• Title/Summary/Keyword: MTS Assay

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Study on the Anti-Cancer Effect of Acanthopanax Sessiliflorus (오가피(五加皮)의 항암효과에 관한 연구)

  • Kim, Young-Chul;Woo, Hong-Jung;Lee, Jang-Hoon
    • The Journal of Korean Medicine
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    • v.20 no.3 s.39
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    • pp.54-65
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    • 1999
  • Objectives: Hepatoma is a very serious disease in Korea and vvorldwiclc. Hepatitis B vims (HBV) has proved the most significant cause of hepatoma. We canied out this study to investigate the effect of Acanthopanax sessilifloms on inhibiting cell proliferation and DNA synthesis in HepG2.2.15 cell line and on inhibiting phosphorilation of oncogene (MAP kinase) in NIT/3T3-HBx ceIl. Methods: To investigate the anti-cancer effect of Acanthopanax sessiliflorus, we did the CellTiter 96 Aqueous Non-radioactive Cell Proliferation assay (Promega); MTS/PMS assay, [$^3H$]-thymicline incorporation assay, and we measured the gene expression through westem blotting. Results: Acanthopanax sessiliflorus showed an inhibiting effect on the increase of HepG2.2.15 in the NTS/PMS assay. It also showed an inhibiting effect on DNA synthesis of HepG2.2.15 in the [$^3H$]-thymidine incorporation assay. Acanthopanax sessiliflorus showed an inhibiting effect of phosphorilation of MAP kinase in HBV - X genes. too. Conclusions: The results suggested that this herb had an anti cancer effect. We may discover an effective anti-cancer herb medicine through further studies on this herb medicine.

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The Change of Cell-cycle Related Proteins and Tumor Suppressive Effect in Non-small Cell Lung Cancer Cell Line after Transfection of p16(MTS1) Gene (폐암세포에 p16 (MTS1) 유전자 주입후 암생성능의 변화 및 세포주기관련 단백질의 변동에 관한 연구)

  • Kim, Young-Whan;Kim, Jae-Yeol;Yoo, Chul-Gyu;Han, Sung-Koo;Shim, Young-Soo;Lee, Kye-Young
    • Tuberculosis and Respiratory Diseases
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    • v.44 no.4
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    • pp.796-805
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    • 1997
  • Background : It is clear that deregulation of cell cycle progression is a hallmark of neoplastic transformation and genes involved in the $G_1$/S transition of the cell cycle are especially frequent targets for mutations in human cancers, including lung cancer. p16 gene product, one of the G1 cell-cycle related proteins, that is recently identified plays an important role in the negative regulation of the the kinase activity of the cyclin dependent kinase (cdk) enzymes. Therefore p16 gene is known to be an important tumor suppressor gene and is also called MTS1 (multiple tumor suppressor 1). No more oncogenes have been reported to be frequently related to multiple different malignancies than the alterations of p16 gene. Especially when it comes to non-small cell lung cancer, there was no expression of p16 in more than 70% of cell lines examined. And also it is speculated that p16 gene could exert a key role in the development of non-small cell lung cancer. This study was designed to evaluate whether p16 gene could be used as a candidate for gene therapy of non-small cell lung cancer. Methods : After the extraction of total RNA from normal fibroblast cell line and subsequent reverse transcriptase reaction and polymerase chain reaction, the amplified p16 cDNA was subcloned into eukaryotic expression plasmid vector, pRC-CMV. The constructed pRC-CMV-p16 was transfected into the NCI-H441 NSCLC cell line using lipofectin. The changes of G1 cell-cycle related proteins were investigated with Western blot analysis and immunoprecipitation after extraction of proteins from cell lysates and tumor suppressive effect was observed by clonogenic assay. Results : (1) p16(-) NCI-H441 cell line transfected with pRC-CMV-p16 showed the formation of p16 : cdk 4 complex and decreased phosphorylated Rb protein, while control cell line did not. (2) Clonogenic assay demonstrated that the number of colony formation was markedly decreased in p16(-) NCI-H441 cell line transfected with pRC-CMV-p16 than the control cell line. Conclusion : It is confirmed that the expression of p16 protein in p16 absent NSCLC cell line with the gene transfection leads to p16 : cdk4 complex formation, subsequent decrease of phosphorylated pRb protein and ultimately tumor suppressive effects. And also it provides the foundation for the application of p16 gene as a important candidate for the gene therapy of NSCLC.

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Antitumor activity and 3D-Histoculture drug response assay of Novel trans- ditrifluoroacetato,malonato-1,4-butanediamine Pt(IV) complex, K104

  • Kwon, Young-Ee;Kim, Kuk-Hwan;Oh, Bong-Un;Kim, Kap-Joon
    • Proceedings of the PSK Conference
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    • 2003.04a
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    • pp.132.1-132.1
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    • 2003
  • Novel trans-ditrifluoroacetato, malonato-1, 4-butanediamine Pt(IV) complex, K104 was synthesized as a chemotherpeutic. The cytotoxicity of K104 against various human cancer cell lines were evaluated by MTS assay in vitro. The IC50 values of K104 ranged 15.83-25.83 $\mu\textrm{M}$, compared to CBCDA ranged 23.24-69.6 $\mu\textrm{M}$. Among several cancer cell lines, K104 showed more potent than CBCDA in colon cancer cell lines. (omitted)

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The Effect of Bojungykgitang-Chunbang on Activity of CD4+ T cell

  • Lee Tae Hyong;Kang Hee;Myung Eu Gene;Shim Bum Sang;Choi Seung Hoon;Kim Sung Hun;Ahn Kyoo Seok
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.18 no.2
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    • pp.580-585
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    • 2004
  • BJYGC is often clinically used as a treatment of allergic rhinitis. This study was aimed to find out the effect BJYGC would have on the helper T cell, and how it can promote the subsets of helper T cells to regain their balance that they lost due to immunological diseases. Splenocytes were prepared from BALB/c mice was cultured without stimulation in the presence of BJYGC for 48 hr. The viability of CD4 T cells from Balb/c mouse were measured at various concentrations of BJYGC using the MTS assay. It was somewhat increased up to concentration of 400 ㎍/ml, but did not show any significant difference. Proliferation was measured using the MTS assay, CD4 Th cells were stimulated with anti-CD3/28 in the presence of BJYGC for 48 hr. As evidence for rapid T cell activation, CD25 expression by flow cytometry was evaluated at 10, 50, 100 and 200 ㎍/㎖ of BJYGC. Th cell differentiation experiments were performed to examine whether BJYGC can affect the Th polarization process. CD4 T cells were activated in culture under neutral, Th1-polarized or Th2-polarized conditions in the presence of BJYGC at 10, 100 and 200 ㎍/㎖. Cytokine production was measured by ELISA. This experiment proved that BJYGC could inhibit the secretion of both IL-4 and IFN-γ in neutral condition and polarized condition, too. Considering that BJYGC shows an excellent effect on treating allergies, the author can conclude that its pharmacological action may be associated with decreased IL-4 and, it may also regulate IFN-γ depending the host's need. Also, it was discovered that Th1 cell was pathologic in chronic inflammatory tissue specific diseases, such as insulin dependent diabetes mellitus, multiple sclerosis, RA, and uveitis. We are counting on the BJYGC to be able to control the tendency of Th1 cell predominancy in an immune reaction.

Antioxidant Effect of Traditional food Ingredient (전통 식재료의 항산화 효과)

  • Lee, Jae-Hyeok;Park, Jeong-Sook
    • Journal of the Korea Convergence Society
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    • v.11 no.2
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    • pp.309-314
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    • 2020
  • This study was designed to investigate the antioxidant effects of 10 kinds of medicinal plants and vegetable extracts and total extracts. The cytotoxicity was measured by MTS assay, and the antioxidant activity was measured by DPPH free radical scavenging activity and Riboflavin-derived superoxide inhibitory activity (SQA). As a result, cytotoxicity was safe for all 10 medicinal plants, vegetable extracts and total extracts. DPPH free radical scavenging ability was observed in Cinnamomum cassia Blume, Eugenia caryophyllata Thunb. Arctium lappa, Total extract was excellent, and Riboflavin-derived superoxide inhibitory activity (SQA) was found in Cinnamomum cassia Blume, Arctium lappa, Prunus mume Sieb. et Zucc., Excellent, but total extract showed the best antioxidant effect. As a result of comparing the antioxidant effects of medicinal plants and vegetables using traditional ingredients, the antioxidant activity was increased when used as a mixture than when used alone. It is considered that it can be used as an antioxidant functional material, and it is expected to be of value when developing antioxidant material in the future.

Investigation of the Effect of Water Extract of Lithospermi Radix on the Expression of IL-1β, TNF-α and iNOS Genes in Raw 264.7 Cells (자초(紫草) 열수 추출물의 RAW 264.7 세포에서 IL-1β, TNF-α, iNOS 유전자 발현에 미치는 영향 연구)

  • Cho, Nam Joon;Choi, Young Ho;Lee, Woong Hee;Kim, Kee Kwang;Han, Hyo Sang
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.31 no.4
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    • pp.220-225
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    • 2017
  • Lithospermi Radix (LR) is known to have an anti-inflammatory effect. However, the mechanisms are not well known. In this study, LPS-induced mouse RAW 264.7 macrophage cells were treated with LR to investigate the time-dependent inflammation response of LR. RAW 264.7 cells were treated with various concentrations of LR for 24 hours, followed by MTS assay. Cell viability was increased at all experimental concentrations. The mRNA expression levels of $IL-1{\beta}$, $TNF-{\alpha}$ and iNOS were increased by treatment of RAW 264.7 cells with LR at a concentration of $200{\mu}g/ml$ for 6 hours and 24 hours. Treatment of LR with $200{\mu}g/ml$ concentration for 6 hours promoted mRNA expression levels of $IL-1{\beta}$, $TNF-{\alpha}$ and iNOS in LPS-induced RAW 264.7 cells. However, $IL-1{\beta}$, $TNF-{\alpha}$ and iNOS mRNA expression was suppressed by treatment of LR with $200{\mu}g/ml$ concentration for 24 hours in LPS-induced RAW 264.7 cells. These results suggest that the effect on inflammation of LR is promptly promoted and then to rapidly alleviate the inflammatory reaction. This study proposes that the time-dependent activities of herbal medicine is a very important factor in analyzing the anti-inflammatory effect of various herbal medicines including LR.

Effect of Brussels Sprouts Extract on Inflammatory Cytokine Inhibition (방울양배추 추출물의 염증성 사이토카인 억제에 미치는 영향)

  • Jae-Hyeok Lee;Jeong-Sook Park
    • Journal of Industrial Convergence
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    • v.21 no.8
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    • pp.69-74
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    • 2023
  • This paper was conducted to examine the effect of Brussels Sprouts Extract on the inhibition of pro-inflammatory cytokines. The inflammatory response is manifested by mediators such as reactive oxygen species and inflammatory cytokines such as TNF-α, IL-1β, and IL-8. Therefore, this paper examined the toxicity to cells using the MTS assay, stimulated RAW264.7 macrophages with lipopolysaccharide (LPS), and stimulated reactive oxygen species such as NO and TNF-α, IL-1β, and IL-8. Inhibition of inflammatory cytokines after treatment with 10 mg/mL, 100 mg/mL, and 1000 mg/mL of Brussels Sprouts Extract was investigated. As a result of the experiment, Brussels Sprouts Extract inhibited NO production, TNF-α and IL-8 in a concentration-dependent manner without cytotoxicity, and showed significant inhibition especially at a concentration of 1000 mg/mL. Brussels Sprouts Extract, which inhibits the production of inflammatory cytokines, suggests the possibility of reducing inflammatory response and controlling inflammation, and can be seen as providing potential as a health functional food or prevention and treatment of inflammation.

Identification of the Antioxidative Function of Metallothionein by Oxidation of NADPH and Production of Nitrite (NADPH의 산화반응과 아질산 생성반응에 의한 Metallothionein 의 항산화적 기능 확인)

  • Kim Kwan-Chun;Kim Joon-Tae;Kim Hee-Joung
    • Journal of environmental and Sanitary engineering
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    • v.21 no.1 s.59
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    • pp.52-57
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    • 2006
  • Metallothioneins(MTs) belong to the class of low molecular weight proteins. Recently, it has been suggested that MTs may playa direct role in cellular defense against oxidative stress by functioning as antioxidants. Oxidative damage to different cellular components makes a major contribution to many pathogenenesses. Several studies have demonstrated that MT is able to quench a wide range of reactive oxygen species at a higher efficiency than other well known antioxidants such as superoxide dismutate(SOD). The present study was designed to evaluate the effect of MT on the activities of the reactive oxygen species removal system. MT showed the scavenging of superoxide in the SOD assay system in the presence or absence of SOD. When MT was added to nicotinamide adenine dinucleotide phosphate(NADPH) oxidation system in presence of fixed amount of SOD increase the breakdown rate of superoxide. When MT was added to the system that form nitrite from hydroxylammonium chloride, the formation of nitrite was inhibit. We concluded that the function of MT as antioxidant might have an effect on the level of superoxide scavenging.

Orostachys japonicus DW and EtOH Extracts Induce Apoptosis in Cholangiocarcinoma Cell Line SNU-1079

  • Choi, Eun Sol;Lee, Jang Hoon
    • The Journal of Korean Medicine
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    • v.36 no.4
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    • pp.19-34
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    • 2015
  • Objectives: This study was performed to investigate the anti-tumor effect of O. japonicus extracts on intrahepatic cholangiocarcinoma cell line SNU-1079. Methods: Cholangiocarcinoma SNU-1079 cells were treated with various concentrations of O. japonicus DW and EtOH extracts ($0-300{\mu}g/ml$) for 24, 48 or 72 h. Cell viability was evaluated through a PMS/MTS assay, and the apoptosis rate was examined through ELISA assay and flow cytometry analysis. The mRNA expression of apoptosis- and cell cycle progression-related genes (Bcl-2, Mcl-1, Bax, Survivin, Cyclin D1, and p21) was evaluated using real-time PCR, and the caspase activity was examined using immunoblot analysis. Results: O. japonicus extracts inhibited cell proliferation and increased apoptosis rate in both ELISA assay and flow cytometry analysis. O. japonicus extracts decreased Bcl-2, Mcl-1, Survivin, and Cyclin D1 mRNA expression and increased Bax mRNA level. O. japonicus extracts also increased Caspase-3 activation. Overall, O. japonicus DW extracts were more effective than EtOH extracts. Conclusions: O. japonicus inhibited cell proliferation and induced apoptosis in SNU-1079 cells via mitochondria -mediated intrinsic pathway, which leads to Caspase-3 activation. The results indicate that O. japonicus is a potential therapeutic herb with anti-tumor effect against intrahepatic cholangiocarcinoma.

Effect of Anti-oxidant, Anti-inflammatory and Anti-invasive of PMA-induced Matrix Metalloproteinase (MMP-2) and MMP-9 Activities of Water Extract and Solvent Fractions of Saururus Chinensis (삼백초 물 추출물과 유기용매 분획물의 항산화, 항염증 및 PMA에 의해 유도된 MMP-2 및 MMP-9활성 침윤 억제 효과)

  • Kim, Jun-Ho;Kim, Eun-Jung
    • Journal of Life Science
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    • v.26 no.5
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    • pp.584-591
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    • 2016
  • Saururus chinensis is a perennial plants, its flavonoid compound is known to exhibit anti-oxidative activity. This study was aimed to investigate the effect of Water Extract and Solvent Fractions of Saururus chinensis on antioxidant, anti-inflammatory and anti-invasive of Phorbol 12-myristate 13-acetate (PMA)-induced matrix metalloproteinase (MMP-2) and MMP-9 activities. Plant samples were fractionated into hexane, CHCl3, ethyl acetate, butanol, and water fractions, and each of these was assayed individually. The water fraction showed the highest extraction yield at 9.25%(w/w). Anti-oxidative activity was analyzed by DPPH assay. Cell viability was detected by the MTS assay. Anti-inflammatory activity was assayed by the nitric oxide (NO) production in mouse macrophage Raw 264.7 cells. The activity and mRNA expression of MMP-2 and MMP-9 in human oral squamous carcinoma YD-10B cells were examined by zymography and RT-PCR. As results, MMP-2/-9 activation was increased in PMA induced YD-10B cells. In PMA-treated YD-10B cells, the increased mRNA expression and protein activation of MMP-2/-9 were significantly inhibited in the ethyl acetate fraction. The ethyl acetate fraction showed the highest anti-oxidative activity at 73.38%. The ethyl acetate fraction at non-cytotoxic concentrations significantly exhibited the anti-inflammatory activity of Raw 264.7 cells in dose-dependent manner. In conclusion, these findings demonstrate that the ethyl acetate fraction obtained from a chinensis water extract potentiates a promising therapeutic anti-invasive agent and, therefore, as an anti-cancer drug for cancer prevention and therapy in oral cancer.