• Title/Summary/Keyword: MTT reduction assay

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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 on Pancreatic Beta Cells and Nerve Cells by Low LET X-ray (Low LET X-ray가 췌장 ${\beta}$ 세포와 신경세포에 미치는 효과)

  • Park, Kwang-Hun;Kim, Kgu-Hwan
    • Journal of radiological science and technology
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    • v.37 no.1
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    • pp.21-28
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    • 2014
  • Cultured pancreatic beta cells and nerve cells, it is given normal condition of 10% FBS (fetal bovine serum), 11.1 mM glucose and hyperglycemia codition of 1% FBS, 30 mM glucose. For low LET X-ray irradiated with 0.5 Gy/hr dose-rate(total dose: 0.5 to 5 Gy). Survival rates were measured by MTT assay. When non irradiated, differentiated in the pancreatic beta cells experiment is hyperglycemia conditions survival rate compared to normal conditions survival rate seemed a small reduction. However increasing the total dose of X-ray, the survival rate of normal conditions decreased slightly compared to the survival rate of hyperglycemia conditions, the synergistic effect was drastically reduced. When non irradiated, undifferentiated in the nerve cells experiment is hyperglycemia conditions survival rate compared to normal conditions survival rate seemed a large reduction. As the cumulative dose of X-ray normal conditions and hyperglycemia were all relatively rapid cell death. But the rate of decreased survivals by almost parallel to the reduction proceed and it didn't show synergistic effect.

Cytotoxic Effect of Zostera asiatica on Growth of Human Cancer Cells (왕거머리말의 인체 암세포에 대한 세포독성 효과)

  • Hong, Joo-Wan;Jung, Myung-Eun;Lee, Jung-Im;Kim, Ho-Jun;Chang, Jae-Soo;Seo, Young-Wan
    • KSBB Journal
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    • v.27 no.4
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    • pp.227-231
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    • 2012
  • Whole plants of Zostera asiatica were extracted twice with acetone/methylene chloride (A+M) and methanol (MeOH) in turn. The combined crude extracts were evaporated in vacuo and then the residue was partitioned between water and methylene chloride. The aqueous layer was fractionated into $H_2O$ and n-butanol and then the organic layer was also fractionated into 85% aq. MeOH and n-hexane, successively. The crude extracts and their solvent fractions were evaluated for their inhibitory effect on growth of human cancer cells AGS, HT-29, MCF-7, and HT-1080 cells by MTT reduction assay. Among samples tested, 85% aq. MeOH and n-hexane fractions showed strong cytotoxic effect against AGS, HT-29, and MCF-7 cells. On the other hand, for HT-1080 cell, 85% aq. MeOH fraction exhibited the strongest cytotoxic effect.

Cytotoxic Activity of the Inula japonica Extracts Against Several Human Cancer Cell Lines In Vitro (선복화 (Inula japonica)추출물의 암세포주에 대한 In Vitro 세포독성)

  • Cha, Mi-Ran;Kim, Ju-Young;Hwang, Ji-Hwan;Park, Hae-Ryong
    • Korean Journal of Pharmacognosy
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    • v.37 no.3
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    • pp.130-135
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    • 2006
  • The present study describes the preliminary evaluation of the cytotoxic activity of the extracts from Inula japonica. I. japonica was extracted with methanol, ethanol, acetone, and water, and then cytotoxic activity of these extracts were evaluated. The cytotoxic activity of each extract was assessed by the MTT-dye reduction assay. Both ethanol and acetone extracts from I. japonica showed the cytotoxic activity against the HT-29 human colon cancer cells. Furthermore, the ethanol extract was fractionated with n-hexane, diethyl ether, ethyl acetate, and water according to degree of Polarity, The diethyl ether fraction showed the highest cytotoxic activity against HT-29 cells, but the other fractions showed low cytotokic activity. In addition, diethyl ether layer also showed the cytotoxic activity against various tumor cells, such as human colon carcinoma SW620, human cervix adenocarcinoma HeLa, and human breast adenocarcinoma MCF-7 cells as well as HT-29 cells. These studies support that extracts of I. japonica may be a potential candidate as possible chemotherapeutic agent against human cancer.

Protective Effects of Seok-Jeong on the Toxicity of Cadmium in Neuronal Cells (뇌신경세포에서 토양미생물 발효추출액인 석정의 카드뮴 독성에 대한 방어효과)

  • 홍순해;안성희;장봉기;박종안;이종화
    • YAKHAK HOEJI
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    • v.47 no.2
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    • pp.85-92
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    • 2003
  • Seok-jeong (SJ) is a solution of various metal ions and numerous other organic substances produced through extraction and fermentation of herbs and soil using geo-microbes, and it has been shown to improve symptoms of senile dementia. In this study, we investigated the protective effects of SJ against neurotoxicity of cadmium in HT22 hippocampal neuron cell line. SJ significantly protected from the cadmium-induced decreased cell viability measured by MTT assay (p<0.01). The protective effects of SJ against cadmium toxicity were confirmed through observing morphological changes using inverted microscope. Additionally, SJ significantly repressed the formation of lipid peroxidation induced by high concentration of cadmium, and likewise, significantly repressed the reduction of glutathione by cadmium in HT22 cells. Vitamin C at the concentration found in SJ did not show any protective effect against cadmium toxicity in HT22 cells, indicating that vitamin C may not have a major role in the protective mechanism of SJ. Taken together, these results suggest that SJ may be a valuable agent for the protection of cadmium toxicity on the neuronal cells, and that the mechanism of the action of SJ may be due to reduced lipid peroxidation and increased glutathione level.

Anti-oxidative Effect of Some Plant Extracts Against Nitric Oxide-induced Oxidative Stress on Neuronal Cell (Nitric oxide에 의해 산화적 스트레스를 받은 Neuronal cell에 항산화 효과를 가지는 수종 생약추출물의 검색)

  • Koo, Uk;Lee, Hak-Ju;Lee, Dong-Ho;Lee, Hyun-Jung;Ham, Ah-Rom;Cho, Eun-Young;Mar, Woong-Chon
    • Korean Journal of Pharmacognosy
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    • v.39 no.4
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    • pp.290-294
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    • 2008
  • The objective of this study is screening the anti-oxidative effects of several plant MeOH extracts against oxidative stress in Neuroblastoma cell. Oxidative stress has been implicated in the pathogenesis of many neurotoxicity, neurodegenerative disorders and cell death. This oxidative stress is generated by ROS (Reactive Oxygen Species) such as nitric oxide, nitrogen dioxide, peroxyl, superoxide ($O_2^-$), hydroxyl, alkoxyl. So, in the present study, we induced oxidative stress by treatment of sodium nitroprusside (2.5 mM) in human neuroblastoma SH-SY5Y cell which was treated samples before 24hr, and cell viability was measured by MTT reduction assay. Of those tested, the extracts of Paeonia japonica (roots), Eucommia ulmoides (炒)(barks), Paeonia japonica (曝乾)(roots), Phyllostachys bambusoides (stems), Polygala tenuifolia (去心, 炒)(roots), Paeonia japonica (roots), Polygala tenuifolia (roots), Machilus thunbergii (barks), Mallotus japonicus (leaves), Poria cocos (whole), Sophora flavescens (roots), Angelica tenuissima (roots), Angelica gigas (當歸尾)(roots) showed anti-oxidative effects[$EC_{50}$<15.20 ${\mu}g$/ml(Carnosine:Positive control)]in dose dependent manner.

Antioxidant and Neuronal Cell Protective Effect of Purple Sweet Potato Extract (자색고구마 추출물의 항산화 효과 및 신경세포 보호효과)

  • Kwak, Ji-Hyun;Choi, Gwi-Nam;Park, Ju-Hee;Kim, Ji-Hye;Jeong, Hee-Rok;Jeong, Chang-Ho;Heo, Ho-Jin
    • Journal of agriculture & life science
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    • v.44 no.2
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    • pp.57-66
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    • 2010
  • The antioxidant and neuronal cell protective effects of water extract from purple sweet potato were investigated. The total phenolics and monomeric anthocyanin contents of purple sweet potato extract were 44.25 mg/g and 2,394 mg/L, respectively. The antioxidant activities of purple sweet potato extract were evaluated using various antioxidant tests, including 1,1-diphenyl- 2-picrylhydrazyl (DPPH), 2,2'-azino- bis-(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) radical scavenging activities, ferric reducing/antioxidant power (FRAP) and reducing power. In these assays, the extract of purple sweet potato presented significant radical scavenging activities, FRAP, and reducing power in a dose-dependent manner. MTT {3-(4,5-dimethylthiazol-2-yl)- 2,5-diphenyl- tetrazoliumbromide} reduction assay showed significantly increase in cell viability when PC12 cells were pretreated with purple sweet potato extract. Because oxidative stress is also known to increase neuronal cell membrane breakdown, we further investigated by lactate dehydrogenase (LDH) and neutral red uptake assay. Purple sweet potato extract inhibited oxidative stress-induced membrane damage in neuronal cells. Therefore, these data results demonstrated that water extract of purple sweet potato have antioxidant activity and neuronal cell protective effect thus it has great potential as a natural source for human health.

Cytotoxic and Apoptotic Activities of Tussilago farfara Extract in HT-29 Human Colon Cancer Cells

  • Lee, Mi-Ra;Cha, Mi-Ran;Jo, Kyung-Jin;Yoon, Mi-Young;Park, Hae-Ryong
    • Food Science and Biotechnology
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    • v.17 no.2
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    • pp.308-312
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    • 2008
  • The flower buds of Tussilago farfara (TF) have been traditionally used in oriental medicine for the treatment of bronchitis and asthma. In our study, the primary objective was to determine the mechanisms that are inherent to TF-induced cytotoxicity and apoptosis, using the methanolic extract of TF (TFM) in HT-29 human colon cancer cells. We found that TFM-induced induced cytotoxicity in HT-29 cells in a dose-dependent manner. This effect was verified via an MTT reduction assay, an lactate dehydrogenase (LDH) release assay, and a colony formation assay. Interestingly, we also detected apoptotic bodies on Hoechst staining, and attempted to determine whether TFM-induced apoptosis involved the caspase pathway using a caspase-3/7 activity assay. Overall, the results indicate that TFM contain chemotherapeutic agents and potential candidates use for against human colon cancer cells.

Apoptosis Induction in Human Leukemic Promyelocytic HL-60 and Monocytic U937 Cell Lines by Goniothalamin

  • Petsophonsakul, Ploingarm;Pompimon, Wilart;Banjerdpongchai, Ratana
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.5
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    • pp.2885-2889
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    • 2013
  • Goniothalamin is an active compound extracted from Goniothalamus griffithii, a local plant found in northern Thailand. Goniothalamin inhibits cancer cell growth but is also toxic to normal cells. The aims of this study were to identify the cytotoxic effect of goniothalamin and the mechanism of cell death in human HL-60 and U937 cells. Cytotoxicity was determined by MTT assay and cell cycle profiles were demonstrated by staining with propidium iodide (PI) and flow cytometry. Apoptosis was confirmed by staining with annexin V-FITC/propidium iodide (PI) and flow cytometry. Reduction of mitochondrial transmembrane potential was determined by staining with dihexyloxacarbocyanine iodide and flow cytometry and expression of Smac, caspase-8 and -9 was demonstrated by Western blotting. Goniothalamin inhibited growth of HL-60 and U937 cell lines. An increase of SubG1 phase was found in their cell cycle profiles, indicating apoptosis as the mode of cell death. Apoptosis was confirmed by the flip-flop of phosphatidylserine using annexin V-FITC/PI assay in HL60 and U937 cells in a dose response manner. Furthermore, reduction of mitochondrial transmembrane potential was found in both cell types while expression of caspase-8, -9 and Smac/Diablo was increased in HL-60 cells. Taken together, our results indicate that goniothalamin-treated human leukemic cells undergo apoptosis via intrinsic and extrinsic pathways.

Effect of Aceton Extract from Styela Clava on Oxidative DNA Damage and Anticancer Activity (미더덕 아세톤 추출물이 산화적 DNA 손상억제 및 암세포 독성에 미치는 영향)

  • Seo, Bo-Young;Jung, Eun-Sil;Kim, Ju-Young;Park, Hae-Ryong;Lee, Seung-Cheol;Park, Eun-Ju
    • Applied Biological Chemistry
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    • v.49 no.3
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    • pp.227-232
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    • 2006
  • Styela clava (also called as rough sea squirt or leathery tunicate) is regarded as native to the northwest Pacific region including Korea and widely distributed in parts of northwestern Europe, North America and Australia. To evaluate Styela clava as a potential bioactive agent, the antioxidant activity of aceton extracts from Styela clava (whole, substance and tunic) was tested by measuring inhibitory effect of $H_2O_2$ induced DNA damage using comet assay. Also, anticancer activity on human colon cancer cell (HT-29) was investigated by MTT reduction assay. The $200\;{\mu}M$ $H_2O_2$ induced DNA damage was inhibited with Styela clava aceton extract in dose dependent manner in human leukocytes. The maximum inhibition was by 62.8, 62.1 and 78.3% at the concentration of $50\;{\mu}g/ml$ of whole, substance and tunic extracts, respectively. The aceton extracts from S. clava were also found to inhibit the growth of human colon cancer cell. The cell proliferation rates decreased to 26.9, 30.6 and 12.0% at the concentration of $500\;{\mu}g/ml$ of whole, substance and tunic extracts, respectively. These results support that aceton extracts from S. clava may be a potential candidate as a possible antimutagenic and chemotherapeutic agent.