• Title/Summary/Keyword: 억제인자

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Induction of Apoptosis by β-Lapachone in Hep3B Human Hepatocellular Carcinoma Cells Is Caspase-Dependent and Associated with Inactivation of PI3K/Akt Signaling (Hep3B 인간 간암세포에서 caspase 의존적이며 PI3K/Akt 신호전달의 불활성화와 관련된 β-lapachone의 세포사멸 유도)

  • Jae Im Kwon;Yung Hyun Choi;Hyun Hwangbo
    • Journal of Life Science
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    • v.34 no.2
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    • pp.94-104
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    • 2024
  • β-Lapachone is a natural quinone compound originally obtained from the bark of the lapacho tree (Tabebuia vellanedae), which has been used in traditional medicine in several South and Central American countries for treating various diseases. Although β-lapachone has been reported to have potent anticancer activity in many types of cancer cells, its effect on the proliferation of hepatocellular carcinoma (HCC) cells is still unclear. Therefore, in this study, we investigated the effect of β-lapachone on the proliferation of human HCC Hep3B cells. According to our results, the decrease in cell viability of Hep3B cells caused by β-lapachone was closely related to the induction of apoptosis, which was confirmed through changes in nuclear morphology and flow cytometry. In addition, in Hep3B cells treated with β-lapachone, the expression of Bcl-2, an anti-apoptotic factor, was decreased, while the expression of Bax, an apoptosis inducer, was increased, and the activity of the caspase cascade was also increased. However, in the presence of a pan-caspase inhibitor, β-lapachone-induced apoptosis was weakened, indicating that the induction of apoptosis by β-lapachone was caspase-dependent. Moreover, β-lapachone treatment activated extracellular-regulated kinase (ERK) signaling while inhibiting activation of the phosphoinositide 3 kinase (PI3K)/Akt pathway. Furthermore, the effect of the ERK inhibitor on suppressing the induction of apoptosis by β-lapachone was minimal, and the PI3K inhibitor significantly increased β-lapachone-induced apoptosis. The findings from this study will contribute to a better understanding of the anticancer activity of β-lapachone in HCC cells.

Anti-inflammatory effects of biorenovated Torreya nucifera extract in RAW264.7 cells induced by Cutibacterium acnes (여드름균에 의해 유도된 RAW264.7 세포에서 생물 전환된 비자나무 추출물의 항염증 효과)

  • Hyehyun Hong;Tae-Jin Park;Yu-Jung Lee;Byeong Min Choi;Seung-Young Kim
    • Journal of Applied Biological Chemistry
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    • v.66
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    • pp.213-220
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    • 2023
  • The most common skin disease, acne, often occurs in adolescence, but it is also detected/observed in adults due to air pollution and drug abuse. One of the causative agents of acne, Cutibacterium acnes (C. acnes) plays a role in the development of skin acne by inducing inflammatory mediators. Torreya nucifera (TN) is an evergreen tree of the family Taxaceae, having well reported antioxidant, anti-proliferative, liver protection, and nerve protection properties. Improvement of these bioactive properties of natural products is one of the purposes of natural product chemistry and pharmaceuticals. We believe biorenovation could be one improvement strategy that utilizes microbial metabolism to produce unique derivatives having enhanced bioactivity. Therefore, in this study, the C. acnes-induced RAW264.7 inflammation model was used to evaluate the anti-inflammatory activity of the biorenovated Torreya nucifera product (TNB). The results showed improved viability of TNB-treated cells compared to TN-treated cells in the concentration range of 50, 100, and 200 ㎍/mL. At non-toxic concentrations, TNB inhibited the production of nitric oxide and prostaglandin E2 by suppression of inducible nitric oxide synthase and cyclooxygenase-2 protein expression. TNB also attenuated the expression of interleukin-1β, interleukin-6, interleukin-8, and tumor necrosis factor-α induced by C. acnes. Furthermore, TNB inhibited the nuclear factor-κB signaling pathway, a transcription factor known to regulate inflammatory mediators. Based on these results, this study suggests the potential of using TNB as natural material for the treatment of acnes and thus, supporting our postulation of biorenovation as an bioactivity improvement strategy.

AtMYB7 Acts as a repressor of lignin biosynthesis in Arabidopsis (애기장대 MYB7 유전자의 리그닌 생합성 억제 조절)

  • Kim, Won-Chan
    • Journal of Applied Biological Chemistry
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    • v.59 no.3
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    • pp.215-220
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    • 2016
  • Abstract Secondary cell wall is the most abundant biomass produced by plants. Plant secondary cell wall is composed of a complex mixture of cellulose, hemicellulose, and lignin. Lignin, a phenolic polymer that hinders the degradation of cell wall polysaccharides to simple sugars destined for fermentation to bio-ethanol. Cell wall biosynthesis pathway-specific biomass engineering offers an attractive 'genetic pretreatment' strategy to improve bioenergy feedstock. Recently, we found a transcription factor, MYB7, which is a transcriptional switch that may turns off the genes necessary for lignin biosynthesis. To gain insights into MYB7 mediated transcriptional regulation, we first established a dominant suppression system in Arabidopsis by expressing MYB7-SRDX. Then we used a transient transcriptional activation assay to confirm that MYB7 suppress the transcription of the lignin biosynthetic gene. Taken together, we conclude that MYB7 function as a repressor of the genes involved in the lignin biosynthesis.

Inhibitory Effects of Artemisia fukudo Makino Extracts for Nitric Oxide Generation in LPS- and Interferon-${\gamma}$-stimulated RAW 264.7 Cells (갯쑥(Artemisia fukudo Makino) 추출물의 LPS와 Interferon-${\gamma}$로 자극한 RAW264.7 세포주의 NO 생성 저해 효과)

  • Lee, Joon-Kyoung;Oh, Sung-Hoon
    • Journal of the East Asian Society of Dietary Life
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    • v.18 no.2
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    • pp.198-206
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    • 2008
  • 갯쑥(Artemisia fukudo Makino, 일명 바다쑥)은 바닷가 염전 주변 습지에서 자라는 염생식물로 어린 갯쑥은 나물로 먹어왔으며, 민간에서 간에 좋다고 알려져 있다. 본 연구에서는 염생식물의 기능 성분을 탐색하는 연구의 일환으로 마우스유래 대식세포주인 RAW264.7세포를 이용하여 갯쑥의 nitric oxide(NO) 생성 억제 작용을 알아보고자 하였다. 갯쑥은 methanol 추출과 열수 추출한 후 농축하여 실험하였고, methanol 추출물은 다시 hexane, chloroform, ethylacetate, butanol, water로 순차 용매 분획하여 농축하여, 각각의 추출물을 RAW 264.7 세포에 농도별 분획별로 처리한 후 LPS와 interferon-${\gamma}$로 자극한 RAW 264.7 세 포주의 NO 생성을 측정하여 NO 생성 저해율을 구하였다. 갯쑥 methanol 추출물의 높은 NO 생성 저해율과 iNOS 활성 저해율을 나타내었으며(IR=90.7%과 IR=81.6%), 각각의 추출물중 분획물이 100${\mu}g/mL$ 농도에서 hexane 분획물이 가장 높은 NO 생성 저해 효과(IR=92.1%)와 iNOS 활성 억제 효과(IR=85.0%)를 나타내었다. 갯쑥의 methanol 추출물, hexane과 chloroform 분획물에서 높은 NO 생성 저해 효과를 나타내어 새로운 암 예방인자 물질로 기대되며, 따라서 본 연구 결과는 암 예방인자 분리를 위한 기초 자료가 될 것이며, 나아가 성분 분리 연구와 동물 실험을 통한 기전 연구가 수행되어야 할 것이다.

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The Effect of Needle Electrode Electrical Stimulation Following Global Ischemia on the Suppression of Apoptosis in the Cerebellum (전뇌허혈 유발 후 침전극저주파자극 적용이 소뇌의 세포자멸사 억제에 미치는 영향)

  • Wang, Joong-San;Park, Joo-Hyun;Moon, Ok-Kon;Kim, Nyeon-Jun;Choi, Jung-Hyun;Kim, Ji-Sung
    • The Journal of the Korea institute of electronic communication sciences
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    • v.8 no.12
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    • pp.1949-1958
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    • 2013
  • This study conducted observation of tissues using the immunohistochemistry methods in order to see the effect of Needle Electrode Electrical Stimulation(: NEES) on apoptosis in the SD rat cerebellum caused by global ischemia which came from common carotid artery occlusion. The comparison and analysis results of expression of factors related to apoptosis among the control group, the global ischemia group and the NEES group showed that NEES following global ischemia had a significant effect on the reduction of c-fos expression in the SD rat cerebellum, and didn't have a significant effect on the reduction of bax and caspase-3 expression. In conclusion, it is considered that NEES following global ischemia is partially effective in suppressing the expression of factors related to apoptosis.

Antioxidative Activities of Skipjack Meat Extract (가다랑어 자숙엑스분의 항산화성)

  • OH Kwang-Soo;LEE Eung-Ho;KIM Myung-Chan;LEE Kang-Hee
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.20 no.5
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    • pp.441-446
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    • 1987
  • To develop cheaper and safer natural antioxidative substance instead of synthesized ones, the extract of skipjack meat was examined for the antioxidative effects and factors. Amino-N of skipjack meat extract was 15.3 mg/100g, and the extract apparently showed inhibitory effect on the oxidation of methyl linoleate. Antioxidative activity of extract revealed a tendancy to reduce slightly in proportion to hydrolyzing. When extract was dialyzed in distilled water, the outer fraction had a strong inhibitory effect on the oxidation of nethyl linoleate, while the inner fraction showed no effect. From the omission test and chemical analysis, the major antioxidative factors of skipjack meat extract were amino acids such as anserine, histidine, carnosine and alanine. As well nucleotides such as AMP, hypoxanthine seemed to act an auxiliary role in antioxidative effect of that.

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Inhibition of cellular proliferation and apoptosis by Scutellaria Baicalensis in MDA-MB-231 breast cancer cells (황금의 유방암세포주에 대한 항암작용)

  • Yong, Hyung-Soon;Ko, Seong-Gyu
    • The Journal of Internal Korean Medicine
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    • v.25 no.3
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    • pp.451-460
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    • 2004
  • Objective : 황금의 유방암세포주에 대한 항암효과 및 기전에 대한 연구는 아직 미흡하며, 특히 에스트로젠리셉터를 가지지않은 유방암세포주인 MDA-MB-231에 대한 효과 및 기전에 대한 연구는 아직 발표된바 없어, 이에 대한 연구가 진행되었다. Methods : 인간 유방암세포주 MDA-MB-231 MTT assay를 이용 성장방해비율을 조사하였으며. FACS analysis를 이용 cell cycle analysis를 시행하였고, Western Blot Analysis 및 Annexin V analysis를 시행하였다. Results : MDA-MB-231에 대한 황금의 IC50는 180 ug/ml 이었으며 최대 세포성장억제효과는 $500{\mu}g/ml$로 한약재중 비교적 강한 세포독성을 보여 주었다. 유세포분석 에서 황금 $500{\mu}g/ml$의 농도를 72시간 투여한 경우 세포사멸(Sub Gl) 분율이 대조군의 1.7%에 비해 21%로 높아 현저한 용량의존적인 세포사멸현상을 보여주었으며, 세포사멸을 보다 명확히 규명할 수 있는 Annexin V analysis에서도 황금 $200{\mu}g/ml$농도일때 48시간에서 17%의 뚜렷한 세포사멸효과를 나타내었다. 한편 세포사멸촉진인자인 Bax, 세포사멸실행단백질인자인 caspase 3의 활성과 PARP의 분할은 세포사멸이 세포주기정지와 더불어 세포사멸의 과정에 p53이 관여함을 알 수 있다. 앞으로의 연구는 p53발현이 다른 세포주와 각 단백질의 억제제를 통해 인과적인 관련성을 즘 더 명확히 할 필요가 있어야 할 것으로 생각되어진다. Conclusion : 유방암의 예후에 있어 호르몬치료에 부적절함으로 인해 예후가 나쁜 에스트로젠리셉터 발현이 없는 유방암에 대해서도 황금이 탁월한 항암효과를 보여주고 있으며, 임상적으로 황금단독, 다른 항암약재와의 배합, 그리고 기존의 항암화학요법이나 방사선요법과의 병용투여를 통한 초기 및 진행된 유방암의 치료에 대한 새로운 접근의 실마리를 제공할 것으로 생각된다.

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Expression of rpr, grim, dcp-1, diapl, and diap2 during Drosophila Development (노랑초파리 발생과정에서 rpr, grim, dcp-1, diapl, diap2의 발현)

  • Park, Ji-Gweon;Chung, Ki-Wha;Kim, Se-Jae
    • Development and Reproduction
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    • v.5 no.2
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    • pp.131-136
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    • 2001
  • The developmental profiles of rpr, grim, dcp-1, diapl, diap2 transcripts, which were involved in programmed cell death, were analyzed using competitive RT-PCR in whole animals during Drosophila development. The fluctuation patterns of transcript levels of the apoptotic initiators(rpr and grim) were similar to those of the ecdysone titer in Drosophila life cycle. The transcript of dcp-1, which is considered as effector caspase, was expressed strong1y at early embryo and female adult stages. However, the transcript levels of anti-apoptotic factors diap1 and diap2, showed the reverse pattern comparing with those of apoptotic factors(rpr and grim). Also, the transcript levels of rpr, diap2 and dcp-1 were quantified in the salivary glands and wing discs dissected from the wandering late third instar larva. The transcript levels of rpr and diap2 were changed reversely each other in both tissues from wandering stage to puparium formation. These results suggest that the expressions of cell death related genes are regulated by the ecdysone signals during normal development.

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Establishment of Reporter Cell Lines that Monitor Activities of Hypoxia Inducible Factor-1, P53 and Nur77 for Assessment of Carcinogenicity (저산소유도인자 HIF-1, 암 억제인자 P53과 고아 핵수용체 Nur77의 발현을 지표로 하는 발암독성예측 세포주의 개발)

  • Hong, Il;Seo, Hee-Won;Lee, Min-Ho;Kim, Ji-Won;Chung, Jin-Ho;Lee, Byung-Hoon;Lee, Mi-Ock
    • Toxicological Research
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    • v.23 no.3
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    • pp.231-238
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    • 2007
  • Evaluation of potentials of chemicals to alter expression of genes that are involved in carcinogenesis may serve useful tools in toxicological research. In this investigation, we developed reporter cell lines that expressed luciferase in response to transactivation of hypoxia inducible factor-1, P53 tumor suppressor and Nur77 of which roles have been well established in cancer development and progression. Whereas these reporter cell lines displayed low constitutive backgrounds, the reporter activities were significantly enhanced in response to $desferriosamine/CoCl_2$, adriamycin or 6-mercaptopurine, which are hypoxia mimicking chemicals, P53 activator or Nur77 inducer, respectively. The activation of the reporter was time- and dose-dependent. Known tumor initiators and promoters, such as phorbol 12-myristate 13-acetate and phorbol 12, 13-dicaprinate induced the reporter activity at as low as 10nM in these stable cell lines. Further, known anti-tumor promoters, such as ascorbic acid and ${\beta}-carotene$ repressed the reporter activities. These results indicate that our stable reporter cell lines could serve as a useful system for rapid assessment of carcinogenicity of toxic chemicals.

Anti-Inflammatory Activities of Hog Millet (Panicum miliaceum L.) in Murine Macrophages through IRAK-4 Signaling (대식세포에서 IRAK-4신호조절을 통한 기장(Panicum miliaceum L.)의 항염증능에 관한 연구)

  • Park, Mi-Young;Kim, Jae-Hyun;Park, Dong-Sik
    • The Korean Journal of Food And Nutrition
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    • v.24 no.2
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    • pp.268-272
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    • 2011
  • 전곡류의 섭취와 만성질환의 유병율은 음의 상관관계가 있는 것으로 알려져 있다. 본 연구에서는 선행 연구 결과, 지방축적억제능이 우수한 소재로 선정된 기장의 항염증능 여부를 검증하고자 하였다. 이를 위해 RAW264.7 세포에 기장열수분획($1\;{\mu}g/m\ell$$10\;{\mu}g/m\ell$)과 lipopolysaccharide(LPS)를 함께 처리한 후, 24시간 배양시켜 염증매개인자들의 분비량 및 mRNA 발현 정도를 측정하였다. 또한 LPS 자극에 대한 첫 번째 신호전달인자로 알려져 있는 interleukin-1 receptor associated kinase-4(IRAK-4)의 단백질 발현 정도를 측정하였다. 본 연구결과, 기장의 열수분획($10\;{\mu}g/m\ell$)은 LPS로 유도된 NO, $PGE_2$, TNF-${\alpha}$, IL-6 및 MCP-1의 생성량 및 mRNA 발현량을 유의적으로 억제하였다(p<0.05). 특히 이들 지표 중 pro-inflammatory cytokine인 TNF-${\alpha}$와 IL-6의 mRNA 발현량이 효과적으로 감소하였다(p<0.01). IRAK-4의 단백질 발현량 또한 유의적으로 감소하여 LPS 자극에 대한 기장열수분획의 항염증능은 toll-like receptor(TLR)를 통한 IRAK-4를 매개로 하는 신호전달체계 조절에 기인하는 것으로 사료된다.