic index were found in tumor-bearing mice, with the DCM fraction returning to the negative control values. Thus, the results indicated the dichloromethane fraction may have potential ingredients as antitumor candidates.

Phenolic Compounds from the Twigs of Corylopsis coreana Uyeki and Their Cytotoxic Activity (히어리나무의 페놀성 화합물 및 세포독성활성)

  • Kwon, Oh Kil;Kim, Chung Sub;Suh, Won Se;Park, Kyoung Jin;Cha, Joon Min;Choi, Sang Un;Kwon, Hak Cheol;Lee, Kang Ro
    • Korean Journal of Pharmacognosy
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    • v.47 no.1
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    • pp.1-6
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    • 2016
  • Phytochemical investigation of the twigs of Corylopsis coreana afforded 10 phenolic compounds, bergenin (1), 6'-O-galloylbergenin (2), 3'-O-galloylbergenin (3), (-)-catechin (4), (-)-epicatechin (5), (-)-epicatechin-3-O-galloyl ester (6), 4-methoxy-3,-5-dihydroxybenzoic acid (7), gallic acid (8), 2,4,6-trimethoxyphenol-1-O-${\beta}-\small{D}$-glucopyranoside (9), and 2,4,6-trimethoxyphenol-1-O-${\beta}-\small{D}$-(6-O-galloyl)-glucopyranoside (10). Their structures were characterized by spectroscopic data and identified by comparing these data with those in the literatures. The compounds 3, 9 and 10 were isolated for the first time from this source. All the isolates (1-10) were tested for their cytotoxic activity against A549, SK-OV-3, SK-MEL-2, and HCT15 cell lines in vitro using the SRB bioassay. The compounds 5, 7 and 8 exhibited selective cytotoxic activity against SK-MEL-2 cell line.

Apoptotic Effect of Vitisin A from Vitis Amurensis against MES-SA Uterine Cancer Cells (왕머루 포도에서 분리한 Vitisin A의 자궁암주에 대한 자멸사 효과)

  • Lim, Jeong-Han;Lee, Hyo-Jeong;Lee, Eun-Ok;Lee, Hyo-Jung;Kwon, Hee-Young;Shim, Bum-Sang;Ahn, Kyoo-Seok;Kim, Sung-Hoon
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.22 no.2
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    • pp.290-295
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    • 2008
  • The cytotoxic characteristics of Vitsin A isolated from Vitis amurensis L. were examined in human colorectal, breast, uterine and renal cancer cells. Vitsin A showed good cytotoxicity against various cancer cells with $IC_{50}$ of $1\;{\sim}\;30\;{\mu}M$. Among them, Vitisin A exhibited strongest cytotoxic effect against MES-SA cells with $IC_{50}$ of 1.11 ${\mu}M$ by SRB assay. To verify whether the cytotoxicity of Vitisin A may be associated with apoptosis, TdT-mediated-dUTP Nick-End Labeling (TUNEL) assay and cell cycle analysis were performed in MES-SA cells. Apoptotic bodies were observed in Vitisin A treated MES-SA cells by TUNEL assay. Also, Vitisin A effectively increased the portion of $sub-G_1$ DNA content by flow cytometric analysis. Taken together, these findings suggest that the cytotoxicity of Vitisin A against MES-SA cells is chiefly mediated by apoptosis.