• 제목/요약/키워드: DNA-dependent

검색결과 1,347건 처리시간 0.025초

실험실 내에서 배양된 편평상피암 세포주에 대한 염화불소의 효과 (THE EFFECTS OF SODIUM FLUORIDE ON SQUAMOUS CELL CARCINOMA CELL LINE CULTURED IN VITRO)

  • 박노부
    • Maxillofacial Plastic and Reconstructive Surgery
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    • 제18권2호
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    • pp.316-322
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    • 1996
  • 불소의 유전물질에 대한 독성효과는 현재까지 논란의 대상이되고 있으며, 서로 상반된 연구 결과들이 보고되고 있다. 저자는 불소의 세포 유전학적 효과를 규명하고자 실험실 내에서 배양된 편평상피암 세포주에 염화불소를 처리하여 세포독성 검사와 염색체이상 검사를 시행하여 다음과 같은 결론을 얻었다. 1. 염화불소의 농도와 처리시간에 비례하여 세포성장이 현저히 감소하므로 염화불소는 편평상피암 세포주에 독성을 가진다. 2. 염화불소의 농도가 증가할수록 염색체이상 발현빈도가 현저히 높게 나타나므로 염화불소는 편평상피암 세포주에서 DNA 손상을 야기할 수 있다. 염색체 이상의 형태는 chromatid break가 가장 많이 나타났다.

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염기서열과 PCR-Restriction Fragment Length Polymorphism 분석에 의한 Mycobacteria 동정 (Identification of Mycobacteria by Comparative Sequence Apalysis and PCR-Restriction Fragment Length Polymorphism Analysis)

  • 국윤호
    • 대한미생물학회지
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    • 제34권6호
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    • pp.561-571
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    • 1999
  • Diagnosis of mycobacterial infection is dependent upon the isolation and identification of causative agents. The procedures involved are time consuming and technically demanding. To improve the laborious identification process mycobacterial systematics supported by gene analysis is feasible, being particularly useful for slowly growing or uncultivable mycobacteria. To complement genetic analysis for the differentiation and identification of mycobacterial species, an alternative marker gene, rpoB encoding the ${\beta}$ subunit of RNA polymerase, was investigated. rpoB DNAs (342 bp) were amplified from 52 reference strains of mycobacteria including Mycobacterium tuberculosis H37Rv (ATCC 27294) and clinical isolates by the PCR. The nucleotide sequences were directly determined (306 bp) and aligned using the multiple alignment algorithm in the MegAlign package (DNASTAR) and MEGA program. A phylogenetic tree was constructed with a neighborhood joining method. Comparative sequence analysis of rpoB DNA provided the basis for species differentiation. By being grouped into species-specific clusters with low sequence divergence among strains belonging to same species, all the clinical isolates could be easily identified. Furthermore RFLP analysis enabled rapid identification of clinical isolates.

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에틸카바메이트 대사산물에 의해 유발된 마우스 피부 종양에 대한 홍삼의 억제효과 (Inhibitory Effects of Red Ginseng on Skin Tumor Formation Induced by Ethyl Carbamate Metabolites)

  • 박광균;오상환;정원윤
    • Toxicological Research
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    • 제16권1호
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    • pp.9-16
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    • 2000
  • Ginseng (the root of Panax ginseng C. A. Meyer, Araliaceae) has been used for traditional medicine in China, Korea, Japan and other Asian countries. It is most often used as a general tonic, and it involves a wide range of pharmacological actions, such as antiaging, adaptogen-like effect to foreign deleterious infringement, immunoenhancement, antistress, antitumor, and antioxidant actions. Red ginseng showed anticarcinogenic activity against various chemical carcinogens in mouse and cancer-preventive effect of human being as on mice in experimental and epidemiological studies. In the present study, we have found the protective properties of red ginseng against vinyl carbamate (VC) which is the proximate carcinogen of ethyl carbamate and its ultimate carcinogenic epoxides. Red ginseng exhibited dose-dependent inhibition on the mutagenci activities of boty VC in the presence of S9 mix and vinyl carbamate epoxide (VCO) without metabolic activation in Salmonella typhimurium TA1535. Formation of DNA adducts from VCO was also attenuated in the presence of red ginseng. Oral administration of red ginseng prior to the topical application of each of the above carcinogens and TPA treatment resulted in significant reduction in both incidence and multiplicity of skin tumors in mice. These results indicate that red ginseng possesses a strong chemopreventive effect against mouse skin carcinogenesis induced by VC or VCO.

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Nucleotide Insertion Fidelity of Human Hepatitis B Viral Polymerase

  • Kim, Youn-Hee;Hong, Young-Bin;Suh, Se-Won;Jung, Gu-Hung
    • BMB Reports
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    • 제33권2호
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    • pp.126-132
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    • 2000
  • The hepadnaviruses replicate their nucleic acid through a reverse transcription step. The MBP-fused HBV polymerase was expressed in E. coli and purified by using amylase affinity column chromatography. The purified protein represented DNA-dependent DNA polymerase activity. In this report, the MBP-HBV polymerase was shown to lack 3'$\rightarrow$5' exonuclease activity, like other retroviral RTs. The ratio of the insertion efficiency for the wrong versus right vase pairs indicates the misinsertion frequency (f). The nucleotide insertion fidelity (1/f), observed with the MBP-HBV polymerase and HIV-1 RT, was between 60 and 54,000, and between 50 and 73,000, respectively, showing that they are in close range. A relatively efficient nucleotide incorporation by the MBP-HBV polymerase was observed with a specificity of three groups: (1) A : T, T : A>C : G, G : C (matched pairs), (2) A : C, C : A>G: T, T : G (purine-pyrimidine and pyrimidine-purine mispairs), and (3) C : C, A : A, G : G, T : T>T : C, C : T>A : G, G : A (purine-purine or pyrimidine-pyrimidine mispairs), and their order is (1)>(2)>(3). The data from the nucleotide insertion fidelity by the MBP-HBV polymerase suggest that the HBV polymerase may be as error-prone as HIV-1 RT.

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HepG2 간암세포에 대한 부자 추출물의 고사 유도 효과 (The Apoptosis-inducing Effect of Radix Aconiti Extract in HepG2 Human Hepatoma Cells)

  • 권강범;김은경;정은실;심정섭;김강산;신병철;송용선;류도곤
    • 대한한의학회지
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    • 제25권2호
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    • pp.33-40
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    • 2004
  • Objective : This study investigated the apoptotic effect and its mechanism of Radix Aconiti (RA) extract and aconitine, which is a major constituent of RA, in HepG2 human hepatoma cells. Methods : We used MTT and DNA fragmentation assay to investigate cell viability and apoptotic effect on RA extract-treated HepG2 cells. In addition, to clarify the mechanism of RA extract-induced apoptosis, we applied caspase-3 enzyme activity assay and Western blotting method on poly-(ADP-ribose) polymerase (PARP) protein expression. Results : Treatment with RA extract resulted in the decrease of cell viability, and this effect was caused from apoptosis as confirmed by discontinuous fragmentation of DNA in HepG2 cells, but aconitine did not. Also, RA extract-treated HepG2 cells induced the activation of caspase-3 enzyme activity in time- and dose-dependent manners, which was accompanied by the cleavage of 116 kD PARP to 85 kD product. Conclusions : These results suggest that the apoptotic effects of RA extract on HepG2 cells could not be explained by aconitine. Additionally, RA extract induced apoptosis in hepatoma cells through caspase-3 activation and subsequent PARP cleavage.

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가미삼출삼자탕의 항암활성에 관한 연구 (Study on Study on Antitumor Activity of Kamisamchulsamja-tang)

  • 김성언;이효정;김동희;송규용;김성훈
    • 동의생리병리학회지
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    • 제16권4호
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    • pp.667-673
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    • 2002
  • The purpose of this study was to investigate the effect of Kamisamchulsamja-tang (KSST) water extract on the antitumor activity. The results were summarized as follows: KSST extract exhibited a weak cytotoxicity against HT1080, A549, SK-OV-3, B16-F10 and SK-MEL-2 cells. KSST extract showed a inhibitoty effect on DNA topoisomerase I from calf thymus in a dose-dependent manner. Also, KSST extract showed antiadhesive effect on HT1080 cells but didn't showed on A549 cells to complex extracellular matrix. In pumonary colonization assay, a number of colonies in the lungs were decreased significantly in KSST treated group as compared with control group. In vitro neovascularization assays, angiogenesis was significantly inhibited in KSST treated group than control group. In CAM assay, KSST extract inhibited angiogenesis significantly at 15㎍/egg concentration as compared with control. From the above results it was concluded that KSST showed antitumor effect through the antimetastatic effect. So it is expected to be clinically helpful on the prevention of metastasis of cancer.

췌장암 세포주 MIA PaCa-2에서 황백 물 추출물에 의한 Apoptosis 유도 및 작용기전 (Apoptotic Effect of Phellodendri Cortex Water Extract on MIA PaCa-2 Cells)

  • 이인영;정황산;원진희
    • 동의생리병리학회지
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    • 제27권2호
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    • pp.202-211
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    • 2013
  • The purpose of this study is to investigate the apoptotic effect of Phellodendri Cortex water extract (PCWE) on pancreatic cancer cells and to find out the regulating mechanisms. Human-derived pancreatic cancer cell line, MIA PaCa-2 cells were treated by PCWE with various concentrations and the cytotoxicity was determined by MTT assay. The activation of Annexin V, DNA fragmentation, cell cycle arrest and caspase activation were observed to investigate the role of PCWE in pancreatic cancer cells. Also, to find out the regulating mechanisms, we examined the ROS production. The treatment of PCWE induced the cell death in both concentration and time dependent manner. The treatment of PCWE also increased the expression of Annexin V, DNA fragmentation, cell cycle arrest, and cleavage of caspase, which means cell-death PCWE induced was apoptosis but not necrosis. The ROS production was increased by PCWE treatment and the blockade of ROS inhibited the PCWE-induced cell death. These results could suggest that PCWE induced apoptosis via ROS release in pancreatic cancer cell.

Identification of the Genes Involved in the Fruiting Body Production and Cordycepin Formation of Cordyceps militaris Fungus

  • Zheng, Zhuang-Li;Qiu, Xue-Hong;Han, Ri-Chou
    • Mycobiology
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    • 제43권1호
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    • pp.37-42
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    • 2015
  • A mutant library of Cordyceps militaris was constructed by improved Agrobacterium tumefaciens-mediated transformation and screened for degradation features. Six mutants with altered characters in in vitro and in vivo fruiting body production, and cordycepin formation were found to contain a single copy T-DNA. T-DNA flanking sequences of these mutants were identified by thermal asymmetric interlaced-PCR approach. ATP-dependent helicase, cytochrome oxidase subunit I and ubiquitin-like activating enzyme were involved in in vitro fruiting body production, serine/threonine phosphatase involved in in vivo fruiting body production, while glucose-methanol-choline oxidoreductase and telomerase reverse transcriptase involved in cordycepin formation. These genes were analyzed by bioinformatics methods, and their molecular function and biology process were speculated by Gene Ontology (GO) analysis. The results provided useful information for the control of culture degeneration in commercial production of C. militaris.

Lactobacillus casei 의 Phospho-$\beta$-galactosidase 유전자의 지도작성과 Escherichia coli 내에서의 발현 (Mapping of Gene Encoding Phospho-$\beta$-galactosidase from Lactobacillus casei and its Expression in Escherichea coli)

  • 박정희;문경희;민경희
    • 미생물학회지
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    • 제30권6호
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    • pp.539-545
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    • 1992
  • Lactobacillus casei SM-M1 의 플라스미드로부터 phospho-$\beta$-galactosidase gene 을 갖는 DNA 를 E. coli 에 클로닝한 pPLac15(13kb) 의 재조합 플라스미드를 제조하였다.(15). pPLac15 DNA 를 분리하여 제한효소로 처리하여 제한효소 지도를 작성하였다. Phospho-$\beta$-galactosidase 유전자의 발현을 높이기 위하여 lac promoter 를 가진 pUC18 의 PstI 위치에 클닝하여 pPLac18 을 제조하였으며, 이것을 다시 EcoRI 으로 절단하여 pUC 18 에 클로닝하여 얻은 pPLac23 (7.6 kb) 를 얻었다. Phospho-$\beta$-galactosidase 효소활성은 pPLac23 의 형질전환주인 E. coli SW-23 에서는 pPLac15 를 가진 형질전환주인 E. coli SW-15 보다 약 1.8 배의 효소의 활성을 나타내었으며 pPLac18 을 가진 E. coli SW-18 보다는 약간 높은 활성을 나타내었다.

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배추김치 분획물의 in vitro 항암효과 (Invitro Anticancer Effect of Chinese Cabbage Kimchi Fractions)

  • 박건영;조은주;이숙희;강갑석
    • 한국식품영양과학회지
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    • 제28권6호
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    • pp.1326-1331
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    • 1999
  • In vitro anticancer effect of Chinese cabbage kimchi fractions was investigated by using human cancer cells, AGS human gastric adenocarcinoma cells and HT 29 human colon adenocarcinoma cells. The Chinese cabbage kimchi(fermented for 4 days at 15oC) was fractionated into 7 groups, methanol extract, hexane fraction(fr.), methanol soluble fr., dichloromethane fr., ethylacetate fr., butanol fr. and aqueous fr.. Chinese cabbage kimchi fractions inhibited the growth of AGS and HT 29 cancer cells as dose dependent. In particular, the dichloromethane fr. showed the highest inhibitory effect among other fractions. When the dichloromethane fr.(0.2mg/ml) was treated, the number of AGS and HT 29 survival cancer cells reduced to 12$\times$104/ml and 11$\times$104/ml compared to 166$\times$104/ml and 50$\times$104/ml of the controls, respectively. Chinese cabbage kimchi fractions also inhibited the DNA synthesis of the cancer cells. They inhibited the DNA synthesis of AGS human gastric adenocarcinoma cells more efficiently than that of HT 29 human colon adenocarcinoma cells. These results indicate that Chinese cabbage kimchi fractions show in vitro anticancer activity and the dichloromethane fr. among them reveals the highest effect.

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