• Title/Summary/Keyword: MTT assay

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AN EXPERIMENTAL STUDY ON THE CHANGE OF THE RADIOSENSITIVITY OF SEVERAL TUMOR CELL LINES AND PRIMARY CULTURED GINGIVAL FIBROBLAST (수종의 암세포주와 치은섬유아세포에서 방사선의 양과 분할조사에 따른 세포활성도와 독성의 변화에 관한 연구)

  • Lee Sam-Sun;You Dong-Soo
    • Journal of Korean Academy of Oral and Maxillofacial Radiology
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    • v.27 no.1
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    • pp.107-122
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    • 1997
  • Radiation sensitivity data was generated for two human cancer cell lines(KB, RPMI 2650) and human primary gingival fibroblast was tested three times using a viable cell number counting with a hemocytometer, MTT(3-[4,5-Dimethylthiazol2-yl]-2,5-diphenyl tetrazolium bromide) assay, and LDH(Lactate dehydrogenase) assay. Single irradiation of 2, 4, 6, 10, 15, 20Gy were applied to the tumor cell lines and the primary cultured gingival fibroblast The two fractions of 4Gy and 10Gy were seperated with a 4 hour time interval. The irradiation was done with 241.5cGy/min dose rate using /sup 137/Cs MK cell irradiator at room temperature. The obtained results were as followed : 1. There was significantly different viable cell numbers as the amount of radiation dose on the tested cells were cell number counted with a hemocytometer. In fractions, there were more viable cells remaining. 2. Phase-contrast microscopically, radiation-induced morphologic changes were pronounced on the tumor cells, however, almost no differences on the gingival fibroblast. 3. There was significantly different absorbance at 2Gy on RPMI 2600, 4Gy on KB and GF in MTT assay. In fractions, the absorbance was significantly higher on KB. 4. The level of extracellular LDH activity in the experimental group was significantly higher in the 2-4Gy than the control group. 5. The total level of extracellular and intracellular LDH activity was decreased as increased amounts of radiation dose was applied.

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Cytotoxic Effect of Inonotus obliquus Composition in HCT-15 Human Colon Cancer Cells and AGS Gastric Cancer Cells (대장암 세포암종 HCT-15 세포 및 위암 세포암종 AGS 세포에서 차가버섯 조성물에 의한 세포생육 억제 효과)

  • 차재영;전병삼;문재철;유지현;조영수
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.33 no.4
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    • pp.633-640
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    • 2004
  • This study was performed to investigate the cytotoxic effect of the water-extract from Chaga mushroom (Inonotus obliquus) compositions containing powdered green tea in HCT-15 human colon carcinoma, AGS human gastric carcinoma and NIH3T3 mouse normal fibroblast cells using viable cell count and MTT assay. The water-extract from Chaga mushroom compositions induced inhibitory effects on proliferation of HCT-15 and AGS cells in the MTT assay and viable cell count. However, mouse normal NIH3T3 cells were exhibited 80% survival under the same condition. Chaga mushroom compositions showed highly antiproliferative effect in human cancer cell line HCT-15 and AGS, but not in mouse normal cell line NIH3T3. These results suggest that Chaga mushroom (Inonotus obliquus) compositions containing powdered green tea are the candidate for chemoprevention in colon and gastric cancer.

Study of Antioxidant Effects of Platycladi Semen on Male Reproductive cells (백자인(柏子仁)이 수컷 생쥐의 생식세포에 미치는 항산화 효과 연구)

  • Kim, Ju-Sung;Hong, Min-Jung;Kim, Do-Rim;Park, Eun-Hwa;Chang, Mun-Seog;Park, Seong-Kyu
    • The Korea Journal of Herbology
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    • v.27 no.2
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    • pp.37-42
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    • 2012
  • Objectives : The purpose of this study is to examine the antioxidant effects on male mouse reproductive cells of the extract of Platycladi Semen. Methods : The extract was studied for diphenyl-picryl-hydrazyl (DPPH) radical scavenging activity, cell viability by a modified MTT assay, the effects on $H_2O_2$-induced cytotoxicity by MTT assay, lipid peroxidation by malondialdehyde (MDA) formation and super oxide dismutase (SOD), respectively. Results : The results showed that the extract scavenged DPPH radical in a dose-dependent manner by up to 74.87%. The cell viability of the extract was within 72~96% on Leydig cells and GC-2 cells at concentrations of 1, 5, 10, 50 and 100 ug/ml. The hydrogen peroxide-induced cytotoxicity of Leydig cells was protected to 72.09% by the extract at concentration of 100 ${\mu}g/ml$. The hydrogen peroxide-induced lipid peroxidation of MDA formation was decreased to 1.80 and 1.65 nmoles/mg protein by the extract at concentrations of 50 and 100 ${\mu}g/ml$. The extract at all concentrations, SOD activity was not significantly changed. Conclusions : In conclusion, the extract of Platycladi Semen has antioxidant effects on Leydig cells and protect male reproductive system against oxidative stress.

A Study on the Cytotoxicity of Lead in Cultured Fibroblasts (납(Pb)이 배양 섬유모세포에 미치는 세포독성에 관한 연구)

  • 정연태;최민규;김정중;문연자;김재민;백순기
    • Environmental Mutagens and Carcinogens
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    • v.15 no.2
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    • pp.122-130
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    • 1995
  • In order to evaluate the cytotoxicity of lead in cultures of Balb/c mouse 3T3 cell line, various cytotoxic assays were carried out after expose cells to various concentrations of lead nitrate. Cytotoxic assays using this study were included NR assay, MTT assay, measurement of LDH and protein, synthetic rate of DNA and UDS. Intrace!!ular Ca$^{2+}$ level was also measured. Light and electron microscopic studies were done for morphological changes of lead-treated cell cultures. The results were as follows; 1. The absorbances of NR and MTT were decreased dose-dependently, and NR, and MTT, values of lead nitrate were 3.4 mM and 1.5 mM, respectively. 2. Amount of LDH released into the medium was increased in dose-dependently and LDH activity at 5 mM concentration of lead nitrate was increased to 335 % of control. 3. Amount of total protein was decreased dose-dependently, and which was half of control at 2 mM concentration of lead nitrate. 4. The synthetic rate of DNA was decreased dose-dependently, and also which was remarkably decreased at 3 mM and 5 mM concentrations of lead nitrate. 5. The synthetic rate of UDS was increased at 1 mM concentration of lead nitrate, but which was remarkably decreased at 3 mM and 5 mM concentrations of lead nitrate. 6. Intrace!lular Ca$^{2+}$ level was remarkably increased at 1 mM concentration of lead nitrate, compared with control. 7. In light microscopy, number of cells and processes were decreased according to the increase of dosage of lead nitrate. Electron microscopic findings showed that many vacuoles and cisternal dilatation of rough endoplasmic reticulum were seen in the cytoplasm at 1 mM concentration of lead nittale. From the above results, high dosage treatment of lead nitrate (>3 mM) damaged genetic malerials and it also showed cytotoxicity in mouse 3T3 cell line cultures by injury of cell organelles and Ca$^{2+}$ channel.

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Extracts of Centaurea bornmuelleri and Centaurea huber-morathii inhibit the growth of colon cancer cells in vitro

  • Sarker, Satyajit Dey;Shoeb, Mohammad;Celik, Sezgin;Jaspars, Marcel;Nahar, Lutfun;Kong-Thoo-Lin, Paul;MacManus, Stephen M
    • Advances in Traditional Medicine
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    • v.7 no.4
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    • pp.336-340
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    • 2007
  • Plants from the genus Centaurea (C.) (Family: Asteraceae alt. Compositae), widely distributed in Asia, Europe and North America, have traditionally been used in the treatment of various ailments. As a part of our on-going studies on the plants from the genus C. for their phytochemistry and biological activities, extracts of the seeds of Turkish endemic C. species, C. bornmuelleri and C. huber-morathii, were tested for their cytotoxicity towards the CaCo2 colon cancer cell line as well as for the toxicity towards the brine shrimps, using the MTT and the brine shrimp lethality assays, respectively. Among the extracts, the MeOH extract of these plants showed significant toxicity towards the brine shrimps ($LD_{50}=55.2{\times}10^{-2}\;and\;42.4{\times}10^{-2}mg/ml$, respectively). The MeOH extract of both C. species also inhibited the growth of CaCo2 colon cancer cells in the MTT assay ($IC_{50}$=29.9 and 33.0 g/ml, respectively). As the most prominent activities in both assays were observed with the MeOH extracts, it can be assumed that the compound(s) responsible for these activities are polar in nature.

Protective Effects of Cornu Saigae Tataricae Extracts on Cultured Spinal Motor Neurons Damaged by Oxygen Free Radical (산소자유기에 의한 척수운동세포 독성에 대한 영양각 추출물의 방어효과)

  • Kang Gil Seong;Kwon Kang Beom;Ryu Do Gon
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.17 no.5
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    • pp.1202-1207
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    • 2003
  • In order to clarify the neuroprotective effect of Cornu Saigae Tataricae(CST) water extract on cultured mouse spinal motor neuron damaged by hydrogen peroxide (H₂O₂), MTT [3-(4,5-dimethylthiazole-2-yl)- 2,5-diphenyltetrazolium bromide] assay, LDH (Lactate Dehydrogenase) activity assay and SRB (Sulforhodamine B) assay were carried out after the cultured mouse spinal motor neuron were preincubated with various concentrations of CST water extract for 3 hours prior to exposure of hydrogen peroxide Cell viability of cultured mouse spinal motor neurons exposed to various concentrations of hydrogen peroxide for 6 hours was decreased in a dose-dependent manner. MTT50 values were 40 uM hydrogen peroxide. Cultured mouse spinal motor neurons in the medium containing various concentration of hydrogen peroxide for 6 hours showed increasing of LDH activity and decreasing of total protein synthesis. We know that hydrogen peroxide was toxic on cultured spinal motor neurons. Pretreatment of CST water extract for 3 hours following hydrogen peroxide prevented the hydrogen peroxide-induced neurotoxicity such as increasing of LDH activity and decreasing of total protein synthesis. These results suggest that hydrogen peroxide shows toxic effect on cultured spinal motor neurons and CST water extract is highly effective in protecting the neurotoxicity induced by hydrogen peroxide.

Comparative Antitumor Activity of Water Extracts from Fruiting Body of Phellinus linteus, Phellinus baumii and Phellinus gilvus (Phellinus linteus, Phellinus baumii 및 Phellinus gilvus 자실체 추출물의 항암효과 비교)

  • 배재성;황미현;장광호;이만휘;이근우;조우식;최성국;윤효인;임종환
    • Toxicological Research
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    • v.20 no.1
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    • pp.37-42
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    • 2004
  • This study was undertaken to investigate comparative anti-tumor activity of water extracts of Phellinus gilvus (PGE), Phellinus linteus (PLE), and Phellinus baumii (PBE) in vitro. The anti-tumor activity in the present study was evaluated by sulforhodamine B (SRB) and microtetrazolium (MTT) assay in terms of cell survival level. The tumor cells (sarcoma 180 and P388) were treated with PGE, PLE, and PBE (7.5, 15, and 30 $\mu\textrm{g}$/ml) and Doxorubicin (DOX) (0.001~10 $\mu\textrm{M}$). The results showed that DOX, PGE, and PLE inhibited proliferation showing a dose-dependent manner against both tumor cells. However, PBE was inhibited by the only 30 $\mu\textrm{g}$/ml in both cells proliferation. In conclusion, all of PGE, PLE, and PBE used in this study have shown anti-tumor activity against both sarcoma 180 and P388. Among them, PLE was the most effective in anti-tumor activity against sarcoma 180 (p<0.05) and PGE was against P388 in SRB assay. PLE, however, was against P388 (p<0.05) in MTT assay.

Antioxidant Effects of Corni Fructus in GC-1 Cells (산수유(山茱萸)가 남성 생식세포 GC-1의 항산화에 미치는 영향)

  • Oh Myung-Sook;Kim Do-Rim;Sung Eun-Jin;Chang Mun-Seog;Park Seong-Kyu
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.19 no.6
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    • pp.1541-1545
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    • 2005
  • The purpose of this study is to examine the antioxidant activity in the germ cells of the extract of Corm fructus. The extract was studied for diphenyl-picryl-hydrazyl (DPPH) radical scavenging activity, GC-1 cell viability by a modified MTT assay, the effects on $H_2O_2$-induced cytotoxicity by MTT assay and lipid perixidation by malondialdehyde (MDA) formation, respectively. The results showed that the extract scavenged DPPH radical with the IC50 being $200{\mu}g/mL$. The extract at concentrations of $10-500{\mu}g/ml$ showed dose-dependent in growth of GC-1 cell. $H_2O_2$-induced cytotoxicity (63.0%) was blocked by the extract (10, 50, 100, 250 and $500{\mu}g/ml$) concentration-dependently. Furthermore, the extract (50, 100 and $250{\mu}g/ml$) also displayed a dose-dependent reduction of MDA formation on $H_2O_2$-induced lipid peroxidation. In conclusion, the extract of Corm fructus has potent antioxidant activity.

DNA microarray analysis of gene expression of MC3T3-E1 osteoblast cell cultured on anodized- or machined titanium surface

  • Park, Ju-Mi;Jeon, Hye-Ran;Pang, Eun-Kyoung;Kim, Myung-Rae;Kang, Na-Ra
    • Journal of Periodontal and Implant Science
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    • v.38 no.sup2
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    • pp.299-308
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    • 2008
  • Purpose: The aim of this study was to evaluate adhesion and gene expression of the MC3T3-E1 cells cultured on machined titanium surface (MS) and anodized titanium surface (AS) using MTT test, Scanning electron micrograph and cDNA microarray. Materials and Methods: The MTT test assay was used for examining the proliferation of MC3T3-E1 cells, osteoblast like cells from Rat calvaria, on MS and AS for 24 hours and 48 hours. Cell cultures were incubated for 24 hours to evaluate the influence of the substrate geometry on both surfaces using a Scanning Electron Micrograph (SEM). The cDNA microarray Agilent Rat 22K chip was used to monitor expressions of genes. Results: After 24 hours of adhesion, the cell density on AS was higher than MS (p < 0.05). After 48 hours the cell density on both titanium surfaces were similar (p > 0.05). AS had the irregular, rough and porous surface texture. After 48 hours incubation of the MC3T3-E1 cells, connective tissue growth factor (CTGF) was up-regulated on AS than MS (more than 2 fold) and the insulin-like growth factor 1 receptor was down-regulated (more than 2 fold) on AS than MS. Conclusion: Microarray assay at 48 hours after culturing the cells on both surfaces revealed that osteoinductive molecules appeared more prominent on AS, whereas the adhesion molecules on the biomaterial were higher on MS than AS, which will affect the phenotype of the plated cells depending on the surface morphology.

Effect of Epimedium Koreanum Nakai on GO-Induced Neurotoxicity in Cultured Mouse Spinal Dorsal Root Ganglion Neurons (Glucose Oxidase에 의(依)하여 손상(損傷)된 배양척수감각신경절세포(培養脊髓感覺神經節細胞)에 대(對)한 음양곽(淫羊藿)의 효과(效果))

  • Park Seung-Taeck;Lee Ho-Sub;Yun Yong-Gap;Park Byung-Rim
    • Herbal Formula Science
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    • v.7 no.1
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    • pp.143-151
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    • 1999
  • To evaluate the neurotoxic effect of oxygen radicals in cultured mouse spinal dorsal root ganglion(DRG) neurons, cytotoxicity was determined by MTT assay after cultured DRG neurons were grown in the media containing various concentrations of glucose oxidase(GO). In addition, neuroprotective effect of herb extract, Epimedium Koreanum Nakai was examined by MTT assay in cultured DRG neurons. Cell viability of cultured DRG neurons was remarkably decreased by GO in dose- and time-dependent manner, and Epimedium Koreanum Nakai protected remarkably GO-induced neurotoxicity in these cultures. From the above results, it is suggested that oxygen radicals is toxic in cultured mouse DRG neurons, and herb extracts such as Epimedium Koreanum Nakai are effective in prevention of the neurotoxicity induced by oxygen radicals in cultured mouse DRG neurons.

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