• Title/Summary/Keyword: MTT assay

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Selective Cytotoxicity of Novel Platinum(II) Coordination Complexes Containing DL-2-Hydroxy 3-Methylbutyric Acid (DL-2 하이드록시 3-메틸 부틸산물 배위자로 한 새로운 항암성 백금(II) 착체의 위암세포와 정상신장세포에 대한 선택적 세포독성)

  • 정지창;홍언표;최승기;장성구;육창수;노영수
    • Biomolecules & Therapeutics
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    • v.11 no.2
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    • pp.91-98
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    • 2003
  • A new series of highly water soluble platinum(II) complexes[Pt(II)(DL-2-hydroxy-3-methylbutyrate)(trans-l-1,2-dimninocyc1ohexane)] (PC-1) and [Pt(II)DL-2-hydroxy-3-methylbutyrate](cis-1,2-diaminocyclohexane)](PC-2) were synthesized and characterized by their elemental analysis and by various spectroscopic techniques [infrared(IR), $^{13}C$-nuclear magnetic resonance (NMR)]. In vitro antitumor activity of new Pt(II)complexes was tested against MKN-45, MKN/ADM and MKN/CDDP human gastric adenocarcinoma cell lines using colorimetric MTT[3-(4,5-dimethyl thiazol-2-yl)-2,5-diphenyltetrazoliumbromide] assay for cell survival and proliferation. PC-1 and PC-2 showed active against MKN-45/P, MKN/ADM and MKN/CDDP human gastric cancer cell lines, and the antitumor activity of these compounds were comparable or superior to that of cisplatin. The nephrotoxicities of PC-1 and PC-2 were found quite less then that of cisplatin using MTT and [$^3H$] thymidine uptake tests in rabbit proximal tubule cells, human kidney cortical cells human renal cortical tissues. Based on these results, these novel platinum(II) complex compounds(PC-1 & PC-2) represent a valuable lead in the development of the new anticancer chemotherapeutic agents capable of improving antitumor activity and low nephrotoxicity.

Effects of He-Ne Laser Irradiation on the Activity of the Cultured Fibroblast (He-Ne 레이저 조사가 배양 섬유모세포의 활성에 미치는 영향)

  • Song In-Young;Lee Jae-Hyoung
    • The Journal of Korean Physical Therapy
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    • v.9 no.1
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    • pp.71-79
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    • 1997
  • The purpose of this study were to determined the effect of laser irradiation on the fibroblast activity. Cultures of 3T3 fibroblasts were subjected to Helium Neon laser(632.8 nm) irradiation of various energy density. On one, two and three consecutive days the fibroblast monolayers wert irradiated for period from 0 to 32 minutes with 8 mW of average output power. The fibroblast activity was determined by the quantitative assay of MTT, SRB and NR after incubation of the fibroblasts for 24 hours. Results show that exposure duration from 2 min to 32 min could increase MTT at three consecutive days, whereas control and 1 min, one and two days irradiation had were not inclosed. The SRB and NR were inclosed at two and three consecutive days from 2 min to 32 min, whereas control and 1 min, and once radiation were not increased. These result demonstrate that energy density from 0.48 to 7.64 J/cm could increase cellular protein contents and fibroblast activity at more than twice irradiation of laser, whereas low energy density (less than 0.24 J/m) and once irradiation of laser had no effect. The results suggest that the beneficial effect of the He-Ne laser with adequate dose on fibroblast activity in vitro.

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Identification of Novel Bioactive Hexapeptides Against Phytopathogenic Bacteria Through Rapid Screening of a Synthetic Combinatorial Library

  • Choi, Jae-Hyuk;Moon, Eun-Pyo
    • Journal of Microbiology and Biotechnology
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    • v.19 no.8
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    • pp.792-802
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    • 2009
  • Antimicrobial peptides (AMPs) are considered to be a promising alternative to conventional antibiotics for future generations. We identified four novel hexapeptides with antimicrobial activity: KCM11 (TWWRWW-$NH_2$), KCM12 (KWRWlW-$NH_2$), KCM21 (KWWWRW-$NH_2$), and KRS22 (WRWFIH-$NH_2$), through positional scanning of a synthetic peptide combinatorial library (PS-SCL). The ability of these peptides to inhibit the growth of a variety of bacteria and unicellular fungi was evaluated. KCM11 and KRS22 preferentially inhibited the normal growth of fungal strains, whereas KCM12 and KCM21 were more active against bacterial strains. Bactericidal activity was addressed in a clear zone assay against phytopathogenic bacteria, including Pectobacterium spp., Xanthomonas spp., Pseudomonas spp., etc. KCM21 showed the highest activity and was effective against a wide range of target organisms. Application of KCM21 with inoculation of Pectobacterium carotovorum subsp. carotovorum on detached cabbage leaves resulted in an immune phenotype or a significant reduction in symptom development, depending on the peptide concentration. Cytotoxicity of the four hexapeptides was evaluated in mouse and human epithelial cell lines using an MTT test. The results revealed a lack of cytotoxic effects.

Cytotoxic Activity and Three-Dimensional Quantitative Structure Activity Relationship of 2-Aryl-1,8-naphthyridin-4-ones

  • Kim, Yong-Jin;Kim, Eun-Ae;Chung, Mi-Lyang;Im, Chae-Uk
    • The Korean Journal of Physiology and Pharmacology
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    • v.13 no.6
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    • pp.511-516
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    • 2009
  • A series of substituted 2-arylnaphthyridin-4-one analogues, which were previously synthesized in our laboratory, were evaluated for their in vitro cytotoxic activity against human lung cancer A549 and human renal cancer Caki-2 cells using MTT assay. Some compounds (11, 12, and 13) showed stronger cytotoxicity than colchicine against both tumor cell lines, and compound 13 exhibited the most potent activity with $IC_{50}$ values of 2.3 and $13.4\;{\mu}M$, respectively. Three-dimensional quantitative structure activity relationship (3D-QSAR) studies of comparative molecular field analysis (CoMFA) and comparative molecular similarity indices analysis (CoMSIA) were performed. Predictive 3D-QSAR models were obtained with $q^2$ values of 0.869 and 0.872 and $r^2_{ncv}$ values of 0.983 and 0.993 for CoMFA and CoMSIA, respectively. These results demonstrate that CoMFA and CoMSIA models could be reliably used in the design of novel cytotoxic agents.

Effect of Bioactive Glass Addition to the TTCP/DCPA Based Injectable Bone Substitute for Improved Biocompatibility

  • Sadiasa, Alexander;Sarkar, Swapan Kumar;Franco, Rose Ann;Yang, Hun-Mo;Lee, Byong-Taek
    • Proceedings of the Materials Research Society of Korea Conference
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    • 2011.05a
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    • pp.52.1-52.1
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    • 2011
  • In this work, the effect of the addition of bioactive glass in the biocompatibility and mechanical behavior of conventional TTCP/DCPA based bone cement were investigated. The cement was initially modified with chitosan and HPMC which cross-linked with citric acid to improved mechanical properties.The injectable bone substitutes were further modified by adding varying amounts of bioactive glass (0%, 10%, 20% and 30%) and its effects on the biocompatibility of the material were studied. Afterbio-glass powders were mixed with the optimized composition for HPMC and citric acid content,the IBS was incubated at $37^{\circ}C$ at different time intervals and showed progressive formation of HAp with increasing time. Mechanical properties like Vickers hardness and compressive strength were found to increase with the increasing amount of bioactive glass addition and that setting time was shortened. The fabricated IBS morphologies were further characterized using SEM. MTT assay was performed to check the cell cytotoxicity and cell proliferation for 1, 3 and 5 days. Cell morphology, adhesion and proliferation behavior of cell in the IBS by culturing MG-63 cells on the IBS for 20, 60 and 90 mins and 1, 3 and 5 days was also investigated. All the results showed increasing biocompatibility as the bioglass content increased. MTT results found the materials to be cytocompatible and SEM images showed that cells attached and proliferated successfully.

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Cytotoxicity and Antimicrobial Effects of Extracts from Salvia miltiorrhiza (단삼 추출물의 세포독성과 항균효과)

  • Kwag, Jung-Sook;Baek, Seung-Hwa
    • Korean Journal of Pharmacognosy
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    • v.34 no.4 s.135
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    • pp.293-296
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    • 2003
  • This study was carried out to evaluate cytotoxic effects of Salvia miltiorrhiza extracts on NIH 3T3 fibroblasts and KB cell lines. Disruptions in cell organelles were determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The comparison of $IC_{50}$ of values of Salvia miltiorrhiza extracts in KB cell lines showed that their susceptibility to these extracts decreased in the following order: hexane extract > chloroform extract > methanol extract> dichloromethane extract > ethyl acetate extract>ethanol extract by the MTT method. The dried roots of Salvia miltiorrhiza was extracted several solvents, and then antimicrobial activity was investigated. The minimal inhibitory concentrations (MIC's) of the extract against microorganisms were also examined. Amtimicrobial activity of ketoconazol as reference was compared to those of extracts of hexane, chloroform, dichloromethane, ethyl acetate, ethanol and methanol. The antimicrobial activity of all extracts from the sample had growth inhibition activity against gram-negative bacteria, gram-positive bacteria and fungi. These results suggest that the hexane and chloroform soluble extracts of Salvia miltiorrhiza may be a valuable choice for the studies on the tumor cell lines and growth inhibition activity.

Cytotoxic Saponins from the Root of Dipsacus asper Wall

  • Hung Tran Manh;Jin WenYi;Thuong Phuong Thien;Song Kyung Sik;Seong Yeon Hee;Bae KiHwan
    • Archives of Pharmacal Research
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    • v.28 no.9
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    • pp.1053-1056
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    • 2005
  • Cytotoxic activitiy of seven hederagenin saponins isolated from the root of Dipsacus asper were investigated in vitro against L1210, HL-60 and SK-OV-3 tumor cell lines by the MTT method. $3-O-\alpha-L-rhamnopyranosyl-(1{\rightarrow}2)-\alpha-L -arabinopyranosyl$ hederagenin (2),\;$3-O-\beta-D­xylopyranosyl-( 1{\rightarrow}3)-\alpha-L-Rhamnopyranosyl-(1{\rightarrow}2)-\alpha-L -arabinopyranosyl$ hederagenin (6) and $3-O-\beta-D-glucopyranosyl-(1{\rightarrow}3)-\alpha-L-rhamnopyranosyl-( 1{\rightarrow}2)-\alpha-L-arabinopyranosyl$ hederagenin (7) exhibited the potent cytotoxicity against the three tumor cell lines with $IC_{50}$ values ranging from 4.7 to 8.7 ${\mu}g/mL$, with the exception of compound 7, which exhibited weak cytotoxic activity against SK-OV-3 $(IC_{50}\;22.5\;{\mu}g/mL)$. Other compounds did not exhibit any cytotoxic activity $(IC_{50}>30{\mu}g/mL)$.

Effects on the Apoptosis of U937 cell line by n-butanol Extracts of Drynariae Rhizoma (골쇄보(骨碎補) 부탄올 추출물의 U937세포주 세포자멸사에 미치는 효과)

  • Cho, Sung-Yeon;Jeong, Han-Sol
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.20 no.2
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    • pp.358-364
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    • 2006
  • To investigate the possible mechanism of Drynariae Rhizoma extracts as a candidate of anti-cancer drug, I examined the effects of Drynariae Rhizoma extracts on the apoptosis of U937 cell line. MTT assay, flow cytometric analysis, SDS-polyacrylamide gel electrophoresis, Western blot analysis, and RT-PCR were performed. Drynariae Rhizoma extracts treatment reduced the cell viablilty of U937 cells in a dose-dependent manner, which was associated with induction of apoptotic cell death. Drynariae Rhizoma extracts treatment also reduced the levels of Bcl-xL anti-apoptotic protein expression and increased the levels of caspase-3, p53, pro-apoptotic protein, in U937 cells. RT-PCR data revealed that the level of bcl-2, bcl-xL mRNA expressions decreased in a dose-dependent manner. These findings suggest that Drynariae Rhizoma extracts may have induction of apoptotic cell death via regulation of several growth regulatory gene products. The abbreviations used are: FBS, fetal bovine serum; PBS, phosphate buffered saline; PI, propidium iodide; OD, optical density; DiOC6, 3,3-dihexyloxa carbcyanine iodide; MTT, 3 [4-5-dimethylthiazol-2-yl] -2-diphenyltetrazolium bromide.

In vitro Screening of Seaweed Extract on the Proliferation of Mouse Spleen and Thymus Cell

  • Seo, Young-Wan;Kang, Sung-Ho;Lee, Hee-Jung;Kim, You-Ah;Youn, Hyun-Joo;Lee, Burm-Jong;Chung, Ho-Sung
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.2
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    • pp.160-163
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    • 2006
  • A total number of 31 types of seaweed were assessed with regard to their effects on the proliferation of mouse spleen and thymus cells in a culture, using an MTT reduction assay. Acetone:dichloromethane (1:1) extracts of three seaweed plants: Derbesia marina, Sargassum sp., and Hisikia fuziformis, exhibited significantly positive effects on the survival of mouse spleen and thymus cells in vitro. The acetone:dichloromethane (1:1) extracts of Sargassum sp., in particular, much more potent effects on thymus cell activation than did any of the other types of seaweed. However, the methanol extracts of Sargassum ringgoldianium and Chondrus crispus exerted a stimulatory influence only on the proliferation of mouse spleen cells, whereas the methanol extracts of Grateloupia lanceolata exhibited significant cell proliferation properties in both spleen and thymus cells.

Ferutinin, an Apoptosis Inducing Terpenoid from Ferula ovina

  • Matin, Maryam Moghaddam;Nakhaeizadeh, Hossein;Bahrami, Ahamd Reza;Iranshahi, Mehrdad;Arghiani, Nahid;Rassouli, Fatemeh Behnam
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.5
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    • pp.2123-2128
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    • 2014
  • A current hurdle in cancer management is the intrinsic or acquired resistance of cancer cells to chemical agents that restricts the efficacy of therapeutic strategies. Accordingly, there is an increasing desire to discover new natural compounds with selective toxicity to combat malignancies. In present study, the cytotoxic and apoptosis-inducing activities of ferutinin, a terpenoid derivative from Ferula ovina, were investigated on human breast (MCF7) and bladder (TCC) cancer cells as well as normal fibroblasts (HFF3).The toxicity and DNA damage inducing effects of ferutinin were studied by MTT and comet assays, DAPI and PI staining and DNA laddering. The $IC_{50}$ values of ferutinin were identified and compared with routine prescribed drugs, doxorubicin and vincristine, by MTT test. Alkaline comet assay and DAPI staining revealed DNA damage due to ferutinin, which was significantly (p<0.001) higher in MCF7 and TCC than HFF3 cells. Apoptosis induction was evidenced by PI staining and DNA laddering. Our results suggest that ferutinin could be considered as an effective anticancer agent for future in vivo and clinical experiments.