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Relationship between Reactive Oxygen Species and Adenosine Monophosphate-activated Protein Kinase Signaling in Apoptosis Induction of Human Breast Adenocarcinoma MDA-MB-231 Cells by Ethanol Extract of Citrus unshiu Peel (진피 추출물에 의한 인간유방암 MDA-MB-231 세포의 apoptosis 유도에서 ROS 및 AMPK의 역할)

  • Kim, Min Yeong;HwangBo, Hyun;Ji, Seon Yeong;Hong, Su-Hyun;Choi, Sung Hyun;Kim, Sung Ok;Park, Cheol;Choi, Yung Hyun
    • Journal of Life Science
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    • v.29 no.4
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    • pp.410-420
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    • 2019
  • Citrus unshiu peel extracts possess a variety of beneficial effects, and studies on their anticancer activity have been reported. However, the exact mechanisms underlying this activity remain unclear. In the current study, the apoptotic effect of ethanol extract of C. unshiu peel (EECU) on human breast adenocarcinoma MDA-MB-231 cells and related mechanisms were investigated. The results showed that the survival rate of MDA-MB-231 cells treated with EECU was significantly inhibited in a concentration-dependent manner, which was associated with the induction of apoptosis. EECU-induced apoptosis was associated with the activation of caspase-8 and caspase-9, which initiate extrinsic and intrinsic apoptosis pathways, respectively, and caspase-3, a representative effect caspase. EECU suppressed the expression of the inhibitor of apoptosis family of proteins, leading to an increased Bax/Bcl-2 ratio and proteolytic degradation of poly (ADP-ribose) polymerase. EECU also enhanced the loss of the mitochondrial membrane potential and cytochrome c release from the mitochondria to the cytosol, along with truncation of Bid. In addition, EECU activated AMP-activated protein kinase (AMPK), and compound C, an AMPK inhibitor, significantly weakened EECU-induced apoptosis and cell viability reduction. Furthermore, EECU promoted the generation of reactive oxygen species (ROS), which acted as upstream signals for AMPK activation as pretreatment of cells, with the antioxidant N-acetyl cysteine reversing both EECU-induced AMPK activation and apoptosis. Collectively, these findings suggest that EECU inhibits MDA-MB-231 adenocarcinoma cell proliferation by activating intrinsic and extrinsic apoptotic pathways, which was mediated through ROS/AMPK-dependent pathways.

Molecular Characterization and Expression Analysis of Clathrin-Associated Adaptor Protein 3-δ Subunit 2 (AP3S2) in Chicken

  • Oh, Jae-Don;Bigirwa, Godfrey;Lee, Seokhyun;Song, Ki-Duk
    • Korean Journal of Poultry Science
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    • v.46 no.1
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    • pp.31-37
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    • 2019
  • A chicken clathrin-associated adaptor protein $3-{\delta}$ subunit 2 (AP3S2) is a subunit of AP3, which is involved in cargo protein trafficking to target membrane with clathrin-coated vesicles. AP3S2 may play a role in virus entry into host cells through clathrin-dependent endocytosis. AP3S2 is also known to participate in metabolic disease developments of progressions, such as liver fibrosis with hepatitis C virus infection and type 2 diabetes mellitus. Chicken AP3S2 (chAP3S2) gene was originally identified as one of the differentially expressed genes (DEGs) in chicken kidney which was fed with different calcium doses. This study aims to characterize the molecular characteristics, gene expression patterns, and transcriptional regulation of chAP3S2 in response to the stimulation of Toll-like receptor 3 (TLR3) to understand the involvement of chAP3S2 in metabolic disease in chicken. As a result, the structure prediction of chAP3S2 gene revealed that the gene is highly conserved among AP3S2 orthologs from other species. Evolutionarily, it was suggested that chAP3S2 is relatively closely related to zebrafish, and fairly far from mammal AP3S2. The transcriptional profile revealed that chAP3S2 gene was highly expressed in chicken lung and spleen tissues, and under the stimulation of poly (I:C), the chAP3S2 expression was down-regulated in DF-1 cells (P<0.05). However, the presence of the transcriptional inhibitors, BAY 11-7085 (Bay) as an inhibitor for nuclear factor ${\kappa}B$ ($NF{\kappa}B$) or Tanshinone IIA (Tan-II) as an inhibitor for activated protein 1 (AP-1), did not affect the expressional level of chAP3S2, suggesting that these transcription factors might be dispensable for TLR3 mediated repression. These results suggest that chAP3S2 gene may play a significant role against viral infection and be involved in TLR3 signaling pathway. Further study about the transcriptional regulation of chAP3S2 in TLR3 pathways and the mechanism of chAP3S2 upon virus entry shall be needed.

Inhibitory Effects of Prunus mume Solvent Fractions on Human Colon Cancer Cells (매실 분획물에 의한 인체 대장암세포 억제 효과)

  • Kim, Jeong-Ho;Cho, Hyun-Dong;Won, Yeong-Seon;Heo, Ji-An;Kim, Ji-Young;Kim, Hwi-Gon;Han, Sim-Hee;Moon, Kwang-Deog;Seo, Kwon-Il
    • Journal of Life Science
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    • v.29 no.11
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    • pp.1227-1234
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    • 2019
  • Prunus mume, also known as maesil, is a popular fruit consumed in East Asia (Korea, Japan, and China). It contains high amounts of organic acids, minerals, and polyphenols and has been used as a medication for fever, vomiting, and detoxification. In this study, the anti-proliferative and apoptotic effects of solvent fractions from maesil were evaluated using sulforhodamine B (SRB) assays, morphological evaluations, Hoechst 33258 staining, and western blotting. Addition of the maesil methanol fraction (MMF) and the maesil butanol fraction (MBF) significantly and dose-dependently decreased the cell viability of HT-29 human colon cancer cells. Colony-forming assays confirmed that the MMF and MBF treatments decreased colony numbers when compared with untreated control cells. Treatment of HT-29 cells with MMF and MBF caused a distortion of the cell morphology to a shrunken cell mass. Treatment with MMF and MBF also dose-dependently increased nuclear condensation and the formation of apoptotic bodies in HT-29 cells. Treatment with MMF and MBF significantly and dosedependently increased the expression of Bax (a pro-apoptotic protein), caspase-3, and poly ADP-ribose polymerase (PARP) and decreased the expression of Bcl-2 (an anti-apoptotic protein). MMF significantly and dose-dependently inhibited cell proliferation induced by bisphenol A, an environmental hormone. Therefore, MMF may have potential use as a functional food and as a possible therapeutic agent for the prevention of colon cancer.

Antioxidant and Anticancer Activities of Euonymus porphyreus Extract in Human Lung Cancer Cells A549 (인체 폐암 세포주 A549에서 Euonymus porphyreus 추출물의 항산화 및 항암활성 분석)

  • Jin, Soojung;Oh, You Na;Son, Yu Ri;Bae, Soobin;Park, Jung-ha;Kim, Byung Woo;Kwon, Hyun Ju
    • Journal of Life Science
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    • v.31 no.2
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    • pp.199-208
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    • 2021
  • Euonymus porphyreus, a species of plant in the Celastraceae family, is widely distributed in East Asia, especially in Southern China. The botanical characteristics of E. porphyreus have been reported, but its antioxidative and anticancer activities remain unclear. In this study, we evaluated the antioxidative and anticancer effects of ethanol extracts of E. porphyreus (EEEP) and the molecular mechanism of its anticancer activity in human lung adenocarcinoma A549 cells. The total polyphenol and flavonoid compound contents from EEEP were 115.42 mg/g and 23.07 mg/g, respectively. EEEP showed significant antioxidative effects with a concentration at 50% of the inhibition (IC50) value of 11.09 ㎍/ml, as measured by 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay. EEEP showed cytotoxic activity by increasing the SubG1 cell population of A549 cells in a dose-dependent manner. Apoptosis in A549 cells treated with EEEP was evident due to increased apoptotic cells and apoptotic bodies, as detected by Annexin V and 4,6-diamidino-2-phenylindole (DAPI) staining, respectively. EEEP-induced apoptosis resulted in increased expression of the First apoptosis signal (Fas), p53, and Bax, with decreased expression of Bcl-2 and subsequent activation of caspase-8, -9, and caspase-3, leading to cleavage of poly (ADP-ribose) polymerase (PARP). Collectively, these results suggest that EEEP may exert an anticancer effect by inducing apoptosis in A549 cells through both intrinsic and extrinsic pathways.

Effect of Cardanol Content on the Antibacterial Films Derived from Alginate-PVA Blended Matrix (알지네이트-폴리비닐알콜 블랜드 항균 필름 제조를 위한 카다놀 함량의 영향)

  • Ahn, Hee Ju;Kang, Kyung Soo;Song, Yun Ha;Lee, Da Hae;Kim, Mun Ho;Lee, Jae Kyoung;Woo, Hee Chul
    • Clean Technology
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    • v.28 no.1
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    • pp.24-31
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    • 2022
  • Petroleum-based plastics are used for various purposes and pose a significant threat to the earth's environment and ecosystem. Many efforts have been taken globally in different areas to find alternatives. As part of these efforts, this study manufactured alginate-based polyvinyl alcohol (PVA) blended films by casting from an aqueous solution prepared by mixing 10 wt% petroleum-based PVA with biodegradable, marine biomass-derived alginate. Glutaraldehyde was used as a cross-linking agent, and cardanol, an alkyl phenol-based bio-oil extracted from cashew nut shell, was added in the range of 0.1 to 2.0 wt% to grant antibacterial activity to the films. FTIR and TGA were performed to characterize the manufactured blended films, and the tensile strength, degree of swelling, and antibacterial activity were measured. Results obtained from the FTIR, TGA, and tensile strength test showed that alginate, the main component, was well distributed in the PVA by forming a matrix phase. The brittleness of alginate, a known weakness as a single component, and the low thermal durability of PVA were improved by cross-linking and hydrogen bonding of the functional groups between alginate and PVA. Addition of cardanol to the alginate-based PVA blend significantly improved the antibacterial activity against S. aureus and E. coli. The antibacterial performance was excellent with a death rate of 98% or higher for S. aureus and about 70% for E. coli at a contact time of 60 minutes. The optimal antibacterial activity of the alginate-PVA blended films was found with a cardanol content range between 0.1 to 0.5 wt%. These results show that cardanol-containing alginate-PVA blended films are suitable for use as various antibacterial materials, including as food packaging.

Effect on Physiological Metabolism of Calcium Ion at Cell Membrane Model of Parathyroid which Irradiated by High Energy X-ray (고에너지 엑스선을 조사한 부갑상선의 세포막모델에서 칼슘이온의 생리학적 대사에 미치는 영향)

  • Ko, In-Ho;Yeo, Jin-Dong
    • Journal of the Korean Society of Radiology
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    • v.16 no.2
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    • pp.141-150
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    • 2022
  • The initial co-transport and counter-transport permeate transport characteristics of calcium ion at epithelial cell membrane model in parathyroid which irradiated by high energy x-ray(linac 6 MV) was investigated. The epithelial cell membrane model used in this experiment was a polysulfonated copolymerized membrane of poly(PS-DVB: polystyrene-divinylbenzene). The difference of sorbed water in membrane, fixed carrier concentration(SO32-), initial pH value, OH- concentration were occurred at difference of Ca2+concentration and quantity of parathyroid hormone, respectively. The initial co-transport and counter-transport permeate flux of Cl-, OH-, Ca2+ on fixed carrier concentration(SO32-) and initial pH value of irradiated membrane was found to be decreased than non-irradiated membrane. The initial co-transport and counter-transport permeate flux of Ca2+ on fixed carrier concentration (SO32-), initial pH value, OH- concentration in irradiated membrane were found to be decreased about 2.68 ~ 6.87 times, about 1.42 ~ 1.63 times, about 2.07 ~ 1.672 times than non-irradiated membrane, respectively. As a result, the quantity of parathyroid hormone was decreased at irradiated membrane than non-irradiated membrane. The decrease of parathyroid hormone was occurred at hypoparathyroidism and osteoporosis, parathyroiditis, and so on. As the parathyroid hormone in epithelial cell membrane model were abnormal, cell damages were appeared at cell.

Apoptotic Effect of Proso Millet Grains on Human Breast Cancer MDA-MB-231Cells Is Exerted by Activation of BAK and BAX, and Mitochondrial Damage-mediated Caspase Cascade Activation (기장 종자 유래 추출물의 인간 유방암 MDA-MB-231 세포에 대한 세포독성에 관련된 미토콘드리아 손상-의존적 아폽토시스 유도 효과)

  • Do Youn Jun;Cho Rong Han;Young Ho Kim
    • Journal of Life Science
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    • v.33 no.1
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    • pp.15-24
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    • 2023
  • To examine the antitumor effect of proso millet grains, whether proso millet grains exert apoptotic activity against human cancer cells was investigated. When the cytotoxicity of 80% ethanol (EtOH) extract of proso millet grains was tested against various cancer cells using MTT assay, more potent cytotoxicity was observed against human breast cancer MDA-MB-231 cells than against other cancer cells. When the EtOH extract was evaporated to dryness, dissolved in water, and then further fractionated by sequential extraction using four organic solvents (n-hexane, methylene chloride, ethyl acetate, and n-butanol), the BuOH fraction exhibited the highest cytotoxicity against MDA-MB-231 cells. Along with the cytotoxicity, TUNEL-positive apoptotic nucleosomal DNA fragmentation and several apoptotic responses including BAK/BAX activation, mitochondria membrane potential (Δψm) loss, mitochondrial cytochrome c release into the cytosol, activation of caspase-8/-9/-3, and degradation of poly (ADP-ribose) polymerase (PARP) were detected. However, human normal mammary epithelial MCF-10A cells exhibited a significantly lesser extent of sensitivity compared to malignant MDA-MB-231 cells. Irrespective of Fas-associated death domain (FADD)-deficiency or caspase-8-deficiency, human T acute lymphoblastic leukemia Jurkat cells displayed similar sensitivities to the cytotoxicity of BuOH fraction, excluding an involvement of extrinsic apoptotic mechanism in the apoptosis induction. These results demonstrate that the cytotoxicity of BuOH fraction from proso millet grains against human breast cancer MDA-MB-231 cells is attributable to intrinsic apoptotic cell death resulting from BAK/BAX activation, and subsequent mediation of mitochondrial damage-dependent activation of caspase cascade.

Effects of Vanadate Solution Property on the Precipitation of Ammonium (Meta, Poly)Vanadate (바나데이트 수용액 특성이 암모늄(메타, 폴리)바나데이트 침전에 미치는 영향)

  • Ho-Sung Yoon;Seo Jin Heo;Yujin Park;Rina Kim;Chul-Joo Kim;Kyeong Woo Chung;Hong In Kim
    • Resources Recycling
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    • v.32 no.3
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    • pp.26-37
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    • 2023
  • Good control of the solution pH and temperature is required to recover vanadium from the water leaching solution of vanadium ore after sodium roasting. However, such adjustments could lead to aluminum-vanadium and sodium-vanadium co-precipitation, which greatly affects the efficiency of vanadium recovery. In this study, a process that can increase the efficiency of vanadium recovery as ammonium metavanadate [NH4VO3] and ammonium polyvanadate [(NH4)2V6O16·H2O] was investigated by examining the characteristics of vanadium-containing aqueous solutions during precipitation. The aluminum content of vanadium-containing water leaching solutions has a great effect on the loss of vanadium when the pH of the aqueous solution is adjusted to 9. Therefore, a process to minimize aluminum leaching is also required. In this study, ~99% or more of vanadium present in vanadium-containing aqueous solutions was precipitated and recovered as NH4VO3 by adding 3 equivalents of ammonium chloride relative to the vanadium content at pH 9 and room temperature. (NH4)2V6O16·H2O was precipitated from the aluminum-vanadium coprecipitates generated during the pH-adjustment of the aqueous solutions to 9 by dissolving the coprecipitate in the solutions at pH 2.5 and controlling their sodium content to 2,000 mg/L or less. Approximately, 98% or more of the available (NH4)2V6O16·H2O could be precipitated and recovered from a solution with a vanadium content of 2,200 mg/L and a sodium content of 1,875 mg/L at pH 2.5 by adding approximately 3 equivalents of ammonium chloride relative to the vanadium content at 95℃ or higher. The overall process could precipitate and recover, approximately 91% or more of the total vanadium in the water leaching solution as NH4VO3 and (NH4)2V6O16·H2O.

Isoalantolactone Inhibits the Formation of Multicellular Tumor Spheroids Derived From Human Hepatocellular Carcinoma Hep3B Cells Through the Induction of ROS-dependent Apoptosis (ROS 의존적 세포사멸 유도를 통한 isoalantolactone의 인간 간세포암종 Hep3B 세포 유래 다세포 종양 spheroid 형성의 억제)

  • Min Yeong Kim;Byunwoo Son;Sang-Hyup Lee;Sang Eun Park;Su Hyun Hong;Sang Hoon Hong;Eunjeong Kim;Yung Hyun Choi;Hyun Hwangbo
    • Journal of Life Science
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    • v.34 no.7
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    • pp.476-484
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    • 2024
  • Although two-dimensional (2D) monolayer cell culture models are still widely used as the optimal models for anticancer activity research, three-dimensional (3D) multicellular tumor spheroid (3D MTS) models that can better approximate the tumor environment can offer an alternative to bridge the gap between in vitro and animal model studies. Isoalantolactone is among the sesquiterpene lactones found in medicinal plants, including the roots of Elecampane (Inula helenium L.), and is known to have various pharmacological activities, including anticancer activity. In this study, we investigated whether the anticancer activity of isoalantolactone observed in 2D models could be reproduced in a 3D MTS model derived from human hepatocellular carcinoma (HCC) Hep3B cells. According to our results, isoalantolactone inhibited the formation of MTSs in a manner dependent on the treatment concentration, which was accompanied by an increase in reactive oxygen species (ROS) generation. In particular, as isoalantolactone treatment and the culture time increased, the area of proliferating cells was replaced by cells in which apoptosis was induced. Additionally, in MTSs, isoalantolactone increased the expression of death-receptor-related proteins and the activity of caspase-3, and it decreased the expression of the Bax/Bcl-2 expression ratio and total poly(ADP-ribose) polymerase. However, when the production of ROS was artificially blocked, all these changes caused by isoalantolactone were attenuated and the cell survival rate of MTS cells was restored. Therefore, the results of this study suggest that the induction of apoptosis in Hep3B cell-derived MTSs by isoalantolactone is achieved through the activation of extrinsic and intrinsic pathways and is ROS-dependent.

Larvae Growth and Biochemical Composition Change of the Pacific Oyster Crassostra gigas, Larvae during Artificial Seed Production (참굴 Crassostrea gigas 인공종묘생산 시 유생의 성장과 체성분 변화)

  • Hur, Young-Baek;Min, Kwang-Sik;Kim, Tae-Eic;Lee, Seung-Ju;Hur, Sung-Bum
    • Journal of Aquaculture
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    • v.21 no.4
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    • pp.203-212
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    • 2008
  • A nutritional demand of oyster, Crassostrea gigas larva as part of research for improving of utilization of microalgae being used for the artificial oyster seed production. The change of body growth and biochemical compositions of larvae were investigated during larvae rearing in hatchery. The larvae were cultured in 60 M/T tank and fed mixture 6 different phytoplankton species, Isochrysis galbana (30%), Cheatoceros gracilis (20%), Pavlova lutheri (20%), Phaeodactylum triconutum (10%), Nannochryis oculata (10%) and Tetraselmis tetrathele (10%). The initial feeding amount was $0.3{\times}10^4cells/mL$ at three times a day to D-shaped larva and the feeding amount had been increased 30% gradually every two day since the larvae were raising. The larvae were developed from D shape to pediveliger stage for 12 days. The daily growth of shell length and hight were $5.8{\sim}30.8\;{\mu}m$ and $8.7{\sim}31.4\;{\mu}m$, respectively and weight gains were changed from D shape to pediveliger as follow: wet weight was $0.52{\sim}15.0\;{\mu}g/larva$, dry weight was $0.2{\sim}6.5\;{\mu}g/larva$, and ash free dry weight was $0.1{\sim}8.5\;{\mu}g/larva$. The larvae growth pattern shown a logarithmic phase from D shape to umbone stage but after that stage shown a exponential growth aspect. The crude protein, crude lipid and nitrogen free extract (NFE) of larvae during rearing periods were analyzed as $6.1{\sim}10.6%$, $0.6{\sim}1.1%$ and 1.0-2.7%, respectively. And the total amino acid contents of the larvae during rearing periods were in order as glutamic acid $1.26{\sim}2.24%$, aspartic acid $0.97{\sim}1.70%$, and methionine $0.12{\sim}0.33%$. Of the total fatty acid in the analyzed larvae, the saturated fatty acid (SSAFA) was decreased from 54.3% (D shaped larvae) to 17.1 % (pediveliger) as larvae development but the total mono-unsaturated fatty acid (${\Sigma}MOFA$) and Poly-unsaturated fatty acid (${\Sigma}PUFA$) were increased from 29.9% and 7.8% to 40.6% and 45.6%, respectively. By the way the each fatty acid of the larvae were composed of palmitic acid $9.89{\sim}36.95%$, oleic acid $12.17{\sim}32.29%$, linoleic acid $1.96{\sim}33.55%$, EPA $2.17{\sim}11.58%$ and DHA $1.95{\sim}4.51%$. As a result of this study, the larvae of oyster were demanded a various nutrients for healthy growth and the feeding control, expecially after umbone stage larvae are a rapidly growing time, is very important for success of artificial seed production.