Sulforhodamine B Assay to Determine Cytotoxicity of Vibrio vulnificus Against Human Intestinal Cells

( Lee Byung Cheol ),( Sang Ho Choi ),( Tae Sung Kim ) 한국미생물생명공학회 2004 Journal of microbiology and biotechnology Vol.14 No.2

Development of in vitro Evaluation Model in Dental Biocompatibility by Non-radioactive Materials

Lee, Dong-Keun,Lee, Yong-Keun,Yoo, Soo-Kyoung,Kim, Kang-Ju,Lee, Heung-Soo,Lee, Kwang-Hee,Han, Doo-Seok The Official Publication of Korean Academy of Oral 1995 International Journal of Oral Biology Vol.19 No.2

The purpose of this study was 1)to investigate the cytotoxicity of some dental materials in cultures with or without elution 2)to determine the effect of serum concentration on the measurement of cytotoxicity 3)to evaluate the difference of sensitivity of MTT assay and SRB assay methods. Composite resin(COM), Miracle mix??(MIM), polycarboxylate cement(PCA), and zinc phosphate cement(ZPC) were investigated. The materials were hardened according to the manufacturer's directions and the leachable components were dissolved in a 37℃ shaking water bath for 24 hours in one group while another group was not leached. Fibroblasts were derived from healthy human gingiva. Fibroblasts between the 5th and 8th passages were used in this study. The culture medium was α-MEM, supplemented with 10% or 1% FBS. In the MTT assay, 0.1 mg of MTT was added to each of 24 wells 24 hours after the specimens immersed in the culture. After being removed from the medium, 200 ㎕ of DMSO was added to each well. The spectrophotometric absorbance at 570 nm was then measured by an ELISA reader. In the SRB assay, 10% of cold TCA solution was added to each well 24 hours after the specimens were immersed in the culture. Next 500 ㎕ of 0.4% SRB solution was added and stained for 30 minutes, and the dye was eluted with 10 mM Tris. The spectrophotometric absorbance at 570 nm was then measured by an ELISA reader. In the SRB assay, 10% of cold TCA solution was added to each well 24 hours after the specimens were immersed in the culture. Next 500 ㎕ of 0.4% SRB solution was added and stained for 30 minutes, and the dye was eluted with 10 mM Tris. The spectrophotometric absorbance at 520 nm was then measured by an ELISA reader. Both the MTT and the SRB assay showed that the cytotoxicity of dental materials in 1%-serum culture was higher than that in 10% serum condition. After eluting the specimens, the cytotoxicity of MIM and ZPC in the eluted group was higher than that of the non-eluted group. There was no significant difference in the case of COM and PCA. In conclusion, the measuring a cytotoxicity in a 1%-serum culture provided more sensitive results than that in 10 %-serum culture, and the SRB assay showed more sensitive and reproducible results than MTT assay.

SRB Assay에 의한 수수 분획물의 in Vitro 항암효과

권영미,김보경,이숙희,박건영 대한암예방학회 2007 Journal of cancer prevention Vol.12 No.2

Anticancer effects of various cereals were studied by surforhodamine B (SRB) assay on AGS human gastric adenocarcinoma cells and HT-29 human colon carcinoma cells. The methanol extracts of sorghum, millet and Job's tears showed more than 50% of inhibition rate on the AGS cells and the HT-29 cells. Among the cereals, sorghum showed the highest inhibition rate on both the cancer cells. To separate and identify the anticancer compounds from sorghum, the methanol extract and five fractions (fr.) (hexane fr., dichloromethane fr., ethylacetate fr., butanol fr. and aqueous fr.) were prepared, and SRB assay on the AGS cells and the HT-29 cells were examined. The dichloromethane fr. and ethylacetate fr. of the sorghum exhibited strong cytotoxicity effects than other fractionated samples. The sorghum extract and fractions inhibited survival of AGS cells and HT-29 cells in SRB assay. Among the fractions, dichloromethane fr. and ethylacetate fr. of the sorghum showed strong inhibitory effect on both AGS cells and HT-29 cells as concentration dependent-manner. Especially, dichloromethane fr. and ethylacetate fr. strongly inhibited the growth of AGS cells. This study resulted in sorghum, especially compounds in dichlromethane and ehtylacetate fractions exhihibited higest anticancer effect among the tested cereals. (Cancer Prev Res 12, 136-142, 2007)

The antiproliferative activity of cannabidiol ethyl ethers against human ora epitheloid carcinoma cells

Baek, Seung-Hwa,Kang, Kil-Ung,Chung, Soon-Ryang,Kim, Hyung-Min,Chung, Woo-Young,Han, Du-Seok Kyung Hee Oriental Medicine Research Center 2000 International journal of oriental medicine Vol.1 No.1

Cannabidiol derivatives (1, 2 and 3), and 5-fluorouracil (4, 5-FU) were tested for their growth inhibitory effects against human oral epitheloid carcinoma cell lines (KB) using two different 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay and sulforhodamine B protein (SRB) assay. These compounds showed a potent inhibitory activity in vitro in the micromolar range against KB cell lines. In general, the antitumor activity of these compounds (1, 2, 3 and 4) was in a dose-dependent over the micromolar concentration ranges from $1\;{\mu}M\;to\;100\;{\mu}M$. The comparison of $IC_{50}$ values of these compounds in tumor cell lines shows that their susceptibility to these compounds decreases in the following order: CBD > 5-FU > CBDME > CBDDE by the MTT assay and SRB assay. Cannabidiol derivatives (1, 2 and 3), and 5-FU were tested for their cytotoxic effects on NIH 3T3 fibroblasts using two different MTT assay and SRB assay. These compounds exhibited potent cytotoxic activities in vitro in the micromolar range against NIH 3T3 fibroblasts. In general, the cytotoxic activities of these compounds (1, 2, 3 and 4) were in a dose-dependent over the micromolar concentration range $1\;{\mu}M\;to\;100\;{\mu}M$. The comparison of $CD_{50}$ values of these compounds on NIH 3T3 fibroblasts shows that their susceptibility to these compounds decreases in the following order; CBD > 5-FU > CBDDE > CBDME by MTT assay, CBD > 5-FU > CBDME > CBDDE by SRB assay. These results suggest that cannabidiol (1, CBD) retains the most growth-inhibitory activity against KB cell lines.

백강잠 전탕액이 Hydrogen Peroxide에 의해 손상된 배양 척수운동신경세포에 미치는 효과

이민주,이영보,최규선,신병철,권영달,송용선 대한동의생리학회,대한동의병리학회 2001 동의생리병리학회지 Vol.15 No.6

To evaluate the effect of Batryticatus Bombycis(BB) water extract on cultured mouse spinal motor neuron which was inhibited by hydrogen peroxide (H_2O_2)-induced oxygen radicals, MTT assay, NR assay, TBARS assay and SRB assay were carried out after the cultured mouse spinal motor neuron were preincubated with various concentrations of BB water extract for 3 hours prior to exposure of H_2O_2. H_2O_2, a oxygen radical, decreased the survival rate of the cultured mouse spinal motor neuron cells on NR assay and MTT assay. BB water extract have efficacy of decreasing a amount of lipid peroxidation increased by H_2O_2 in mouse spinal motor neuron and increasing total protein synthesis activity decreased by H_2O_2 in mouse spinal motor neuron. From the above results, It is concluded that BB has marked efficacy as a treatment for the damages caused in the H_2O_2-mediated oxidative process.

메주 메탄올 추출물 및 분획물의 항돌연변이 및 인체 암세포 성장 억제 효과

임선영,박건영,이숙희,최재수 한국생명과학회 2007 생명과학회지 Vol.17 No.2

Inhibitory effects of methanol extracts and several solvent fractions from meju on mutagenicity in vitro genotoxicity (SOS chromotest) and growth of human cancer cells (AGS gastric adenocarcinoma and Hep 3B hepatocellular cancinoma cells) were studied. The treatment of meju methanol extracts (100 μg/assay) to SOS chromotest system inhibited N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) induced mutagenicity by 36%. However, the ethylacetate and dichloromethane fractions from meju methanol extracts showed the stronger antimutagenic effects (91% and 91%, respectively) in SOS chromotest. In sulforhodamine B (SRB) assay, the treatments of ethylacetate and dichloromethane fractions (2 mg/assay) significantly inhibited the growth of AGS and Hep 3B cancer cells by 64% and 71%, respectively. These results indicated that meju had inhibitory effects on MNNG in SOS mutagenic system and growth of human cancer cells, suggesting that its antimutagenic effect may be relative to activity of doenjang. 메주의 헥산 및 메탄올 추출물과 그것을 용매로 분획하여 얻어진 디클로로메탄, 에틸아세테이트, 부탄올, 물 분획물들에 의한 SOS chromotest 실험계에서 항돌연변이 효과 및 SRB assay를 이용한 인체 암세포들의 성장 억제 효과에 대하여 검토하였다. SOS chromotest 실험계의 경우, 첨가농도 100 μg/assay에서 메주의 디클로로메탄 및 에틸아세테이트 분획물은 각각 91%, 91%로 MNNG에 대하여 강한 항돌연변이 효과를 나타났다. AGS 인체 위암세포와 Hep 3B 인체 간암세포를 이용하여 항암효과를 실험한 결과, 메주 에틸아세테이트분획물은 첨가농도 200 μg/assay에서 각각 93% 및 91%로 암세포 성장을 크게 저해시켰고 그 저해 효과는 디클롤메탄 분획물보다 더 높았다. 인체 위암세포인 AGS를 이용한 SRB assay을 이용한 억제 효과 실험에서 메주의 에틸아세테이트 분획물은 첨가농도 2 mg/assay에서 인체 위암 및 간암세포의 증식을 각각 64% 및 71%로 억제하여 분획물들 중 가장 높은 저해효과를 보였고 헥산 및 디클로로메탄 분획물은 그 다음으로 억제 효과가 높았다. 이러한 결과는 된장 메탄올 추출물과 그 분획물의 결과와 유사한 경향을 보였으므로 메주와 된장이 여러 종류의 미생물, 곰팡이류와 세균류들에 의해 발효과정을 거치는 동안 원재료인 콩에서는 없었던 혹은 함량이 적은 성분들이 생성되거나 증가되어 항암효과를 나타내는 것으로 추정되어 진다.

메주 메탄올 추출물 및 분획물의 항돌연변이 및 인체 암세포 성장 억제 효과

임선영,박건영,이숙희,최재수,Lim, Sun-Young,Park, Kun-Young,Lee, Sook-Hee,Choi, Jae-Soo 한국생명과학회 2007 생명과학회지 Vol.17 No.1

메주의 헥산 및 메탄올 추출물과 그것을 용매로 분획하여 얻어진 디클로로메탄, 에틸아세테이트, 부탄올, 물 분획물들에 의한 SOS chromotest 실험계에서 항돌연변이 효과 및 SRB assay를 이용한 인체 암세포들의 성장 억제 효과에 대하여 검토하였다. SOS chromotest 실험계의 경우, 첨가농도 $100{\mu}g/assay$에서 메주의 디클로로메탄 및 에틸아세테이트 분획물은 각각 91%, 91%로 MNNG에 대하여 강한 항돌연변이 효과를 나타났다. AGS 인체 위암세포와 Hep 3B 인체 간암세포를 이용하여 항암효과를 실험한 결과, 메주 에틸아세테이트분획물은 첨가농도 $200{\mu}g/assay$에서 각각 93% 및 91%로 암세포 성장을 크게 저해시켰고 그 저해 효과는 디클롤메탄 분획물보다 더 높았다. 인체 위암세포인 AGS를 이용한 SRB assay을 이용한 억제 효과 실험에서 메주의 에틸아세테이트 분획물은 첨가농도 2 mg/assay에서 인체 위암 및 간암세포의 증식을 각각 64% 및 71%로 억제하여 분회물들 중 가장 높은 저해효과를 보였고 헥산 및 디클로로메탄 분획물은 그 다음으로 억제 효과가 높았다. 이러한 결과는 된장 메탄올 추출물과 그 분획물의 결과와 유사한 경향을 보였으므로 메주와 된장이 여러 종류의 미생물, 곰팡이류와 세균류들에 의해 발효과정을 거치는 동안 원재료인 콩에서는 없었던 혹은 함량이 적은 성분들이 생성되거나 증가되어 항암효과를 나타내는 것으로 추정되어 진다. Inhibitory effects of methanol extracts and several solvent fractions from meju on mutagenicity in vitro genotoxicity (SOS chromotest) and growth of human cancer cells (AGS gastric adenocarcinoma and Hep 3B hepatocellular cancinoma cells) were studied. The treatment of meju methanol extracts $(100{\mu}g/assay)$ to SOS chromotest system inhibited N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) induced mutagenicity by 36%. However, the ethylacetate and dichloromethane fractions from meju methanol extracts showed the stronger antimutagenic effects (91% and 91%, respectively) in SOS chromotest. In sulforhodamine B (SRB) assay, the treatments of ethylacetate and dichloromethane fractions (2 mg/assay) significantly inhibited the growth of AGS and Hep 3B cancer cells by 64% and 71%, respectively. These results indicated that meju had inhibitor)r effects on MNNG in SOS mutagenic system and growth of human cancer cells, suggesting that its antimutagenic effect may be relative to activity of doenjang.

한국산 생약으로 부터 항암물질의 개발 (제7보), 소엽의 Chloroform 가용성 분획이 인체 구강유상피암종세포에 미치는 세포독성작용

한두석,김영일,최규은,곽정숙,백승화,Han, Du-Seok,Kim, Young-Il,Choi, Kyw-Eun,Kwag, Jung-Suk,Baek, Seung-Hwa 한국환경성돌연변이발암원학회 1998 한국환경성돌연변이·발암원학회지 Vol.18 No.1

In the present study, we have evaluated cytotoxic effects of the chloroform soluble fraction of the methanolic extract of Perilla frutescens in human oral epitheloid carcinoma cells. The light microscopic study showed morphological changes of the treated cells. Cell membrane damaging activity was measured by the lactate dehydrogenase (LDH) assay and disruptions in cell organelles were determined by 3-(4,5-dime-thylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT), neutral red (NR) and sulforhodamine protein B (SRB) of colorimetric assay. These results suggest that Perilla frutescens retains a potential antitumor activity.

Antitumor Evaluation of Cannabidiol and Its Derivatives by Colorimetric Methods

Baek, Seung-Hwa,Shin, Ji-Hee,Chung, Woo-Young,Han, Du-Seok The Korean Society of Toxicology Korea Environment 2000 Toxicological Research Vol.16 No.2

Cannabidiol derivatives (1, 2 and 3), 5-fluorouracil (4, 5-FU) and adriamycin (5, AM) were tested for their growth inhibitory effects against human tumor cell lines using two different 3-{4,5-dimeth-ylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay and sulforhodamine B protein (SRB) assay. The light microscopic study showed morphological changes of the treated cells. Disruptions in cell organelles were determined by colorimetric methods; MTT assay and STB assay. These results suggest that cannabidiol (1, CBD) retains the most growth-inhibitory activity against human tumor cell lines.

한국산 생약으로부터 항암물질의 개발(제 8보) - 포공령 추출물이 인체 피부흑색종세포에 미치는 세포독성작용 -

오인교,유은아,한두석,강길웅,백승화,Oh, In-Kio,Yoo, Eun-Ah,Han, Du-Seok,Kang, Kil-Ung,Baek, Seung-Hwa 한국생약학회 1998 생약학회지 Vol.29 No.3

In the present study, we have evaluated cytotoxic effects of Taraxaci herba extract on human skin melanoma cells. The light microscopic study showed morphological changes of the treated cells. Disruptions in cell organelles were determined by calorimetric methods: MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazoliumbromide), NR (Neutral red) and SRB (Sulforhodamine B protein) assay. These results suggest that Taraxaci herba retains a potential antitumor activity.