• 제목/요약/키워드: MTT reduction

검색결과 223건 처리시간 0.033초

LPS로 활성화된 RAW 264.7 cell에서 하경(荷梗)의 염증매개물질 억제효과 (Anti-inflammtory effects of the MeOH extract of Petiolus Nelumbinis)

  • 이원욱;조미정;박상미;정지윤;김상찬
    • 대한한의학방제학회지
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    • 제17권1호
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    • pp.175-185
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    • 2009
  • Petiolus Nelumbinis, branches of lotus leaf or lotus flower is a traditional oriental herbal medicine widely used for treating a superheat or disorder of qi flow. Although there are many clinical results and literature study, it has been rarely conducted to evaluate the immuno-biological activity. The present study was conducted to examine the anti-inflammatory effects of PNM (Petiolus Nelumbinis MeOH extract) in vitro. To determine cytotoxic concentration of PNM, the cells were treated with PNM for 24 h after LPS addition, and the cell viability was tested by MTT assay. Both of dosages (30 and 100 ${\mu}g/ml$) of PNM had no cytotoxicity. In these concentrations, PNM significantly reduced the elevated levels of NO and $PGE_2$ by LPS. These inhibitory effects of PNM were due to the reduced expressions of iNOS and COX-2 protein. TNF-$\alpha$, IL-1$\beta$ and IL-6 are frequently encountered pro-inflammatory cytokines, and LPS plays a key role in inducing to the massive production of these cytokines. Thus, we next determined the levels of these cytokines. Although PNM had no significant inhibitory effect on the production of TNF-$\alpha$, the elevated levels of IL-1$\beta$ and IL-6 by LPS were dose-dependently reduced in PNM-treated groups. These results demonstrate that PNM has anti-inflammatory effects by inhibiting the production of proinflammatory cytokines, NO and $PGE_2$ in LPS-activated macrophage. Moreover, the reduction of NO and $PGE_2$ levels was due to the inhibition of iNOS and COX-2 protein expression by PNM.

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VVC의 기울기 기반 화면내 예측모드 결정 및 블록분할 고속화 기법 (Gradient-Based Methods of Fast Intra Mode Decision and Block Partitioning in VVC)

  • 윤용욱;박도현;김재곤
    • 방송공학회논문지
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    • 제25권3호
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    • pp.338-345
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    • 2020
  • 차세대 비디오 부호화 표준으로 진행중인 VVC(Versatile Video Coding)는 HEVC(High Efficiency Video Coding)보다 두 배 이상의 압축 성능을 달성하기 위해 다양한 기술들을 채택하고 있다. 최근 배포된 VVC 참조 SW 코덱인 VTM(VVC Test Model)은 HEVC 대비 38% 이상의 BD-rate 부호화 성능 향상을 보이는 반면 부호화와 복호화 복잡도가 각각 9배, 2배 정도 증가를 보인다. 특히, 재귀적 MTT(Multi-Type Tree) 분할 구조와 HEVC 대비 2배로 증가한 화면내 예측모드 수로 인해 상당한 부호화기의 복잡도가 증가하였으며, 이를 감소시키기 위한 다양한 기법들이 연구되고 있다. 본 논문에서는 부호화기의 복잡도를 감소시키기 위하여 블록내 화소의 기울기를 이용한 고속 화면내 예측모드 결정 및 블록분할 기법을 제시한다. 실험결과 VTM6.0 대비 AI(All Intra) 부호화 구조에서 3.54%의 부호화 성능 감소와 65%의 부호화 시간 절감 효과를 얻었다.

Ameliorating Effect of Gardenia jasminoides Extract on Amyloid Beta Peptide-induced Neuronal Cell Deficit

  • Choi, Soo Jung;Kim, Mi-Jeong;Heo, Ho Jin;Hong, Bumshik;Cho, Hong Yon;Kim, Young Jun;Kim, Hye Kyung;Lim, Seung-Taik;Jun, Woo Jin;Kim, Eun-Ki;Shin, Dong-Hoon
    • Molecules and Cells
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    • 제24권1호
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    • pp.113-118
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    • 2007
  • The brains of Alzheimer's disease (AD) patients are characterized by large deposits of amyloid beta peptide ($A{\beta}$). $A{\beta}$ is known to increase free radical production in nerve cells, leading to cell death that is characterized by lipid peroxidation, free radical formation, protein oxidation, and DNA/RNA oxidation. In this study, we selected an extract of Gardenia jasminoides by screening, and investigated its ameliorating effects on $A{\beta}$-induced oxidative stress using PC12 cells. The effects of the extract were evaluated using the 2',7'-dichlorofluorescein diacetate (DCF-DA) assay and the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction assay. To find the active component, the ethanol extract was partitioned with hexane, chloroform, and ethyl acetate, respectively, and the active component was purified by silica-gel column chromatography and HPLC. The results suggested that Gardenia jasminoides extract can reduce the cytotoxicity of $A{\beta}$ in PC 12 cells, possibly by reducing oxidative stress.

Blocking Bcl-2 Leads to Autophagy Activation and Cell Death of the HEPG2 Liver Cancer Cell Line

  • Du, Peng;Cao, Hua;Wu, Hao-Rong;Zhu, Bao-Song;Wang, Hao-Wei;Gu, Chun-Wei;Xing, Chun-Gen;Chen, Wei
    • Asian Pacific Journal of Cancer Prevention
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    • 제14권10호
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    • pp.5849-5854
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    • 2013
  • Background: Apoptosis may be induced after Bcl-2 expression is inhibited in proliferative cancer cells. This study focused on the effect of autophagy activation by ABT737 on anti-tumor effects of epirubicin. Methods: Cytotoxic effects of ABT737 on the HepG2 liver cancer cell line were assessed by MTT assay and cell apoptosis through flow cytometry. Mitochondrial membrane potential was measured by fluorescence microscopy. Monodansylcadaverin (MDC) staining was used to detect activation of autophagy. Expression of p53, p62, LC3, and Beclin1, apoptotic or autophagy related proteins, was detected by Western blotting. Results: ABT737 and epirubicin induced growth inhibition in HepG2 cells in a dose- and time-dependent manner. Both ABT737 and epirubicin alone could induce cell apoptosis with a reduction in mitochondrial membrane potential as well as increased apoptotic protein expression. Further increase of apoptosis was detected when HepG2 cells were co-treated with ABT373 and epirubicin. Furthermore, our results demonstrated that ABT373 or epirubicin ccould activate cell autophagy with elevated autophagosome formation, increased expression of autophagy related proteins and LC3 fluorescent puncta. Conclusions: ABT737 influences cancer cells through both apoptotic and autophagic mechanisms, and ABT737 may enhance the effects of epirubicin on HepG2 cells by activating autophagy and inducing apoptosis.

The Effects of Purified Artemia Extract Proteins on Proliferation, Differentiation and Apoptosis of Human Leukemic HL-60 Cells

  • Deezagi, Abdolkhaleg;Chashnidel, Azadeh;Hagh, Neda Vaseli;Shahraki, Mahvash Khodabandeh
    • Asian Pacific Journal of Cancer Prevention
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    • 제17권12호
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    • pp.5139-5145
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    • 2016
  • There has been an increment in the number of studies focused on marine bioactive materials. Many peptides and other biomaterials with anticancer potential have been extracted from various marine animals. Artemia extracts have found uses in sun-light protection cosmetics and anti-aging products. However, contents of biochemical compounds in Artemia spp. and molecular mechanisms of have not been clearly studied in leukemic cells in vitro. In this work, we isolated and purified proteins of Artemia Urmiana. Six clear fractions (A-F) observed on DEAE-cellulose chromatography were assayed for effects on cell growth, differentiation and apoptosis using the human leukemic HL-60 cell line. Cell proliferation analysis by MTT and BrdU assays indicated that did not affect cells, growth. Cells treated with crude extract and fractions A, B and C, but not E and F (up to $100{\mu}g/mL$), exhibited increase of cell growth in a dose dependent manner. Stimulatory effects of fraction D were observed at concentrations of $10{\mu}g/mL$ and above. In nitro blue tetrazolium (NBT) reduction assays, treatment with $100{\mu}g/mL$ of fraction E or F for 96 hr increased the fraction of differentiated cells up to $14.8{\pm}3.56%$ and $16.5{\pm}2.08%$ respectively. Combination of those fractions with retinoic acid had significant synergistic effects on the differentiation of cells ($56.8{\pm}3.7%$ and $67.4{\pm}4.2%$, $p{\leq}0.01$). Annexin-V FITC staining for apoptosis and flow cytometric assays indicated induction of apoptosis by fractions E and F up to 23.8 and 31.8% of cells.

Radical Intermediate Generation and Cell Cycle Arrest by an Aqueous Extract of Thunbergia Laurifolia Linn. in Human Breast Cancer Cells

  • Jetawattana, Suwimol;Boonsirichai, Kanokporn;Charoen, Savapong;Martin, Sean M
    • Asian Pacific Journal of Cancer Prevention
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    • 제16권10호
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    • pp.4357-4361
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    • 2015
  • Thunbergia Laurifolia Linn. (TL) is one of the most familiar plants in Thai traditional medicine that is used to treat various conditions, including cancer. However, the antitumor activity of TL or its constituents has never been reported at the molecular level to support the folklore claim. The present study was designed to investigate the antitumor effect of an aqueous extract of TL in human breast cancer cells and the possible mechanism(s) of action. An aqueous crude extract was prepared from dried leaves of TL. Folin-Ciocalteu colorimetric assays were used to determine the total phenolic content. Antiproliferative and cell cycle effects were evaluated in human breast adenocarcinoma MCF-7 cells by MTT reduction assay, cell growth inhibition, clonogenic cell survival, and flow cytometric analysis. Free radical generation by the extracts was detected using electron paramagnetic resonance spectroscopy. The exposure of human breast adenocarcinoma MCF-7 cells to a TL aqueous extract resulted in decreases in cell growth, clonogenic cell survival, and cell viability in a concentration-dependent manner with an $IC_{50}$ value of $843{\mu}g/ml$. Treatments with extract for 24h at $250{\mu}g/ml$ or higher induced cell cycle arrest as indicated by a significant increase of cell population in the G1 phase and a significant decrease in the S phase of the cell cycle. The capability of the aqueous extract to generate radical intermediates was observed at both high pH and near-neutral pH conditions. The findings suggest the antitumor bioactivities of TL against selected breast cancer cells may be due to induction of a G1 cell cycle arrest. Cytotoxicity and cell cycle perturbation that are associated with a high concentration of the extract could be in part explained by the total phenolic contents in the extract and the capacity to generate radical intermediates to modulate cellular proliferative signals.

Short Low Concentration Cisplatin Treatment Leads to an Epithelial Mesenchymal Transition-like Response in DU145 Prostate Cancer Cells

  • Liu, Yi-Qing;Zhang, Guo-An;Zhang, Bing-Chang;Wang, Yong;Liu, Zheng;Jiao, Yu-Lian;Liu, Ning;Zhao, Yue-Ran
    • Asian Pacific Journal of Cancer Prevention
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    • 제16권3호
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    • pp.1025-1028
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    • 2015
  • Background: Prostate cancer is one of the main causes of cancer death, and drug resistance is the leading reason for therapy failure. However, how this occurs is largely unknown. We therrfore aimed to study the response of DU145 cells to cisplatin. Materials and Methods: Du145 prostate cancer cells were treated with a low dose of cisplatin for 24 h and cell viability and number were determined by MTT assay and trypan blue exclusion assay, respectively. The real time polymerase chain reaction (PCR) was used to assess responses to cisplatin treatment. Results: After 24h $2{\mu}g/ml$ treatment did not result in significant reduction in cell viability or number. However, it led to enhanced cancer cell invasiveness. E-cadherin mRNA was reduced, and vimentin, Snail, Slug, metalloproteinase 9 (MMP9) mRNA expression increased significantly, a feature of epithelial-mesenchymal transition (EMT). Conclusions: Short time low concentration cisplatin treatment leads to elevated invasiveness of DU145 cancer cells and this is possibly due to EMT.

인체 간암세포에서 비기환(肥氣丸), 대칠기탕(大七氣湯) 및 목향빈랑환(木香檳榔丸) 열수 추출물의 항암 활성 비교 (Comparison of Anti-cancer Potentials of Water Extracts of Bigihwan, Daechilgithang and Mokwhyangbinranghwan in Human Hepatocellular Carcinoma Cells)

  • 김민영;이혜숙;홍수현;박철;최영현
    • 대한한의학방제학회지
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    • 제28권1호
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    • pp.15-27
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    • 2020
  • Objectives : We selected three herb-combined remedies, Bigihwan (BGH), Daechilgitang (DCGT) and Mokwhyangbinranghwan (MHBRH), through Donguibogam text-mining analysis, and evaluated their anti-cancer effects on human hepatocellular carcinoma Hep3B cells. Methods : Cytotoxicity was assessed by an MTT assay. Apoptosis rate, autophagy and ROS level were detected by flow cytometry. The autophagy was also observed by Cyto-ID staining fluorescence microscopy. The expression of autophagy, mitophagy and pexophagy regulatory proteins was detected by Western blot analysis. Results : BGH showed the strongest effect among the three prescriptions in inhibiting Hep3B cell viability, which was associated with the induction of apoptosis and autophagy. Autophagy blockers improved cell viability and reduced apoptosis after BGH and DCGT treatments, indicating that autophagy by these prescriptions enhanced Hep3B cells against their cytotoxicity. However, MHBRH enhanced the reduction of cell viability and apoptosis by autophagy blockers. Induction of autophagy by BGH treatment was associated with mitophagy due to mitochondrial dysfunction than DCGT and MHBRH-treated cells. In addition, induction of apoptosis by BGH treatment was ROS-dependent and showed the possibility of pexophagy involvement. Conclusion : Although further studies need to be conducted to study the efficacy and mechanism of accurate anticancer activity, the present results will serve as important sources of understanding the mechanism of action of herbal remedies prescribed for liver disease as documented in Donguibogam.

Inhibition of Growth and Induction of Differentiation of SMMC-7721 Human Hepatocellular Carcinoma Cells by Oncostatin M

  • Kong, N.;Zhang, X.M.;Wang, H.T.;Mu, X.P.;Han, H.Z.;Yan, W.Q.
    • Asian Pacific Journal of Cancer Prevention
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    • 제14권2호
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    • pp.747-752
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    • 2013
  • Oncostatin M (OSM) is a multifunctional cellular regulator acting on a wide variety of cells, which has potential roles in the regulation of gene activation, cell survival, proliferation and differentiation. Previous studies have shown that OSM can induce morphological and/or functional differentiation and maturation of many tumor cells. However, the action of OSM on the induction of differentiation of human hepatocellular carcinoma (HCC) has not been reported. Here, we investigated the effects of different concentrations of OSM on human HCC cell line SMMC-7721 growth, proliferation, cell cycling, apoptosis and differentiation in vitro. Cell growth was determined via MTT assay, proliferation by cell cycle analysis, apoptosis by flow cytometry, morphology by transmission electronic microscopy, and cell function by detection of biochemical markers. Our results demonstrated that OSM strongly inhibited the growth of SMMC-7721 cells in a dose-dependent manner, associated with decreased clonogenicity. Cell cycle analysis revealed a decreased proportion of cells in S phase, with arrest at G0/G1. The apotosis rate was increased after OSM treatment compared to the control. These changes were associated with striking changes in cellular morphology, toward a more mature hepatic phenotype, accompanied by significant reduction of the expression of AFP and specific activity of ${\gamma}$-GT, with remarkable increase in secretion of albumin and ALP activity. Taken together, our findings indicate that OSM could induce the differentiation and reduce cell viability of SMMC-7721 cells, suggesting that differentiation therapy with OSM offers the opportunity for therapeutic intervention in HCC.

Antioxidant Effects of Gamma-oryzanol on Human Prostate Cancer Cells

  • Klongpityapong, Papavadee;Supabphol, Roongtawan;Supabphol, Athikom
    • Asian Pacific Journal of Cancer Prevention
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    • 제14권9호
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    • pp.5421-5425
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    • 2013
  • Background: To assess the antioxidant effects of gamma-oryzanol on human prostate cancer cells. Materials and Methods: Cytotoxic activity of gamma-oryzanol on human DU145 and PC3 prostate cancer cells was determined by proliferation assay using 3-(4, 5-dimethylthiazol, 2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) reagent. mRNA levels of genes involved in the intracellular antioxidant system, superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX) and glutathione reductase (GSR) were determined by reverse transcription-polymerase chain reaction (RT-PCR). Cancer cell lysates were used to measure lipid peroxidation using thiobarbituric acid reactive substance (TBARS). Glutathione contents of the cell lysates were estimated by the reaction between sulfhydryl group of 5, 5'-dithio (bis) nitrobenzoic acid (DTNB) to produce a yellow-color of 5-thio-2-nitrobenzoic acid using colorimetric assay. Catalase activity was also analysed by examining peroxidative function. Protein concentration was estimated by Bradford's assay. Results: All concentrations of gamma-oryzanol, 0.1-2.0mg/ml, significantly inhibited cell growth in a dose- and time-dependent fashion in both prostate cancer cell lines, DU145 and PC3. Gene expression of catalase in DU145 and PC3 exposed to gamma-orizanol at 0.5mg/ml for 14 days was down regulated, while mRNA of GPX was also down regulated in PC3. The MDA and glutathione levels including catalase activity in the cell lysates of DU145 and PC3 treated with gamma-oryzanol 0.1 and 0.5mg/ml were generally decreased. Conclusions: This study highlighted effects of gamma-oryzanol via the down-regulation of antioxidant genes, catalase and GPX, not cytotoxic roles. This might be interesting for adjuvant chemotherapy to make prostate cancer cells more sensitive to free radicals. It might be useful for the reduction of cytotoxic agents and cancer chemoprevention.