• 제목/요약/키워드: Cytotoxic protein

검색결과 471건 처리시간 0.037초

Discovery of LDD-1075 as a potent FLT3 inhibitor

  • Kyoung Bin Yoon;Hyo Jeong Lee;Hye Jin Chung;Jungeun Lee;Jungil Choi;Jeong Doo Heo;Yong‑Chul Kim;Sun‑Young Han
    • Oncology Letters
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    • 제17권5호
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    • pp.4735-4741
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    • 2019
  • Fms-like tyrosine kinase 3 (FLT3) is a valuable pharmacological target in the treatment of acute myeloid leukemia (AML). LDD-1075 and LDD-1076 are indirubin derivatives, and LDD-1075 is the ester form of LDD-1076. LDD-1076 exhibited a potent in vitro FLT3 kinase activity inhibition with an IC50 of 7.89 nM, whereas, LDD-1075 demonstrated a relatively weak activity against FLT3 (IC50 of 3.19 µM). In contrast with the results of the FLT3 kinase activity inhibition assay, the LDD-1076 did not affect the growth of the MV4-11 cell line, which harbors the constitutively activated form of the FLT3 mutation. Notably, LDD-1075 exhibited a strong cytotoxic effect against the MV4-11 cells. When LDD-1075 was incubated with the MV4-11 cell lysate, the formation of LDD-1076 was observed. Treatment with LDD-1075 inhibited the FLT3 phosphorylation along with the phosphorylation of the signal transducer and activator of transcription 5 protein, which is a downstream signal transducer of FLT3. Treatment with LDD-1075 induced apoptosis and cell cycle arrest at the G1 phase. The present study demonstrated that the LDD-1076 formed by the bioconversion of LDD-1075 is a potent FLT3 inhibitor with anti-leukemic activity.

참가사리 분획물의 암 예방효과 (Anticarcinogenic Effects of Extracts from Gloiopeltis tenax)

  • 정영화;정복미;신미옥;배송자
    • 한국식품영양과학회지
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    • 제35권4호
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    • pp.395-401
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    • 2006
  • 본 연구에서는 해조류 중 홍조류에 속하는 참가사리를 추출, 분획하여 항발암효과를 측정하였다. 참가사리 분획물을 4종의 암세포주 HT29, HepG2, MCF-7 및 H16-F10에 처리하였을 때 암세포 증식 억제실험을 한 결과 대장암세포주인 HT29의 경우 GTMM층과 GTMB층에서 농도의존적인 효과가 나타났으며 첨가농도 $150{\mu}g/mL$에서 각각 93.64%와 74.14%의 수치를 보였다. 간암세포주인 HepG2의 경우 $150{\mu}g/mL$ 첨가시 GTMM층과 GTMB층이 각각 95.97%, 81.78%의 높은 증식 억제효과를 보였으며 유방암세포주인 MCF-7에서는 GTMM층이 $120 {\mu}g/mL$$150{\mu}g/mL$ 첨가농도에서 각각 88.50%와 91.82%의 높은 암세포증식 억제효과를 나타냈다. 피부암세포주인 B16-F10은 다른 세포주에 비해 미약하나 역시 GTMM층이 최종첨가농도에서 86.20%의 억제를 나타내었다. 이와 같이 4종의 암세포주의 증식억제는 전반적으로 GTMM층과 GTMB층에서 높은 효과가 나타남을 알 수 있다. 한편 Western blot analysis를 통해 간암세포 주인 HepG2에서 GTMM층의 처리에 따른 pro-apoptosis Bcl-2 family의 발현증가를 볼 수 있었으며, PARP 분절과 연관된 caspase-3, 7의 활성화를 확인할 수 있었다. 이처럼 GTMM층은 apoptosis의 작용에 의한 세포사멸을 유도하며, 이러한 apoptosis의 기전으로는 최소한 caspase의 활성화에 따른 절단 PARP 단백질의 증가, Bad, Bax 등의 apoptosis 관여인자가 작용하고 있음을 확인할 수 있었다. HepG2를 이용한 quinone reductase 유도활성여부를 측정한 결과 GTMM층이 $45{\mu}g/mL$$60{\mu}g/mL$의 시료첨가농도에서 대조군에 비해 각각 2.34, 2.86배의 QR 유도활성을 나타내었으며, GTMB층의 경우 최종첨가농도인 $60{\mu}g/mL$에서 2.04배의 효소활성을 나타내었다.

LPS로 유도한 RAW 264.7 세포의 염증반응에서 흰민들레의 항염증 효과 (The anti-inflammatory effect of Taraxacum coreanum on lipopolysaccharide induced inflammatory response on RAW 264.7 cells)

  • 김민준;배기상;최선복;조일주;김동구;신준연;이성곤;김명진;박성주;송호준
    • 대한본초학회지
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    • 제29권6호
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    • pp.21-26
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    • 2014
  • Objectives : Taraxacum coreanum (TC) have been used as a traditional medicine to treat inflammatory diseases and anti-oxidant effect in Korea. However, the anti-inflammatory effect of TC water extract on lipopolysaccharide (LPS)-induced inflammation is not well-known. Therefore, this study was performed to identify the anti-inflammatory effect of TC on LPS induced inflammatory. Methods : RAW 264.7 cells were treated with 500 ng/mL of LPS. Water extracts of TC (0.1, 0.25, 0.5 mg/ml) was treated 1 h prior to LPS. Cell viability was measured by MTT assay. Levels of nitric oxide (NO) were measured with Griess reagent and pro-inflammatory cytokines were measured by enzyme-linked immunosorbent assay (ELISA) and real-time polymerase chain reaction (real-time PCR). We also examined molecular mechanisms such as mitogen-activated protein kinases (MAPKs) and nuclear factor-B ($NF-{\kappa}B$) activation by western blot. Results : Water Extract from TC itself did not have any cytotoxic effect in RAW 264.7 cells. TC treatment inhibited the production of NO production, and pro-inflamamtory cytokines such as interleukin (IL)-6 and $IL-1{\beta}$ on protein and mRNA levels. In addition, TC treatment inhibited the LPS-induced activation of MAPKs such as extracellular signal-regulated kinase1/2 (ERK1/2), p38 kinases (p38), c-Jun $NH_2$-terminal kinase (JNK) and $NF-{\kappa}B$. Conclusions : In summary, our result suggest that treatment of TC could reduce the LPS-induced inflammation. Thereby, TC could be used as a protective agent against inflammation. Also, this study could give a clinical basis that TC could be a drug or agent to prevent inflammation.

LPS로 유도된 RAW 264.7 세포와 마우스 귀 조직에 대한 참도박(Grateloupia elliptica Holmes) 에탄올 추출물의 항염증 효과 (Anti-Inflammatory Effect of Ethanol Extract from Grateloupia elliptica Holmes on Lipopolysaccharide-Induced Inflammatory Responses in RAW 264.7 Cells and Mice Ears)

  • 배난영;김민지;김꽃봉우리;안나경;최연욱;박지혜;박선희;안동현
    • 한국식품영양과학회지
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    • 제44권8호
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    • pp.1128-1136
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    • 2015
  • 본 연구에서는 참도박 에탄올 추출물(GEHEE)의 항염증 효과를 알아보기 위해 lipopolysaccharide에 의해 활성화된 대식세포로부터 분비되는 염증매개인자들의 발현량과 마우스 모델을 이용한 귀 부종 및 조직학적 관찰 실험을 진행하였다. 그 결과 염증을 유발하는 대표적 물질인 NO 및 사이토카인[interleukin(IL)-6, $IL-1{\beta}$ 및 tumor necrosis factor $receptor-{\alpha}$]의 분비량이 GEHEE 농도 의존적으로 유의적 감소를 보였다. 또한 전사인자인 nuclear $factor-{\kappa}B$의 활성과 인산화효소인 mitogen-activated protein kinases(p38, JNK 및 ERK)의 발현량이 억제됨을 확인함에 따라 염증작용 기전에서 이들의 활성 억제가 NO 및 사이토카인 분비량 조절에 영향을 미침을 확인하였다. 동물모델을 이용한 실험에서는 croton oil로 부종을 유발한 마우스 귀 조직에 GEHEE를 처리하였을 때 항염증제인 prednisolone 처리구와 유사한 수준으로 경피 및 진피의 두께가 감소한 것을 확인하였으며, 조직 내 침윤되는 mast cell의 수도 현저히 억제됨을 확인하였다. 따라서 본 연구 결과는 참도박 에탄올 추출물의 항염증 효능을 가진 기능성 소재로의 이용 가능성을 제시한다.

재래감귤 팔삭의 과피 추출물이 LPS로 활성화 된 RAW264.7 대식세포에서 염증매개물질 억제에 미치는 효과 (Inhibition of LPS-induced Inflammatory Biomarkers by Fraction of Citrus hassaku pericarp through Suppression of NF-${\kappa}B$ Activation in RAW264.7 Cells)

  • 김철원;김성무;정승원;김소미;안광석
    • 대한암한의학회지
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    • 제16권2호
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    • pp.25-34
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    • 2011
  • Objectives : Citrus is the fruit that is readily available around us. Therefore, we investigated the anti-inflammatory effects of fraction isolated from the Citrus hassaku pericarp in RAW264.7 macrophage cells. Methods : The effects of fraction from Citrus hassaku pericarp on cell viability on RAW264.7 cells were measured by the MTT assay. The mRNA levels of iNOS and COX-2, its protein level by fraction of Citrus hassaku pericarp treatment in RAW264.7 macrophage cells were investigated by RT-PCR and immunoblots. Nitrite accumulation in the culture was measured colorimetrically by the Griess reaction using a Griess reagent. The amount of IL-6 and TNF-${\alpha}$ production was determined using an enzyme-linked immunosorbent assay (ELISA) kit. Results : The results indicated that the fraction of Citrus hassaku pericarp concentration highly suppressed lipopolysaccharide (LPS)-induced nitric oxide (NO) and IL-6 productions without a cytotoxic effect on RAW264.7 cells. fraction of Citrus hassaku pericarp inhibited the expressions of LPS-induced iNOS and COX-2 protein and their mRNA in a dose-dependent manner. Particularly, fraction of Citrus hassaku pericarp suppressed the level of nuclear factor-${\kappa}B$ (NF-${\kappa}B$) activity, which was linked with the suppression of LPS-induced phosphorylation of p65 at serine 276 and p65 translocation into nuclei, but not MAPK signaling. In addition, treatment with fraction of Citrus hassaku pericarp inhibited the production of IL-6 and TNF-${\alpha}$ in LPS-stimulated RAW264.7 cells. Conclusion : Our results indicate that fraction of Citrus hassaku pericarp potentially inhibits the biomarkers related to inflammation through the blocking of NF-${\kappa}B$ p65 activation, and it may be a potential therapeutic candidate for the treatment of inflammatory diseases.

생체유리와 천연산호 골이식재가 치주인대 섬유아세포 활성에 미치는 영향 (Biological Effects of bioactive glass and natural coral on periodontal ligament fibroblast-like cell behavior)

  • 심성규;한수부
    • Journal of Periodontal and Implant Science
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    • 제29권1호
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    • pp.173-192
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    • 1999
  • The purpose of this study was to evaluate the effects of bioactive glass and natural coral on the human periodontal ligament fibroblast(HPLF) behaviors during the regeneration process of peridontium. To determine the cellular events occuring in the presence of the particles of bioactive glass and natural coral, HPLF were isolated from healthy premolar teeth extracted for orthodontic treatment. Cells were cultured in ${\alpha}$MEM at 37$^{\circ}C$, 5% $CO_2$, 95% humidity incubator. Bioactive glass and natural coral were powdered, and each particles(<40${\mu}$m) were placed on the cultured cells at the concentration of 0.3mg/ml, and 1,0mg/ml for experimental group. In control group no particles were added. And each group was evaluated by examining the cell morphology under phase-contrast micrograph at 4 day and transmission electron micrograph(TEM) and scanning electron micrograph(SEM) at 14 day, alkaline phosphatase activity at 5 and 9 day, protain synthesis at 4 day, DNA synthesis at 1, 2, 3 and 4 day, cell proliferation at 1, 3, 5,7 and 9 day and the formation of bone nodule at 30 day after culturing all groups in mineralizing supplemented mediun, No significant changes in cell morphology by adding these two matirials were found under phase contrast microscopy and TEM. HPLF phagocytocized each particles suggesting that HPLF is involved in the process of resorbing each particles and that bioactive glass were more biocompatible than natural coral. The ALPase activity of bioactive glass 0.3 mg/ml was similar with control groups and all the rests of control groups were significantly low(P<0.01) indicating a transient dedifferentiation of HPLF in the presence of bioactive glass and natural coral particles. There were no significant differences of protein synthesis between all groups. The DNA synthesis in experimental groups were significantly lower than control groups at 1, 2 and 3 day (P<0.01) but became similar to control groups at 4 day. Between control groups, the DNA synthesis in bioactive glass O.3mglml group was significantly higher than other groups(P<0.01). Cell proliferation in natural coral 1.0mg/ml and bioactive glass 1.0mglml groups were significantly lower than control group at 3 day(P<0.05) and there were no differences at 5, 7, 9 day. There were more bone nodule formation in experimental groups than in control groups. In conclusion, these results indicated that bioactive glass and natural coral have some effects of a transient dedifferentiation on HPLF and regeneration of periodontal tissues, however any significant cytotoxic effect on HPLF by these two particles were not found.

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반응표면분석에 의한 해송이버섯(Hypsizigus marmoreus) 추출물 중 단백다당체의 암세포 성장억제효과 (Cytotoxic Effect of Isolated Protein-bound Polysaccharides from Hypsizigus marmoreus Extracts by Response Surface Methodology)

  • 정은봉;조진호;조승목
    • 한국식품영양과학회지
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    • 제37권12호
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    • pp.1647-1653
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    • 2008
  • 해송이버섯 분말의 암세포성장억제효과의 증진을 위한 성분의 추출조건을 최적화하기 위하여 반응표면분석법을 사용하였다. 중심합성계획에 따라 추출조건의 독립변수(추출온도, 추출시간, 용매비)와 이에 따라 영향을 받는 수율과 추출물의 단백질 함량을 종속변수로 설정하였다. 수율과 단백질 함량이 높은 조건을 충족시키는 최적조건은 추출온도 $51.3^{\circ}C$, 추출시간 8.2시간, 그리고 추출용매비는 46.7 mL/g 으로 나타났다. 최적조건에서 추출되어진 조추출물을 이용하여 알코올을 이용하여 침전한 후 dialysis tube를 이용하여 저분자물질을 제거하여 조다당체를 얻었으며 조다당체를 이용하여 이온교환수지를 이용하여 산성다당체와 중성다당 체를 얻어 각각의 다당체의 암세포성장억제효과를 알아보았다. 위암세포주 AGS에는 산성다당체가 0.5 mg/mL의 농도에서 73.97%로 가장 높은 세포증식억제효과를 보였으며 간암세포주 HepG2에는 조다당체가 0.5 mg/mL의 농도에서 82.45%로 가장 높은 효과를 보였다. 그러나 결장암세포주 SW480에서는 모든 분획의 시료가 20%미만의 세포성장 억제효과를 보였다.

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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    • 제30권2호
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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 $PPAR{\alpha}$.

부자(附子)와 오가피(五加皮) 물 추출물의 골수유래 지방세포와 파골세포 분화 억제 효과 (Inhibitory Effect of Water Extracts of Aconiti Lateralis Preparata Radix and Acanthopanacis Cortex on Differentiation of Bone Marrow-Derived Adipocytes and Osteoclasts)

  • 이경선;최은식;한상용;김윤경
    • 대한한의학방제학회지
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    • 제22권1호
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    • pp.151-165
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    • 2014
  • Objectives : The aim of this study was to evaluate the efficacy of Aconiti Lateralis Preparata Radix (AP) and Acanthopanacis Cortex (AT) extracts in bone-derived adipocyte OP9 cell, osteoclast and osteoblast-like MG63 cells. Methods : MTT assay was used to evaluate the cytotoxicity of AP and AT extracts on OP9, osteoclast and MG63 cells. OP9 cells were treated with AP and AT, and the alterations in fat storage in the cells were determined by the Oil red O. To explain effects of RANKL-induced osteoclast differentiation in bone marrow macrophages, we performed the TRAP staining. The protein level of CAAAT/enhancer binding protein alpha ($C/EBP{\alpha}$) and peroxisome proliferator-activated receptor ${\gamma}$ ($PPAR{\gamma}$) as a adipocyte differentiation marker, and adiponectin was examined using western blot in differentiated OP9 cells. Effects of related genes were confirmed by luciferase assay using reporter assay. Results : AP and AT was not toxic on OP9 and MG63 cells, but AT was a little cytotoxic to osteoclast at the dose of $100{\mu}g/m{\ell}$. They could inhibit differentiation of OP9 cells and osteoclast with results of oil red O staining and TRAP staining. By western blot, AP and AT decreased the expression of $PPAR{\gamma}$ and $C/EBP{\alpha}$ which is the key transcription factor in adipogenesis and adiponectin secretion. AT also inhibited the BMP-4 activity in luciferase assay. AP also inhibited BMP-4 and Wnt3a activity, stimulated ER-${\beta}$ activity but inhibited androgen receptor activity. Conclusions : These results show AP and AT can be useful in osteoporosis and obesity via inhibition of osteoclast and adipocyte differentiation.

상기생과 봉독이 간암 세포주 Hep G2에 대해 미치는 항암 기전 비교 (Comparative Study of Korean Mistletoe Lectin and Bee Venom on the Anti-Cancer Effect and Its Mechanisms of Action in Hepatocellular Carcinoma Cells)

  • 김승욱;김보람;허경;임성우
    • 대한한방내과학회지
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    • 제30권4호
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    • pp.845-857
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    • 2009
  • Background and Objectives : Korean mistletoe lectin (Viscum album coloratum agglutinin, VCA) and bee venom (BV) have been reported to induce apoptosis in various cancer cell lines in vitro and to show antitumor activity against a variety of tumors in animal models. However, the comparative effect of VCA and BV on the anti-cancer effect and mechanisms of action has not been determined. In this study, the effect in a human hepatocellular carcinoma cell line, Hep G2 cells, was examined. Methods : Cytotoxic effects of VCA and BV on Hep G2 cells were determined by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay in litro. The apoptotic cell death was then confirmed by propidium iodide staining and DNA fragmentation analysis. The mechanisms of action were examined by the expression of anti-apoptotic proteins and activation of mitogen-activated protein kinases. The involvement of kinase was examined in VCA or BV-induced apoptosis by using kinase inhibitors. Results : VCA and BV killed Hep G2 cells in a time and dose-dependent manner. Treatment of Hep G2 cells with VCA activated poly (ADP-ribose) polymerase-1 (PARP-1) known as a marker of apoptosis, and mitogen-activated protein kinases signaling pathways including MAPK/ERK, p38 MAPK and JNK. BV also activated PARP-1, MAPK/ERK. and p38 MAPK but not JNK. The expression level of anti-apoptotic molecule, Bcl-X, was decreased by VCA treatment but not by BV. Finally, the phosphorylation level of ERM proteins involved in the cytoskeleton homeostasis was decreased by both stimuli. VCA-induced apoptosis was partially inhibited by in the presence of JNK and p38 inhibitor, but BV only by p38 inhibitor. Conclusions : VCA-induced apoptosis is dependent on the activation of p38 and JNK. while BV-induced apoptosis is mediated by p38 activation in Hep G2 cells.

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