• Title/Summary/Keyword: MTT cell viability assay

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ABT-737 ameliorates docetaxel resistance in triple negative breast cancer cell line

  • Hwang, Eunjoo;Hwang, Seong-Hye;Kim, Jongjin;Park, Jin Hyun;Oh, Sohee;Kim, Young A;Hwang, Ki-Tae
    • Annals of Surgical Treatment and Research
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    • v.95 no.5
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    • pp.240-248
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    • 2018
  • Purpose: This study aimed to validate the synergistic effect of ABT-737 on docetaxel using MDA-MB-231, a triple negative breast cancer (TNBC) cell line overexpressing B-cell lymphoma-2 (Bcl-2). Methods: Western blot analysis was performed to assess expression levels of Bcl-2 family proteins and caspase-related molecules. Cell viability was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Cell cycle distribution was determined by flow cytometry analysis. Benzyloxycarbonyl-Val-Ala-Asp(OMe)-fluoromethylketone (z-VAD-fmk) was used for pretreatment to assess the role of caspases. Results: Cell viability of MDA-MB-231 after combination treatment with ABT-737 and docetaxel was significantly lower than that after docetaxel or ABT-737 monotherapy based on MTT assay (both P < 0.001), with a combination index of 0.41. The proportion of sub-G1 population after combination treatment was significantly higher than that after docetaxel or ABT-737 monotherapy (P = 0.001, P = 0.003, respectively). Pretreatment with z-VAD-fmk completely restored cell viability of MDA-MB-231 from apoptotic cell death induced by combination therapy (P = 0.001). Although pro-caspase-8 or Bid did not show significant change in expression level, pro-casepase-9 showed significantly decreased expression after combination treatment. Cleaved caspase-3 showed increased expression while poly (ADP-ribose) polymerase cleavage was induced after combination treatment. However, hypoxia-inducible factor 1-alpha and aldehyde dehydrogenase 1 totally lost their expression after combination treatment. Conclusion: Combination of ABT-737 with docetaxel elicits synergistic therapeutic effect on MDA-MB-231, a TNBC cell line overexpressing Bcl-2, mainly by activating the intrinsic pathway of apoptosis. Therefore, adjunct of ABT-737 to docetaxel might be a new therapeutic option to overcome docetaxel resistance of TNBCs overexpressing Bcl-2.

Effect of Omija (Schizandra Chinensis Baillon) Extracts on the Growth of Liver Cancer Cell Line SNU-398 (오미자 추출물의 간암세포 (SNU-398) 증식 억제 효과)

  • 노숙령;오현석
    • Journal of Nutrition and Health
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    • v.35 no.2
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    • pp.201-206
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    • 2002
  • This study was designed to research the anti-tumor effect of omija (methanol extract(I), malic acid & ethanol extract(II), and water extract (III)) on human liver cancer cell line SNU-398. MTT assay was used in vitro. The longer th\ulcorner exposure time and the higher the concentration of Omija extract, the stronger the anti-tumor effect. When the concentration of (II) was 1,600 $\mu\textrm{g}$/$m\ell$ and the exposure time reached 96 hours, The strongest propagation inhibition effect occurred with the viability rate as low as 5.06%. $IC_{50}$/ value was 363 $\mu\textrm{g}$/$m\ell$. Under the condition of 1,600$\mu\textrm{g}$/$m\ell$ and 96 hours, (I) lowered the rate to 7.75%. $IC_{50}$/ value was 489 $\mu\textrm{g}$/$m\ell$. When it was 1,600$\mu\textrm{g}$/$m\ell$ and 72 hours, (III) the rate decreased to 15.97%. $IC_{50}$/ value was 703 $\mu\textrm{g}$/$m\ell$. In all three cases, the viability of the cancer cell decreased significantly when the exposure time ranged between 24 and 48 hours.

Protective effects of added Bo-Yang-Hwan-Oh-Tang on $H_2O_2-induced$ neurotoxicity in SH-SY5Y neuronal cells (가매보양환오탕(加昧補陽還五湯)의 SH-SY5Y 뇌신경세포에서 산화적 손상에 의한 세포사멸에 대한 보호효과)

  • Han, Hyung-Soo;Park, Yong-Ki
    • The Korea Journal of Herbology
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    • v.21 no.4
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    • pp.85-92
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    • 2006
  • Objectives : To evaluate the neuroprotective effects of added Bo-Yang-Hwan-Oh-Tang (BHT), we investigated the neuronal death protection effects to oxidative damages in SH-SY5Y neuronal cells. Methods : To study the cytotoxic effects of BHT on SH-SY5Y cells, the cell viability was determined by MTT assay. To investigate the neuronal death protection of BHT, SH-SY5Y cells were induced oxidative damages by $H_2O_2$ and then assayed the cell viability and DNA fragmentation. We also investigated DPPH free radical scavenging effect of BHT by tube test. Results : In MTT assay, $1000{\mu}g/ml$ of BHT was not showed the cytotoxic effect on SH-SY5Y cells. BHT protected SHSY5Y cells from $H_2O_2-induced $ neuronal cell death in a dose-dependent manner. BHT also protected SH-SY5Y cells from $H_2O_2-induced$ DNA fragmentation. BHT effectively scavenged DPPH free radicals in a dose-dependent manner. Conclusion : These data suggest that BHT may have strong antioxidant effects through the free radical scavenging and neuroprotective effects in human neuronal cells.

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Antioxidant Activity and Protective Effects of Extracts from Sambucus williamsii var. coreana on t-BHP Induced Oxidative Stress in Chang cells (접골목 추출물에 의한 항산화 활성이 정상 간세포의 t-BHP 유발 산화스트레스에 미치는 영향)

  • Kim, Kitae
    • The Journal of the Society of Korean Medicine Diagnostics
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    • v.17 no.3
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    • pp.275-286
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    • 2013
  • In the present study, antioxidant activity and protective effect of extracts from Sambucus williamsii var. coreana stems (SWC) were evaluated on tert-butyl hydroperoxide (t-BHP) induced oxidative stress in human liver (Chang) cells. Antioxidant activities of the SWC extracts were determined by various radical scavenging activities, such as DPPH, ferric reducing antioxidant power (FRAP), 2,2'-azinobis-(3-ethybenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging activity and oxygen radical absorbance capacity (ORAC) assay. SWC extracts showed strong antioxidant effect on various assay. To determine the hepatoprotective effects of SWC on t-BHP induced oxidative damage, cell viability was measured using MTT assay. Pretreatment of SWC extracts showed increasing cell viability, decreasing ROS and restoring mitochondria membrane potential on t-BHP induced oxidative stress in Chang cells. Our findings suggest that SWC extracts may be considered a potential agent for therapeutic protective effect from oxidative stress through its antioxidant activity.

Cytotoxicity(MTT) evaluation of dental instruments made of polymers (치과용 폴리머 기구의 세포독성(MTT) 평가)

  • Choi, Eun-Mi
    • Journal of the Korea Convergence Society
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    • v.12 no.8
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    • pp.187-195
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    • 2021
  • In order to assess the cell toxicity of 10 instruments made of polymers, the MTT assay which utilizes the L-929 cell was selected. Specimens were eluted at a temperature of 37℃ for 24 hours at a rate of 4g per 20mL, RPMI 1640, and then was positively and negatively contrasted with a control test solution, in accordance with the Notification No. 2020-12 Protocols of Medical Apparatus Biological Safety from the Ministry of Drug and Food Safety. As a result of 24 hours of incubation in 37℃, 5% CO2 Incubator and assessment using an ELISA reader, the results of Intraoral camera indiciated a cellular viability of more than 70% at a 50% eluate. But, the Plastic impression tray, 3D printing tweezer, Impression disposable syringe, Dental floss holder, Hand implant scaler, Surgical retractor, Oral scanner tip, Dental mirror, and the Water pick tip all reported a cellular viability of more than 70% at a 100% eluate, which indicates that do not exhibit cytotoxicity, thus allowing it to be used in contact with the mucous membrane of the oral cavity.

Effects of Jangwon-hwan(Zhuangyuan-wan) Water Extract against Xanthine Oxidase / Hypoxanthine-induced Neurotoxicity in the Cultured Mouse Cerebral Cortical Neurons (장원환이 XO/HX에 의해 손상된 대뇌피질 신경세포에 미치는 영향)

  • 김영수;권강범;민영기;조현익;박준배;이호섭;류도곤
    • The Journal of Korean Medicine
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    • v.20 no.4
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    • pp.3-10
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    • 2000
  • In order to elucidate the toxic mechanism of neurotoxical damage and neuroprotective effect of Jangwon-hwan(Zhuangyuan-wan) water extract, this experiment was performed. Neurotoxic effects of xanthine oxidase/hypoxanthine(XO/HX) were examined by MTT and NR assay, neuroprotective effects of Jangwon-hwan(Zhuangyuan-wan) water extract were examined by neurofilament enzymeimmuno assay(EIA). XO/HX induced an increase in cell viability, and a decrease in the amount of neurofilament on cultured mouse cerebral cortical neurons in dose-dependent manner. In neuroprotective effect of herb medicine, Jangwon-hwan(Zhuangyuan-wan) water extract increased the amount of neurofilament on cultured mouse cerebral cortical neurons damaged by XO/HX. From the results, it is suggested that XO/HX showed toxic effect in cultured mouse cerebral cortical Neurons and Jangwon-hwan(Zhuangyuan-wan) water extract is very effective in the prevention of neurotoxicity induced by XO/HX.

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A Cytotoxicity of Carrier Oil and Essential Oils on Cells by Using of Aromatherapy (향기요법에 사용하는 캐리어 오일과 에션셜 오일의 세포에 대한 독성)

  • Yu, Byong-Soo;Kim, Sha-Sha;Yun, Young-Han;Kim, Ki-Young
    • Korean Journal of Human Ecology
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    • v.17 no.5
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    • pp.1027-1035
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    • 2008
  • Essentail oils and carrier oils are generally used for Aromatherapy. Therefore the toxicity, possibilities of irritations and sensitive reactions and injury of essential oils must be considered for clients and therapists. So that, in this studies a toxicity of jojoba and 4 species essential oils (fennel, mandarine, tea tree and cedarwood) were investigated by the measurement of MTT-assay and sirius red staining. Liver, kidney and brain tell were chosen for the cell viability assay and observation of morphological change. In the result, no cytotoxicity was observed on live., kidney and brain cell at concentration of 0.01 $\mu\el/m\el$ jojoba oil. And lysis and nucleus breaking were not observed at same concentration of jojoba oil on live., kidney and brain cell. fennel oil was showed 50% of cell viability and inhibited cell growth on liver, kidney and brain cell at relatively high concentration compared with the other oils. 50% of liver, kidney and brain cell viability and delayed cell growth of tea tree and mandarine oil were revealed at lower concentration than fennel oil. In cedarwood oil, 50% of liver cell viability at concentration of 0.00067 $\mu\el/m\el$ was showed, but cell viability and cell growth of kidney and beam cell were effected at the lowest concentration compared with other oils. So that, jojoba oil as using of carrier oil may be not harmful. And 3 essential oils from the fennel, tea tree and mamdarine may have very low toxicity, but cedarwood may be used carefully for inhalation. And over dosage of concentrated cedarwood oil should be not directly touched and exposured, and absolute essential oils must be diluted with carrier oils for topical and systematic massage.

Effect of Polygonati Sibirici Rhizoma on Cell Viability in Human Glioma Cells

  • Kim, Min-Soo;Jeong, Ji-Cheon
    • The Journal of Korean Medicine
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    • v.29 no.1
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    • pp.95-105
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    • 2008
  • Objectives : Although herbal medicines containing flavonoids have been reported to exert anti-tumor activities, it has not been explored whether Hwang-Jeong (Polygonati sibirici Rhizoma, PsR) exerts anti-tumor activity in human glioma. The present study was therefore undertaken to examine the effect of PsR on cell viability and to determine its underlying mechanism in A172 human glioma cells. Methods : Cell viability was estimated by MTT assay. Reactive oxygen species generation and mitochondrial membrane potential were measured by the fluorescence dyes. The phosphorylation of kinases was evaluated by western blot analysis and caspase activity was estimated using colorimetric assay kit. Results : PsR resulted in loss of cell viability in a dose- and time-dependent manner. PsR did not increase reactive oxygen species (ROS) generation and the PsR-induced cell death was also not affected by antioxidants, suggesting that ROS generation is not involved in loss of cell viability. Western blot analysis showed that PsR treatment caused rapid reduction in phosphorylation of extracellular signal-regulated kinase (ERK) without changes in p38 and Jun-NH2-terminal kinase (JNK). U0126, an inhibitor of ERK, increased the PsR-induced cell death, but inhibitors of p38 and JNK did not affect the cell death. PsR induced depolarization of mitochondrial membrane potential. Caspase activity was not stimulated by PsR and caspase inhibitors did not prevent the PsR-induced cell death. Conclusion : Taken together, these findings suggest that PsR results in human glioma cell death through caspaseindependent mechanisms involving down-regulation of ERK.

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Gold Nanoparticles Inhibit AGEs Induced Migration and Invasion in Bovine Retinal Endothelial Cells (소망막내피세포에서 금 나노입자의 최종당화산물에 의한 세포 이동 및 침윤성 억제 효과)

  • Chae, Soo-Chul
    • Korean Journal of Environmental Biology
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    • v.28 no.1
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    • pp.8-13
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    • 2010
  • This study aimed the role of gold nanoparticles (AuNP) in advanced glycation end-products (AGEs) induced migration and invasion in bovine retinal endothelial cells (BRECs). BRECs were isolated from the retina. Cell viability was confirmed by the MTT assay. In vitro wound migration assay was performed to investigate the migration of BRECs. In vitro tube formation was measured by on-gel system. Apoptosis induced by AuNP was confirmed by caspase-3 assay. AGE-bovine serum albumin (BSA) demonstrated increase of cell migration and proliferation in BRECs. In addition, AuNP regardless of the existence of AGE-BSA suppressed proliferation, migration, and angiogenesis. AuNP suppressed AGE-BSA induced migration and invasion, and induced apoptosis through caspase-3. As a results, AuNP have a potential anti-angiogenic effect for AGE-induced angiogenesis in vitro and offer possibility for the treatment of diabetic retinopathy.

Effect of Mitomycin-C on Rabbit Conjunctival Fibroblast Proliferation and Apoptosis (마이토마이신이 가토 결막섬유아세포의 증식 및 사멸에 미치는 효과)

  • Kwon Young-Sam;Jang Kwang-Ho
    • Journal of Veterinary Clinics
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    • v.23 no.2
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    • pp.153-157
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    • 2006
  • The purpose of this study was to determine the effects of mitomycin C on conjunctival fibroblast proliferation and apoptosis. Rabbit conjunctival fibroblast mono layers were treated with 48 hours application of mitomycin-C. The viability of cells was evauated by MTT assay. To examine the effect of mitomycin-C on the cell proliferation, immunocytochemistry for BrdU staning was performed. Then, we investigated whether mitomycin-C induced cell's apoptosis by TUNEL staining. As a result of MTT assay, the viability of cells was gradually inhibited in a dose dependent manner by mitomycin-C. The BrdU staining showed that mitomycin-C significantly suppressed the proliferation of conjunctival fibroblast at concentrations of 0.02% or more. When TUNEL assays were performed, the number of apoptotic cells increased 5.6-, 18.5-, and 33.8-fold compared with the control at 0.01, 0.02, and 0.04%, respectively, of mitomycin-C at 48 hours of exposure. Therefore, mitomycin-C may be useful as a regulator in the treatment of corneal diseases that manifest with scar formation and tumor of fibroblast expansion.