• Title/Summary/Keyword: Sulforhodamine B (SRB) protein assay

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Cytotoxic Effect of Korean Traditional Prescriptions on the Human Gastric Cancer Cell Lines (한약처방제의 인체 위암 세포주에 대한 세포독성 효과에 관한 연구)

  • Kim, Eun-Hae;Eun, Young-Ah;Kang, Bong-Joo;Sung, Hyun-Jae;Park, Kap-Joo
    • Korean Journal of Pharmacognosy
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    • v.28 no.4
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    • pp.233-238
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    • 1997
  • ln order to search for antigastric cancer agents from Korean traditional prescriptions. We selected 41 traditional prescriptions, based on a review of the Korean traditional medicine books. Both boiling water and methanol extracts were tested, by means of the Sulforhodamine B (SRB) protein assay. Six of the 41 water extracts; #3, #34, #35, #38, #40, #41 showed efficacy against gastric cancer cell (AGS: Human gastric carcinoma, ATCC HTB 103). #3 inhibited 50% cancer cell growth1 at the concentration of $152\;{\mu}g/ml$, #34, #35, #38, #40 and #41 inhibited 50% cancer cell growth at the concentration of $145\;{\mu}g/ml$, $129\;{\mu}g/ml$, $173\;{\mu}g/ml$, $10\;{\mu}g/ml$ and $19\;{\mu}g/ml$ respectively. Ten of the 41 methanol extracts; #1, #3, #32, #33, #35, #36, #37, #38, #41 were active. #1 inhibited 50% cancer cell growth at the concentration of $206\;{\mu}g/ml$, #3, #32, #33, #35, #36, #37, 738, #40, #41 inhibited 50% cancer cell growth at the concentration of $133\;{\mu}g/ml$, $159\;{\mu}g/ml$, $199\;{\mu}g/ml$, $147\;{\mu}g/ml$, $113\;{\mu}g/ml$, $187\;{\mu}g/ml$, $130\;{\mu}g/ml$, $9\;{\mu}g/ml$, $15\;{\mu}g/ml$ respectively. Prescription #3, #35, #38, #40, #41 were also interesting because both methanol and water extracts were active.

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Development of Anticancer Agents from Korean Medicinal Plants. Part 9. Antitumor Evaluation of Taraxaci Herba Extracts by Colormetric Methods. (한국산 생약으로부터 항암물질의 개발 (제9보). 비색분석법에 의한 포공령 추출물의 항암평가)

  • 한두석;이명호;최규은;백승화
    • Environmental Mutagens and Carcinogens
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    • v.18 no.2
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    • pp.104-108
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    • 1998
  • In the present study, we have evaluated cytotoxic effects of Taraxaci Herba extract in human oral epitheloid carcinoma cells. An antitumor activity was measured by colorimetric assays using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) and sulforhodamine protein B (SRB). The light microscopic study showed morphological changes, Ag-NOR (argyrophylic nucleolar organizer region) number and PAS positive reaction or the treated cells. These results obtained are as follows : MTT and SRB quantities were significantly decreased in cultured KB cells treated with 10$^{-2}$ /mg/ml and 10$^{-3}$ /mg/ml concentrations. The number of Ag-NORs were significantly decreased in cultured KB cells treated with 10$^{-2}$ /mg/ml and 10$^{-3}$ /mg/ml concentrations and the rate of Ag-NORs was shifted to left side (one Ag-Nounucleus was increased and five Ag-NORs/nucleus were decreased) by the high concentration. PAS reaction of cultured KB cells treated with 10$^{-2}$ /mg/ml and 10$^{-3}$ /mg/ml concentrations was negative. These results suggest that Taraxaci Herba retains a potential antitumor activity.

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Cytotoxic activity of processing the traditional drug on monkey kidney cell (Vero) and human liver cell (WRL68) (수치(修治) 한약재가 사람의 간세포 WRL68와 원숭이의 신장세포 Vero에 미치는 영향)

  • Ju Young-Sung;Kim Ho-Kyoung;Ko Byoung-Seob
    • Journal of Society of Preventive Korean Medicine
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    • v.4 no.2
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    • pp.258-272
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    • 2000
  • The cytotoxic activities of Pinellia ternate, Aconitum carmichaeli, Arisaema amurense, Aconitum kusnezoffii and Scutellaria baicalensis on monkey kidney cell (Vero) and human liver cell (WRL68) were evaluated by Sulforhodamine B Protein (SRB) and Tetrazolium-based (MTT) colorimetric assay methods The results were as fellows : 1 The Pinellia ternate and Arisaema amurense did not show the cytotoxic activities at any concentration without processing or natural drugs. 2 The extracts of Aconitum carmichaeli, Aconitum kusnezoffii and Scutellaria baicalensis showed cytotoxic activities. However, the cytotoxic activities of processing drugs were less effective than the natural drugs. 3. The cytotoxic activities on monkey kidney cell (Vero) and human liver cell (WRL68) of Aconitum carmichaeli was determined by MTT assay. The Kyungpo(京?) of Aconitum carmichaeli showed less concentrate than that of the Dangpo(唐?). The $IC_{50}$ value on monkey kidney cell (Vero) and human liver cell (WRL68) of Kyungpo was $937{\pm}29\;and\;731{\pm}31{\mu}g/ml$, respectively 4. The cytotoxicity on monkey kidney cell (Vero) and human liver cell (WRL68) of Scutellaria baicalensis showed strong activities without processing or natural drugs.

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The Growth Inhibitory Effects of Epigallocatechin Gallate Against Human Skin Melanoma Cells and Human Oral Epitheloid Carcinoma Cells (Epigallocatechin gallate의 인체 피부흑색종세포와 인체 구강유상피암종세포에 대한 성장억제효과)

  • 한두석;박승택;백승화
    • Environmental Mutagens and Carcinogens
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    • v.18 no.2
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    • pp.98-103
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    • 1998
  • Epigallocatechin gallate (EGCG) was reported to exert weak cytotoxicity against normal healthy cells such as C3H10T1/2 cells, but profound inhibitory effects on the initiation or promotion stage of chemical carcinogenesis in mammary gland, blood and mouse skin. This study was carried out to develop antitumor agents with weak side effects and strong antitumor activity. Human skin melanoma cells (HBT 69) and human oral epitheloid carcinoma cells (OCL 17) were cultured in RPMI-1640 media containing 10% fetal bovine serum, antibiotic, and fungizone. After incubation for 24 hrs, the cells were treated with various amounts of (EGCG) for 48 hrs. The growth inhibitory effects of EGCG in human oral epitheloid carcinoma cells were evaluated by the 3- (4,5-djmethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT), neutral red (NR), and sulforhodamine B protein (SRB) assays of colorimetric methods. The light microscopic study was also carried out to observe morphological changes of the treated cells. These results obtained were as follows; 1. Significantly inhibitory effects of EGCG against cultured human oral epithelioid carcinoma cells. 2. Significantly inhibitory effects against cultured human skin melanoma cells treated with 50 $\mu$M EGCG, but decreased inhibitory effects in 100 $\mu$M EGCG. 3. Degenerative changes against cultured human oral epitheloid carcinoma cells. 4. Degenerative changes against human skin melanoma cells treated with 50 UM EGCG, but recovered degenerative changes in 100 $\mu$M EGCG.

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The Inhibitory Effects of Cannabinoids of Cannabis Against Human Skin Melanoma Cells (마자인의 카나비노이드가 인체피부흑색종세포에 미치는 억제효과)

  • Lee Ki-Nam;Kim Meung-Shin;Back Seung-Hwa
    • Journal of Society of Preventive Korean Medicine
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    • v.2 no.1
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    • pp.13-30
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    • 1998
  • Geraniol (1), olivetol (2), cannabinoids (3 and 4) and 5-fluorou.a.il (5). were tested for their growth inhibitory effects against SK-MEL-3 cell lines using two different 3-(4,5-dimethylthiazol- 2yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay and sulforhodamine B protein (SRB) assay. These compounds showed inhibitory activity in vitro in the micromolar range against SK-MEL-3 cell lines. In general, the antitumor activity of these compounds (1, 2, 3, 4 and 5) was in a dose-dependent over the micromolar concentration range $1\;to\;100{\mu}M$. The comparison of $IC_{50}$ values of these compounds in tumor cell lines shows that their susceptibility to these compounds decrease in the following order : OLVTL > CBG > CBD > 5-FU > CRNL in MTT assay, CBG > OLVTL > CBD > GRNL > 5-FU in SRB assay. Cannabinoids (3 and 4) and 5-fluorouracil (5) were tested for their cytotoxic effects on NIH 373 fibroblasts using two different MTT assay and SRB assay. These compounds exhibited potent cytotoxic activities in vitro in the micromolar range against NIH 373 fibroblasts. In general, the cytotoxic activities of these compounds (3, 4 and 5) were in a dose-dependent over the micromolar concentration range $1\;to\;100{\mu}M$. The comparison of $CD_{50}$ values of these compounds on NIH 373 fibroblasts shows that their susceptibility to these compounds decrease on the following order ; CBD > 5-FU > CBG in MTT assay and SRB assay. Cannabigerol (3) was shown the least cytotoxic activity on NIH 373 fibroblasts. Cannabigerol (3) exhibited the most growth-inhibitory activity against SK-MEL-3 cell lines.

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The Protective Effects of the Extract of Saururus chinesis against Cadmium Induced Cytotoxicity(II) (삼백초 추출물의 카드뮴독성에 대한 방어효과(II))

  • 이정호;유일수;이기남;지정목;한두석;신민교;정승일;오현주;백승화
    • Toxicological Research
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    • v.17 no.3
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    • pp.173-180
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    • 2001
  • This study was conducted to investigate the antitoxic component in ethanol extract of Saururus chinesis (S. chinesis). The results were as follows: Generally, detoxication effects by s. chinesis extract increased in proportion to the extract concentration. non 8 $\mu\textrm{g}$/g dosage of S. chinesis extract was administered, it showed the highest antitoxic effects in metallothionein induction. After the extract treatment, body weights generally increased In proportion to the extract concentrations. from the above results, S. chinesis extract Increased Metallothionein concentration and decreased the toxicity of cadmium In rats. In vitro the antitoxic activity of ethanol extract of S. chinesis on NIH3T3 fibroblasts was evaluated by the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide) and SRB (sulforhodamine B protein) assays. The light microscopic study was carried out to observe morphological changes of the treeated cells. $10^{-2}$mg/ml Concentrations of S. chinesis extract was shown significant antitoxic activity. The number of NIH 3T3 fibroblasts were increased and tend to regenerate. These result suggest that S. chinesis extract retains a potential antitoxic activity.

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The Inhibitory Effects of Houttuynia cordata $T_{HUNB}$ against Cadmium induced Cytotoxicity (II) (어성초의 카드뮴에 대한 독성억제효과 (II))

  • 이정호;유일수;이기남;정우영;한두석;백승화
    • YAKHAK HOEJI
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    • v.44 no.5
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    • pp.432-439
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    • 2000
  • This study was condo더ed to investigate the antitoxic component in aqueous extract of Houttuynia cordata $T_{HUNB}$. The results were as follows: Generally, detoxification effects by Houttuynia cordata $T_{HUNB}$ extract increased in proportion to the extract concentrations in rats. When 40 mBtg dosage of Houttuynia cordata $T_{HUNB}$ extract was administrated, Houttynia cordata $T_{HUNB}$ extract showed the highest antitoxic effects in metallothionein induction. After the extract treatment, body weights increased in proportion to the extract concentrations. However, after 3 weeks, the body weight decreased insignificantly. From the above results, Houttuynia cordate $T_{HUNB}$ extract increased metallothionein concentration and decreased the toxicity of cadmium in rats. In vitro the antitoxic activity of aqueous extract of Houttuynia cordata $T_{HUNB}$ on NIH 373 fibroblasts was evaluated by the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetra-zoliumbromide) and SRB (sulforhodamine B protein) assays. The light microscopic study was carried out to observe morphological changes of the treated cells. These results were obtained as follows; The concentration of 10$^{-2}$ mg/ml of Houttuynia cordata $T_{HUNB}$ extract was shown significant antitoxic activity The number of NIH 373 fibroblasts were increased and tend to regenerate. These results suggest that Houttuynia cordata THUNB extract retains a potential antitoxic activity.oxic activity.

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Cytotoxicity of Crude Extracts of Rheum uudulatum L. with Human Kidney Epithelial Cell A498 (신장 상피세포주 A498을 이용한 대황(Rheum undulatum L.)추출물의 세포독성)

  • 나명석;진종언;조남철
    • The Korean Journal of Food And Nutrition
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    • v.13 no.5
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    • pp.460-464
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    • 2000
  • We have evaluated cytotoxic effects of four crude extracts of methylene chloride, ethyl acetate, butanol, water layer isolated Rheum undulatum in A498 cell line, human kidney epithelial cells. The cytotoxic evalutation was measured by colorimetric assay using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide(MTT) , neutral red(NR) and sulforhodamine protein B(SRB). These results obtained are as follows : MTT, NR and SRB quantities were significantly decreased in cultured A498 cells treated four crude extracts by increased concentrations. The cell cytotoxic effect of crude extracts of butanol layer was more stronger than others layer. The values of MTT$\sub$50/, NR$\sub$50/, SRB$\sub$50/ of crude extract of butanol layer and were measured both 0.63 mg/ml, 0.65 mg/ml, and 0.68 mg/ml, respectively and the values of water layer were 0.84 mg/ml, 0.82 mg/ml. and 0.80 mg/ml. respectively in cultured A498 cell line.

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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.

Effect of Various Factors on Early THP-1 Cell Adhesion Induced Phorbol 12-Myristate 13-Acetate (PMA) (Phorbol 12-myristate 13-acetate (PMA) 처리로 유도되는 THP-1 세포의 초기 부착에 관한 다양한 인자의 효과)

  • Jo, Yong-Sam;Shin, Ji-Hyun;Choi, Tae-Saeng
    • Journal of Life Science
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    • v.18 no.7
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    • pp.952-957
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    • 2008
  • We evaluated the effects of various factors (e.g., serum, inhibitors of protein synthesis, and cytoskeleton and protein kinases) on early PMA-induced THP-1 cell adhesion using an adhesion assay with Sulforhodamine B (SRB) staining, which was used to assess the proliferation of the attached cells. THP-1 cell adhesion to a plastic substrate was detected 1 hr after exposure to Phorbol 12-Myristate 13-Acetate (PMA) and peaked after 18 hr. At concentrations > 25 nM PMA, the level of adhesion did not change. Based on our preliminary results, we used 25 nM PMA and 5 hr of culture as standard assay conditions. Early PMA-induced cell adhesion was not affected by the presence of serum or PD 98059 in the culture medium, but was affected by the addition of PKC inhibitors and cycloheximide. In the presence of actin inhibitor with PMA, the cell adhesion increased when comparing with PMA treatment only. Thus, early PMA-induced adhesion of THP-1 cells does not require serum in the culture medium, MAP-kinase activation, or actin polymerization, but does require de novo protein synthesis and PKC activation. Our SRB-based cell adhesion assay may be used to screen other PKC inhibitors.