• Title/Summary/Keyword: cell viability assay

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Cytotoxicity of Nigella Sativa Seed Oil and Extract Against Human Lung Cancer Cell Line

  • Al-Sheddi, Ebtesam Saad;Farshori, Nida Nayyar;Al-Oqail, Mai Mohammad;Musarrat, Javed;Al-Khedhairy, Abdulaziz Ali;Siddiqui, Maqsood Ahmed
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.2
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    • pp.983-987
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    • 2014
  • Nigella sativa (N sativa), commonly known as black seed, has been used in traditional medicine to treat many diseases. The antioxidant, anti-inflammatory, and antibacterial activities of N sativa extracts are well known. Therefore, the present study was designed to investigate the anticancer activity of seed extract (NSE) and seed oil (NSO) of N sativa against a human lung cancer cell line. Cells were exposed to 0.01 to 1 mg/ml of NSE and NSO for 24 h, then percent cell viability was assessed by 3-(4, 5-dimethylthiazol-2yl)-2, 5-biphenyl tetrazolium bromide (MTT) and neutral red uptake (NRU) assays, and cellular morphology by phase contrast inverted microscopy. The results showed NSE and NSO significantly reduce the cell viability and alter the cellular morphology of A-549 cells in a concentration dependent manner. The percent cell viability was recorded as 75%, 50%, and 26% at 0.25, 0.5, and 1 mg/ml of NSE by MTT assay and 73%, 48%, and 23% at 0.25, 0.5, and 1 mg/ml of NSE by NRU assay. Exposure to NSO concentrations of 0.1 mg/ml and above for 24 h was also found to be cytotoxic. The decrease in cell viability at 0.1, 0.25, 0.5, and 1 mg/ml of NSO was recorded to be 89%, 52%, 41%, and 13% by MTT assay and 85%, 52%, 38%, and 11% by NRU assay, respectively. A-549 cells exposed to 0.25, 0.5 and 1 mg/ml of NSE and NSO lost their typical morphology and appeared smaller in size. The data revealed that the treatment of seed extract (NSE) and seed oil (NSO) of Nigella sativa significantly reduce viability of human lung cancer cells.

Cytotoxic Effect of Hexavalent Chromium on C6 Glioma Cells

  • Suk Seung-Han
    • Biomedical Science Letters
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    • v.12 no.3
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    • pp.261-265
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    • 2006
  • Toxic effect of Hexavalent chromium $(CrO_3)$ on various cells and organs has been well recognized. However, the mechanism and degree of cytotoxicity of $CrO_3$ remain unclear. This study was performed to examine the cytotoxicity of $CrO_3$ on $C_6$ glioma cells by measuring cell viability. The XTT assay, one of the sensitive methods to determine the cell viability, was taken to examine the viability of glioma cells treated with $CrO_3$. In this study, not only decreased the number of glioma cells but morphologic changes of them were noted and cell viability decreased in a time and dose-dependent manner after treated with various concentrations of $CrO_3$ for 48hours. $IC_{90}\;and\;IC_{50}$ values in XTT assay were determined at $25{\mu}M\;and\;55{\mu}M$ $CrO_3$, respectively. These results suggest that Hexavalent chromium has a highly cytotoxic effect and has a time and dose-dependent cytotoxicity on $C_6$ glioma cells.

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EFFECT OF CULTURE SUPERNATANT OF BACTERIA ISOLATED FROM INFECTED ROOT CANALS ON CELL LINES (감염 근관에서 분리한 세균 배양액이 배양된 세포에 미치는 영향)

  • Im, Mi-Kyung
    • Restorative Dentistry and Endodontics
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    • v.22 no.2
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    • pp.761-768
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    • 1997
  • Microorganisms and their by-products are considered to be the major causes of pulpal and periapical pathosis. The role of microorganisms in endodotic infection has been studied regarding the prevalence of particular organisms found in root canal and periapical lesions. The aim of this study was to investigate the effect of culture supernatants of several oral microorganims isolated from infected root canals on the viability of cultured cell lines using colorimetirc assay. S. simulans, S. sciuri, E. faecium, S. intermedius, S. mitis, S. sanguis and S. uberis were incubated in Todd-Hewitt broth for 16 hours. 20 and 100ul of filtered bacterial cell culture supernatants were added to MK and Hep-2 cells. Cell viability was measured using MIT colorimetric assay. 20ul and 100u1 of S. sanguis supernatant showed significant cytotoxicity compared to control on MK cells. 100ul of S. sanguis supernatant significantly depressed viability of HEp-2 cells. E. faecium and S. intermedius did not affect the viability of MK and HEp-2 cells.

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Effect of Lycii cortex radicis Extraction on Glioma Cell Viability

  • Kim, Seang-Jae;Jeong, Ji-Cheon
    • The Journal of Korean Medicine
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    • v.30 no.6
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    • pp.17-26
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    • 2009
  • Objectives: Little information is available regarding the effect of Lycii cortex radicis (LCR) on cell viability in glioma cells. This study was therefore undertaken to examine the effect of LCR on cell survival in U87MG human glioma cells. Methods: Cell viability and cell death were estimated by MTT assay and trypan blue exclusion assay, respectively. Reactive oxygen species (ROS) generation was measured using the fluorescence probe DCFH-DA. Activation of Akt and extracellular signal-regulated kinase (ERK) and activation of caspase-3 were estimated by Western blot analysis. Results: LCR resulted in apoptotic cell death in a dose- and time-dependent manner. LCR increased reactive oxygen species (ROS) generation and LCR-induced cell death was also prevented by antioxidants, suggesting that ROS generation played a critical role in LCR-induced cell death. Western blot analysis showed that LCR treatment caused down-regulation of Akt and ERK. The LCR-induced cell death was increased by the inhibitors of Akt and ERK. Activation of caspase-3 was stimulated by LCR and caspase inhibitors prevented the LCR-induced cell death. Conclusion: These findings suggest that LCR results in human glioma cell death through a mechanism involving ROS generation, down-regulation of Akt and ERK, and caspase activation.

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Effect of Injin Butanol Fraction with Thin Layer Chromatography on Fas-mediated Apoptosis (인진butanol 분획의 TLC추출성분이 Fas-mediated Apoptosis에 미치는 영향)

  • 박용진;김영철;이장훈;우흥정
    • The Journal of Korean Medicine
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    • v.23 no.2
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    • pp.57-69
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    • 2002
  • Objective: The aim of this study is to investigate the effect of Injin butanol fractions with Thin Layer Chromatography on Fas-mediated Apoptosis. Method: Injin-butanol fraction separated by TLC. MIT assay, cell cycle analysis, Caspase-3 protease assay, DNA fragmentation assay and quantitative RT-PCR were performed to evaluate the effects of TLC extraction of lnjin-butanol fraction on cell viability, cell cycle progression and apoptosis. Results: Scopoletin, luteolin, apigenin and unknown powder was isolated by TLC. Fas-mediated apoptosis analysis shows that scopoletin has inhibiting function on apoptosis. Caspase- 3 protease assay analysis shows that scopoletin inhibits activity of caspase-3. Quantitative RT-PCR analysis shows that no activity on caspase-3, but apoptosis inhibition cytokine -Bcl-2- is activated, and apoptosis activating cytokine -Bax- is unactivated. Conclusion: These results show that each fraction of Injin-butanol TLC extraction, especially scopoletin, acts as a protective function on liver cell viability, and inhibitory function on apoptosis. (J Korean Oriental Moo 2002;23(2):57-69)

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Effects of Taeumin Chungsimyeunjatang on the Cerebral neurons injured by Hydrogen Peroxide (태음인(太陰人) 청심연자탕(淸心蓮子湯)이 Hydrogen Peroxide에 손상(損傷)된 백서(白鼠)의 대뇌신경세포(大腦神經細胞)에 미치는 영향(影響))

  • Ok, Yun-young;Ryu, Do-gon;Kim, Kyung-yo
    • Journal of Sasang Constitutional Medicine
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    • v.11 no.2
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    • pp.251-266
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    • 1999
  • 1. Purpose : The purpose of this study was to determine the effects of Chungsimyeunjatang on the cerebral neurons injured by hydrogen peroxide($H_2O_2$). 2. Methods : I observed cell viability in mouse cerebral neurons exposed to hydrogen peroxide by NR assay and MTT assay and determined lipid peroxidation and amounts of LDH release in mouse cerebral neurons exposed to hydrogen peroxide. After administration of Chungsimyeunjatang water extracts, I observed significant changes of cell viability, lipid peroxidation and amounts of LDH release in mouse cerebral neurons exposed to hydrogen peroxide. 3. Results : Hydrogen peroxide showed neurotoxicity. Cell viability in mouse cerebral neurons exposed to hydrogen peroxide decreased in NR assay and MTT assay. Lipid peroxidation and amounts of LDH release in mouse cerebral neurons exposed to hydrogen peroxide increased. Chungsimyeunjatang was very effective in blocking hydrogen peroxide-induced neurotoxicity.

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AN EXPERIMENTAL STUDY ON FAT CELL VIABLITY ACCORDING TO DIFFERENT HARVESTING TECHNIQUES (지방 채취 방법에 따른 지방 세포의 생존성에 대한 연구)

  • Lee, Won-Deok;Choi, Jin-Young
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.30 no.1
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    • pp.22-29
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    • 2008
  • Purpose: The purpose of this study is to test the efficacy of various methods of fat harvesting in animal model by viability comparison with assay including cell counting, MTT assay, and histologic evaluation. Materials and methods: New Zealand white rabbits experiments were used. Groin fat pads were subjected to different harvest method varying ingredients of solution(Experiment 1: T1 solution= lidocaine 1000mg/L, epinephrine 1mg/L, sodium bicarbonate 10mgEq/L, Triamcinolone 10mgEq/L; T2 solution=lidocaine 1000mg/L, epinephrine 1mg/L, sodium bicarbonate 0mgEq/L, Triamcinolone 0mgEq/L) and pressure exerted on harvesting with Luer-Lock syringe connected to suction cannula.(Experiment 2: P1 group=3cc intermittent pressure; P2 group=10cc sustained pressure) Fat cell viability was assessed with cell counting with a hemocytometer, MTT assay, and histologic evaluation. Results: Experiment 1 Cell count: T1=2.4/3.4/4.2, T2=9.6/8.4/7.2($\times10^5$ per mL); MTT assay: T1=0.516/0.41/0.453/0.412/0.421, T2=0.925/0.765/0.54/0.634/0.614 in 21 days(absorbance); Histology: T1 showed elongated and, different in size and shape, and ruptured adipocytes with only a few normal adipocytes whereas T2 showed central core of fat with almost intact fat cells Experiment 2 Cell count: P1=1.2/3.2/4.2, P2=1.2/2.4/3.8($\times10^5$ per mL); MTT assay:P1=0.256/0.245/0.258/0.21/0.264, P2=0.12/0.231/0.245/0.313/0.281 in 21 days(absorbance); Histology: P1 showed somewhat evenly distributed normal-looking fat cells and P2 showed relatively irregular shape of fat cells with small blood vessel amongst adiopocytes. Conclusion: Viability was higher in ‘modified tumescent solution’without sodium bicarbonate and triamcinolone and we also found no significantly different viability between using intermittent pressure and using sustained pressure. But in terms of initial viability of fat cell, we can assume that lower intermittent pressure would make better clinical results.

Portulaca oleracea Seed Oil Exerts Cytotoxic Effects on Human Liver Cancer (HepG2) and Human Lung Cancer (A-549) Cell Lines

  • Al-Sheddi, Ebtesam Saad;Farshori, Nida Nayyar;Al-Oqail, Mai Mohammad;Musarrat, Javed;Al-Khedhairy, Abdulaziz Ali;Siddiqui, Maqsood Ahmed
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.8
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    • pp.3383-3387
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    • 2015
  • Portulaca oleracea (Family: Portulacaceae), is well known for its anti-inflammatory, antioxidative, anti-bacterial, and anti-tumor activities. However, cytotoxic effects of seed oil of Portulaca oleracea against human liver cancer (HepG2) and human lung cancer (A-549) cell lines have not been studied previously. Therefore, the present study was designed to investigate the cytotoxic effects of Portulaca oleracea seed oil on HepG2 and A-549 cell lines. Both cell lines were exposed to various concentrations of Portulaca oleracea seed oil for 24h. After the exposure, percentage cell viability was studied by (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) (MTT), neutral red uptake (NRU) assays, and cellular morphology by phase contrast inverted microscopy. The results showed a concentration-dependent significant reduction in the percentage cell viability and an alteration in the cellular morphology of HepG2 and A-549 cells. The percentage cell viability was recorded as 73%, 63%, and 54% by MTT assay and 76%, 61%, and 50% by NRU assay at 250, 500, and $1000{\mu}g/ml$, respectively in HepG2 cells. Percentage cell viability was recorded as 82%, 72%, and 64% by MTT assay and 83%, 68%, and 56% by NRU assay at 250, 500, and $1000{\mu}g/ml$, respectively in A-549 cells. The 100 $100{\mu}g/ml$ and lower concentrations were found to be non cytotoxic to A-549 cells, whereas decrease of 14% and 12% were recorded by MTT and NRU assay, respectively in HepG2 cells. Both HepG2 and A-549 cell lines exposed to 250, 500, and $1000{\mu}g/ml$ of Portulaca oleracea seed oil lost their normal morphology, cell adhesion capacity, become rounded, and appeared smaller in size. The data from this study showed that exposure to seed oil of Portulaca oleracea resulted in significant cytotoxicity and inhibition of growth of the human liver cancer (HepG2) and human lung cancer (A-549) cell lines.

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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Effect of Allopurinol on Methylmercuric Chloride-Induced Cytotoxicity in $C_6$ Cultured Glioma Cells

  • Oh, Yong-Leol;Son, Byoung-Kwan
    • Biomedical Science Letters
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    • v.12 no.4
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    • pp.451-455
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    • 2006
  • It is demonstrated that inorganic mercury has cytotoxic effect on glial cells. Recently, oxygen radicals is involved in methylmercuric chloride (MMC)-induced cytotoxicity. But, the toxic mechanism of MMC is left unknown. The purpose of this study was to examine the cytotoxicity of MMC on $C_6$ glioma cells. The cytotoxicy was measured by cell viability using XTT assay in $C_6$ glioma cells. Colorimetric assay is regarded as a very sensitive screening method for the determination of the cell viability on various agents. In this study, MMC decreased cell viability according to the dose- and time dependent manners after $C_6$ glioma cells were grown with various concentrations of MMC for 48 hours. In the protective effect of allopurinol on MMC-induced cytotoxicity, allopurinol was effective in the prevention of MMC-induced cytotoxicity in these cultures. These results suggest that MMC has highly cytotoxic effect on $C_6$ glioma cells by the decrease of cell viavility, and free radical scavenger such as allopurinol was effective on organic mercury-induced cytotoxicity in these cultures.

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