• 제목/요약/키워드: IC-RT-PCR

검색결과 71건 처리시간 0.038초

Differential inhibitory effects of alpha-viniferin, resveratrol trimer on inflammatory mediators

  • Chung, Eun-Yong;Min, Kyung-Rak;Kim, Young-Soo
    • 대한약학회:학술대회논문집
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    • 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.1
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    • pp.213.1-213.1
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    • 2003
  • Alpha-viniferin was isolated from Carex humilis (Cyperaceae), and showed anti-inflammatory effects on carrageenin or histamine-induced paw edema in mice. To understand mode of the anti-inflammatory action. effects of alpha-viniferin on cyclooxygenase (COX)-2, iNOS, oxygen radicals and proinflammatory cytokines have been analyzed. Alpha-viniferin showed selective inhibitory effect with an IC50 value of 5 $\mu\textrm{m}$ on COX-2 activity but showed weak inhibitory effect on the synthesis of COX-2 transcript which was identified by RT-PCR. (omitted)

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Aqueous Extract of Anticancer Drug CRUEL Herbomineral Formulation Capsules Exerts Anti-proliferative Effects in Renal Cell Carcinoma Cell Lines

  • Verma, Shiv Prakash;Sisoudiya, Saumya;Das, Parimal
    • Asian Pacific Journal of Cancer Prevention
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    • 제16권18호
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    • pp.8419-8423
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    • 2016
  • Purpose: Anti-cancer activity evaluation of aqueous extract of CRUEL (herbomineral formulation) capsules on renal cell carcinoma cell lines, and exploration of mechanisms of cell death. Materials and Methods: To detect the cytotoxic dose concentration in renal cell carcinoma (RCC) cells, MTT assays were performed and morphological changes after treatment were observed by inverted microscopy. Drug effects against RCC cell lines were assessed with reference to cell cycle distribution (flow cytometry), anti-metastatic potential (wound healing assay) and autophagy(RT-PCR). Results: CRUEL showed anti-proliferative effects against RCC tumor cell lines with an IC50 value of ${\approx}4mg/mL$ in vitro., while inducing cell cycle arrest at S-phase of cell cycle and inhibiting wound healing. LC3 was found to be up-regulated after drug treatment in RT-PCR resulting in an autophagy mode of cell death. Conclusions: This study provides the experimental validation for antitumor activity of CRUEL.

BmKn-2 Scorpion Venom Peptide for Killing Oral Cancer Cells by Apoptosis

  • Tong-ngam, Pirut;Roytrakul, Sittiruk;Sritanaudomchai, Hathaitip
    • Asian Pacific Journal of Cancer Prevention
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    • 제16권7호
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    • pp.2807-2811
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    • 2015
  • Scorpion venom peptides recently have attracted attention as alternative chemotherapeutic agents that may overcome the limitations of current drugs, providing specific cytotoxicity for cancer cells with an ability to bypass multidrug-resistance mechanisms, additive effects in combination therapy and safety. In the present study, BmKn-2 scorpion venom peptide and its derivatives were chosen for assessment of anticancer activities. BmKn-2 was identified as the most effective against human oral squamous cells carcinoma cell line (HSC-4) by screening assays with an $IC_{50}$ value of $29{\mu}g/ml$. The BmKn-2 peptide killed HSC-4 cells through induction of apoptosis, as confirmed by phase contrast microscopy and RT-PCR techniques. Typical morphological features of apoptosis including cell shrinkage and rounding characteristics were observed in treated HSC-4 cells. The results were further confirmed by increased expression of pro-apoptotic genes such as caspase-3, -7, and -9 but decrease mRNA level of anti-apoptotic BCL-2 in BmKn-2 treated cells, as determined by RT-PCR assay. In summary, the BmKn-2 scorpion venom peptide demonstrates specific membrane binding, growth inhibition and apoptogenic activity against human oral cancer cells.

Diosgenin Inhibits hTERT Gene Expression in the A549 Lung Cancer Cell Line

  • Mohammad, Rahmati Yamchi;Somayyeh, Ghareghomi;Gholamreza, Haddadchi;Majid, Mobasseri;Yousef, Rasmi
    • Asian Pacific Journal of Cancer Prevention
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    • 제14권11호
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    • pp.6945-6948
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    • 2013
  • Background: Diosgenin, a steroidal saponin from a therapeutic herb, fenugreek (Trigonellafoenum-graceum L.), has been recognized to have anticancer properties. Telomerase activity is not detected in typical healthy cells, while in cancer cell telomerase expression is reactivated, therefore providing a promising cancer therapeutic target. Materials and Methods: We studied the inhibitory effect of diosgenin on human telomerase reverse transcriptase gene (hTERT) expression which is critical for telomerase activity. MTT- assays and qRT-PCR analysis were conducted to assess cytotoxicity and hTERT gene expression inhibition effects, respectively. Results: MTT results showed that $IC_{50}$ values for 24, 48 and 72h after treatment were 47, 44 and $43{\mu}M$, respectively. Culturing cells with diosgenin treatment caused down-regulation of hTERT expression. Discussion: These results show that diosgenin inhibits telomerase activity by down-regulation of hTERT gene expression in the A549 lung cancer cell line.

Update on Distribution and Genetic Variability of Plum pox virus Strains in Bulgaria

  • Kamenova, Ivanka;Borisova, Anelija
    • The Plant Pathology Journal
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    • 제35권3호
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    • pp.243-256
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    • 2019
  • Field surveys for Plum pox virus (PPV) infection were conducted in stone fruit orchards all over Bulgaria. In total, 1168 out of 3020 leaf samples from cultivated Prunus spp. and wildly growing P. cerasifera trees reacted positive for PPV in DASI-ELISA with the universal monoclonal antibody (MAb) 5B. Further ELISA analyses showed that 987 and 127 isolates belonged to PPV-M and PPV-D serotypes, respectively. The plum and P. cerasifera showed 82.0% and 50.5% levels of infection, respectively followed by the peach (40.0%) and the apricot (32.0%). Five hundred fifty one PPV isolates were further typed by IC-RT-PCR with PPV-Rec, -M and -D-specific primers, targeting (Cter)NIb-(Nter) CP genome region, as 125 isolates were sequenced. The results revealed the presence of PPV-Rec, PPV-M and PPV-D and mixed infections of these strains. PPV-Rec was the most prevalent strain (49.0%), followed by PPV-M (40.1%), while PPV-D was the less spread strain (8.2%). PPV-Rec was the most common strain in plums, including the eight "old-aged" trees from the region of the first Sharka discovery. PPV-M was the most prevalent strain in peach and apricot. Phylogenetic analyses on (Cter)NIb-(Nter)CP of the isolates were performed. PPV-Rec isolates formed a homogeneous group, while PPV-M isolates split into PPV-Ma and PPV-Mb subgroups. Five separated clades were formed by the analyzed PPV-D isolates. Nucleotide sequences of the partial CP coding region of the analyzed isolates revealed a slightly higher intra-strain genetic variability in PPV-Rec and PPV-M isolates, while that of PPV-D strain isolates was higher from the reported for these strains.

Aromatic diamine JSH-21 inhibits LPS-induced NO production by targeting NF-kB signaling

  • Shin, Hyun-Mo;Jung, Sang-Hun;Min, Kyung-Rak;Kim, Young-Soo
    • 대한약학회:학술대회논문집
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    • 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2-2
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    • pp.151.3-152
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    • 2003
  • Aromatic diamine JSH-21 showed an IC50 value of 9.2 uM with 74.5% inhibition at 30 uM, 53.5% at 10 uM and 24.5% at 3 uM on LPS-induced NO production in murine macrophages Raw 264.7. To examine whether inhibitory effect on NO production by JSH-21 was attributed to influence on iNOS expression, iNOS transcript and protein were analyzed by sequantitative RT-PCR and immunoblot analysis. Consistent with previous result on NO production, treatment of the Raw 264.7 cells with JSH-21 decreased the LPS-induced expression of iNOS transcript and protein in a dose-dependent manner with IC50 values of about 10 uM. (omitted)

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Sophoricoside analogs inhibit COX isozymes but not iNOS and TNF in LPS-stimulated macrophages Raw264.7

  • Kim, Byung-Hak;Min, Kyung-Rak;Kim, Young-Soo
    • 대한약학회:학술대회논문집
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    • 대한약학회 2002년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2
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    • pp.320.2-320.2
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    • 2002
  • Macrophages activated by lipopolysaccharide (LPS) are known to induce several proinflammatory proteins including COX-2. iNOS and TNF which produce chemical mediators involved in inflammatory response. Sophoricoside and its analogs (genistin, genistein and orobol) from Sophora japonica (Leguminosae) showed differential inhibitory effects on COX-1 and 2 activities. Sophoricoside and genistin shwoed IC50 values of 4 uM and 6 uM on COX-2 activity and of 1,497 uM and 135 uM on COX-1 activity, respectively. Genistein and orobol showed IC50 values of 3 uM on COX-2 activity and of 28 uM and 18 uM on COX-1 activity. respectively. Therefore. the legume isoflavonoids to be selective COX-2 inhibitors. However. sophoricoside and its analogs did not show inhibitory effects of COX-2, iNos and TNF transcripts. which were identified by the RT-PCR.

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Antioxidant and Antiproliferative Activities of Methanolic Extract from Celandine

  • Hu, Weicheng;Wang, Myeong-Hyeon
    • Food Science and Biotechnology
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    • 제18권1호
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    • pp.207-212
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    • 2009
  • Celandine (Chelidonium majus, family Papaveraceae) is an herb used extensively in traditional Korean medicine. To investigate its antioxidant and antiproliferative activities, the methanolic extract of celandine was introduced. The antioxidant properties of the extract were tested using various in vitro systems, including hydroxyl radical scavenging assay, DNA damage protection assay, 1,1-diphenyll-2-2-pricylhydrazyl (DPPH) free radical scavenging activity, metal chelating activity, and reducing power assay. The extract exhibited stronger antioxidant activity ($IC_{50}=7.92{\mu}g/mL$) against hydroxyl radicals in the Fenton system than butylated hydroxyanisole ($IC_{50}=51.46{\mu}g/mL$) and $\alpha$-tocopherol ($IC_{50}=67.48{\mu}g/mL$). Likewise, damage to the plasmid pBR 322 induced by hydroxyl radicals was found to be protected by the extract at a concentration of $400{\mu}g/mL$. Cellular proliferation and the induction of apoptosis were also examined by a cellular proliferation assay, flow cytometry, and mRNA expression analysis. Taken together, the extract significantly inhibited the growth of HT-29 cells in a concentration- and time-dependent manner, and gradually increased both the proportion of apoptotic cells and the expression of caspase-3. Overall, our research suggests that celandine possesses antioxidant and antiproliferative properties.

Inhibition of Tumor Growth in Vitro by a Combination of Extracts from Rosa Roxburghii Tratt and Fagopyrum Cymosum

  • Liu, Wei;Li, Su-Yi;Huang, Xin-En;Cui, Jiu-Jie;Zhao, Ting;Zhang, Hua
    • Asian Pacific Journal of Cancer Prevention
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    • 제13권5호
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    • pp.2409-2414
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    • 2012
  • Objective: Traditional Chinese herbal medicines have a very long history. Rosa roxburghii Tratt and Fagopyrum cymosum are two examples of plants which are reputed to have benefits in improving immune responses, enhancing digestive ability and demonstrating anti-aging effects. Some evidence indicates that herbal medicine soups containing extracts from the two in combination have efficacy in treating malignant tumors. However, the underlying mechanisms are far from well understood. The present study was therefore undertaken to evaluate anticancer effects and explore molecular mechanisms in vitro. Methods: Proliferation and apoptosis were assessed with three carcinoma cell lines (human esophageal squamous carcinoma CaEs-17, human gastric carcinoma SGC-7901 and pulmonary carcinoma A549) by MTT assay and flow cytometry, respectively, after exposure to extract from Rosa roxburghii Tratt (CL) and extract from Fagopyrum cymosum (FR). $IC_{30}$ of CL and FR were obtained by MTT assay. Tumor cells were divided into four groups : control with no exposure to CL or FR; CL with $IC_{30}$ CL; FR with $IC_{30}$ FR; CL+FR group with 1/2 ($IC_{30}$ CL + $IC_{30}$ FR). RT-PCR and Western blot analysis were used to detect the expression of Ki-67, Bax and Bcl-2 at mRNA and protein levels. Results: Compared with the CL or FR groups, the combination of CL+FR showed significant inhibition of cell growth and increase in apoptosis; the mRNA and protein expression levels of Ki-67 and Bcl-2 in CL+FR group were all greatly decreased, while the expression of Bax was markedly increased. Conclusions: These results indicate that the synergistic antitumor effects of combination of CL and FR are related to inhibition of proliferation and induction of apoptosis.

자외선이 조사된 사람 피부 섬유아세포에서 권백의 항산화와 MMP 발현에 미치는 영향에 대한 연구 (Antioxidation and Inhibition of Matrix Metalloproteinase in UV-irradiated Human Dermal Fibroblast by Selaginella tamariscina)

  • 심관섭;김진화;김진희;이범천;표형배
    • 대한화장품학회지
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    • 제32권2호
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    • pp.75-79
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    • 2006
  • 다양한 약용식물의 피부노화에 미치는 영향을 평가하였으며, 이 중 권백(Selaginella tamariscina)은 동양에서 암환자 치료를 위한 전통약용식물로 알려져 있다. 우리는 피부노화에 대한 화장품 소재로 권백에 대한 다양한 생물학적 평가를 하였다. 권백의 항산화 효과를 알아보기 위해 DPPH radical과 superoxide anion radical 소거효과를 측정하였다. 그 결과 DPPH radical의 $IC_{50}$ 값은 $65.1{\mu}g/mL$이고, xanthine/xanthine oxidase에 의한 superoxide anion radical의 $IC_{50}$ 값은 $40.9 {\mu}g/mL$이었다. 세포내 활성산소 소거평가를 위해 사람 진피 섬유아세포(human dermal fibroblast)를 배양하여 UVB $20 mJ/cm^2$에 의해 증가된 세포내 활성산소(ROS)가 권백을 처리함으로써 활성산소 소거효과가 증가하였다. 사람 진피 섬유아세포에서 UVA에 의해 발현된 MMP-1 단백질과 mRNA가 권백에 의해 농도 의존적으로 감소하였다. 뿐만 아니라, 권백은 zymography와 semi-quantitative RT-PCR을 이용하여 UVA 조사된 사람 진피 섬유아세포에 MMP-2 (gelatinase)의 활성 감소를 확인하였다. 결론적으로 권백은 자외선에 의한 세포손상을 보호하여 항노화 화장품의 새로운 소재로 이용될 것으로 사료된다.