• Title/Summary/Keyword: SRB assay

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

  • Lee, Byung-Cheol;Choi, Sang-Ho;Kim, Tae-Sung
    • Journal of Microbiology and Biotechnology
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    • v.14 no.2
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    • pp.350-355
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    • 2004
  • Sulforhodamine B (SRB) assay is a rapid, sensitive, and inexpensive method for measuring cell proliferation and chemosensitivity. However, the lactate dehydrogenase (LDH) release assay is generally used to measure cytototoxicity of infectious microorganisms against host cells. In this study, we investigated the possibility of applying the SRB assay to determine cytotoxicity for infectious microorganisms, and compared the results with those obtained by the LDH release assay. We used Vibrio vulnificus as a model of infectious microorganisms. The SRB assay showed that V vulnificus strongly induced cytotoxic activity against human intestinal cells, Caco-2 and INT-407 cells. The degree of cytotoxicity closely correlated with infection time and number ratios of V. vulnificus to intestinal cells (MOI, multiplicity of infection). Furthermore, cytotoxicity values obtained by SRB assay correlated well with results obtained by the LDH release assay, and both assays gave a linear response with respect to MOI Heat-inactivation of V. vulnificus for 35 min at $60^{\circ}C$ did not induce cytotoxic activity, indicating that viability of V. vulnificus is crucial for cytotoxic activity against intestinal cells. Although both assays are suitable as cytotoxicity endpoints, the SRB assay is recommended for measuring cytotoxicity of infectious microorganisms against host cells because of its significantly lower cost and more stable endpoint than the LDH release assay.

Anticancer Effect of Doenjang in in vitro Sulforhodamine B (SRB) Assay (된장의 in vitro Sulforhodamine B (SRB) Assay에 의한 암세포 증식 억제 효과)

  • 이숙희;임선영;박건영
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.28 no.1
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    • pp.240-245
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    • 1999
  • Growth inhibitory effect of doenjang(Korean soypaste) methanol extracts in SRB assay using AGS human gastric adenocarcinoma cell, Hep 3B human hepatocellular carcinoma cell and HT 29 human colon cancer cell was studied. The treatment of doenjang methanol extracts(2mg/assay) to the AGS, Hep 3B and HT 29 cancer cells inhibited the growth of the cancer cells by 55%, 60%, and 71%, respectively. Doenjang methanol extracts exhibited the highest inhibitory effect among other soybean fermented foods and original materials in the SRB assay. In addition, to separate active compounds of doenjang methanol extracts, we fractionated the doenjang with hexane, methanol, dichloromethane, ethylacetate and butanol. Growth inhibitory effect on the AGS, Hep 3B, HT 29 and MG 63 cancer cells was the highest in the fractions of dichloromethane and ethylacetate among other solvent fractions of the doenjang. These results showed that some compounds contained in the fractions of dichloromethane and ethylacetate might play a role on the anticanceric effect of doenjang.

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전남지역 생활하수처리장과 농공단지폐수처리장 방류수에 대한 세포독성 평가 - XTT, SRB in vitro bio-assay를 이용한 -

  • Lee, Moon-Hee;Han, Sang-Guk
    • Proceedings of the Korean Environmental Sciences Society Conference
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    • 2005.11a
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    • pp.185-188
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    • 2005
  • XTT assay와 SRB assay를 이용하여 Hep3B와 L929 세포에서 생활하수처리장과 농공단지폐수처리장 방류수의 세포독성을 알아본 결과 다음과 같은 결론을 얻었다. 1) XTT, SRB 실험법에 적용하여 대표적인 발암성 물질인 Benzo(a)Pyrene의 세포독성을 평가 할 수 있었다. Benzo(a)Pyrene 30uM농도는 Hep 3B와 L929 세포에서 약 50%의 세포독성을 나타내었다. 2) 단백질합성과 세포호흡의 활성도로 세포독성을 평가할 수 있는 실험법인 XTT와 SRB 실험법에 적용하여 방류수를 추출 농축한 실제 시료에 대한 세포독성을 평가할 수 있었다. 3) 생활하수처리장의 방류수에 비해 다종 고농도의 화학물질로 오염되어있는 농공단지폐수처리장 방류수에서의 독성발현이 더 높았다. 4) 생활하수처리장 방류수와 농공단지폐수처리장 방류수 중 오염도가 가장 심각한 4지점의 화학물질 분석 결과와 in vitro bio-assay에 의한 세포독성 결과가 일치하였다.

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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
    • Advances in Traditional Medicine
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    • v.1 no.1
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    • pp.45-54
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    • 2000
  • 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.

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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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Anticancer Effects of Organic Chinese Cabbage Kimchi

  • Park, Woon-Young;Park, Kun-Young
    • Preventive Nutrition and Food Science
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    • v.4 no.2
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    • pp.113-116
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    • 1999
  • The anticancer effect of methanol extracts from common Chinese cabbage kimchi(CC kimchi ) and organically cultivated Chinese cabbage kimchi (OC kimchi) was studied on the cell growth, MTT assay and SRB assay using AGS human gastric cancer cells. Methanol extracts from CC kimchi and OC kimchi exhibited the anticancer activites in vitro and in vivo. Methanol extract from 6 day-fermented CC kimchi and OC kimchi inhibited the growth of AGS cells by 55.2 and 60.7% , respectively. At MTT assay an dSRB assay, 6 day-fermented OC kimchi showed higher inhibition rate (MTT : 42%, SRB : 61%) than 6 day-fermented CC kimchi(MTT : 33%, SRB : 52%). Methanol extracts from 6-day fermented CC kimchi and OC reduced the tumor formation and prolonged the life span of sarcoma-180 cell injected Balb.c mouse. OC kimchi treated group resulted in the smaller tumor weight of 4.58$\pm$0.32g compared th the CC kimchi group of 5.40$\pm$0.78g and the control group of 7.50$\pm$0.54g and OC kimchi treted group (25.3 days) lived longest among control (20.2days ) and CC kimchi(23.5days) treted groups.

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Synthesis and Cytotoxic Effects of Deoxy-tomentellin

  • Han, Du-Seok;Jung, Kui-Ho;Jung, Woo-Jung;Oh, In-Kyo;Kang, Kil-Ung;Baek, Seung-Hwa
    • Archives of Pharmacal Research
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    • v.23 no.2
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    • pp.121-127
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    • 2000
  • Cannabigerol (1, CBG), methyl 4-[(2E)-3,7-dimethyl-2,6-octad ienyl)oxy]-3-methoxybenzoate (2, DTM), 5-fluorouracil (3, FU) as a reference, and cannabidiol (4, CBD) were tested for their growth inhibitory effects against KB(ATCC NO, OCL 17) cell lines using two different assays, the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazoliumbromide (MTT) assay and the sulforhod-amine B protein (SRB) assay. These compounds showed inhibitory activity in vitro in the micromolar range against KB cell lines. In general, the antitumor activities of these compounds (1, 2, 3 and 4) were dose-dependent over the micromolar concentration range of 1 to 100 M. The comparison of $IC_{50}$ values of these compounds in tumor cell lines showed that their susceptibility to these compounds decreases in the following order: DTM > CBD > 5-FU > CBG by MTT assay and DTM = CBD > 5-FU > CBG by SRB assay. CBG 1, DTM 2, 5-FU 3, and CBD 4 were tested for their cytotoxic effects on NIH 3T3 fibroblasts using two different assays, the 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 acivities of these compounds (1, 2, 3 and 4) were dose-dependent over the micromolar concentraion range of 1 to 100 M. The comparison of $CD_{50}$ values of these compounds in NIH 3T3 fibroblasts shows that their susceptibility to these compounds in decreases the following order(:) CBD > 5-FU > DTM > CBG by MTT assay, CBD > 5-FU > CBG > DTM by SRB assay. These results suggest that DTM 2 has the most growth-inhibitory activity against KB cell lines.

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Antitumor Evaluation of Cannabidiol and Its Derivatives by Colorimetric Methods

  • Baek, Seung-Hwa;Shin, Ji-Hee;Chung, Woo-Young;Han, Du-Seok
    • Toxicological Research
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    • v.16 no.2
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    • pp.101-107
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    • 2000
  • 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.

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Screening and Isolation of the Antitumor Agents from Medicinal Plants (I) (생약으로부터 항암성분의 검색 및 분리 (I))

  • Park, Shin-Young;Kim, Jin-Woong
    • Korean Journal of Pharmacognosy
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    • v.23 no.4
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    • pp.264-267
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    • 1992
  • The cytotoxic activity of medicinal plants was screened using A549 human lung cancer cell line. Plant materials were extracted with 80% methanol and fractionated to chloroform and water layers. Each methanol, chloroform, and water extract of thirty-two medicinal plants was tested for cytotoxic activity in A549 cell culture system and the cell viability was measured by SRB assay.

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Antioxidant Activity and Cytotoxic Effect of an Ethanol Extract from Seoritae (서리태 에탄올 추출물의 항산화 활성 및 암세포 증식 억제 효과)

  • Jeon, Yeon-Hee;Won, Ji-Hye;Kwon, Ji-Eun;Kim, Mee-Ra
    • Korean journal of food and cookery science
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    • v.27 no.3
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    • pp.1-10
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    • 2011
  • The antioxidant activity and cytotoxic effect of an ethanol extract from Seoritae were analyzed to develop new functional food materials. The antioxidant activity of Seoritae was determined by measuring electron donating ability with 1,1-diphenyl-2picrylhydrazyl (DPPH) and 2-2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) assays, as well as the ferric reducing antioxidant power (FRAP) assay. The cytotoxic effect of the Seoritae ethanol extract was measured with the 3-(4,5-dimethylthiazol-2-yl)-2,5-dipheltetrazolium (MTT) and sulforhodamine B (SRB) assays. As a result, the electron donating abilities of Seoritae against the DPPH and ABTS radicals were 63.75% and 87.68% at 500 ${\mu}g$/assay, respectively. The $IC_{50}$ values of Seoritae in the DPPH and ABTS assays were 385.39 ${\mu}g$/assay (128.46 ${\mu}g/mL$) and 209.39 ${\mu}g$/assay (51.83 ${\mu}g/mL$). Additionally, the FRAP value of Seoritae was 0.84 $FeSO_4$ eq. mM at 800 ${\mu}g$/assay. The total amounts of polyphenols and flavonoids, which indicate the antioxidant capability of Seoritae extract were 1.65 mg/g and 0.59 mg/g, respectively. Moreover, Seoritae extract showed a high cytotoxic effect of up to 81% against human cancer cells, particularly A-549 and HeLa cells. The growth inhibition rate of Seoritae extract against A-549 and HeLa cells was up to 76.48% and 75.67% in the MTT assay, and 78.98% and 80.54% in the SRB assay, respectively. The results of this study suggest that an ethanol extract of Seoritae is a potentially good natural antioxidant.