• Title/Summary/Keyword: DNA Synthesis

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Caffeic Acid Phenethyl Ester Inhibits Cell Proliferation and Induces Apoptosis in Human Ovarian Cancer Cells

  • Park, Hyung-Joo;Yang, Seung-Joo;Mo, Jin-Young;Ryu, Geun-Chang;Lee, Kyung-Jin
    • Korean Journal of Environmental Biology
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    • v.28 no.4
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    • pp.196-201
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    • 2010
  • The phenethyl ester of caffeic acid (CAPE), an active component of honeybee propolis extract, is shown to inhibit cancer growth previously. However, studies on human ovarian cancer are largely obscure. This study evaluated the effects of CAPE as a potential anti-proliferative and pro-apoptotic agent in the human ovarian cancer line, OVCAR-3. CAPE treated OVCAR-3 cells showed inhibition of cell viability and proliferation in a dose-dependent manner by WST-1 assay, LDH assay and bromodeoxyuridine (BrdU) incorporation assay. Furthermore, CAPE-mediated OVCAR-3 cell growth inhibition was associated with apoptotic changes as evident by cell cycle arrest and accumulation of cells in the apoptotic phase and DNA fragmentation. Taken together, CAPE inhibits cell proliferation via DNA synthesis reduction and induces apoptotic cell death via DNA damage, thus elucidating a novel, plausible mechanism of CAPE anti-tumorigenic property in OVCAR-3 cells.

cDNA Sequence and mRNA Expression of a Putative Glutathione S-Transferase from the Mole Cricket, Gryllotalpa orientalis

  • Kim, Iksoo;Lee, Kwang-Sik;Jin, Byung-Rae;Kim, Jin-Won;Ryu, Kang-Sun;Ahn, Mi-Young
    • International Journal of Industrial Entomology and Biomaterials
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    • v.6 no.2
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    • pp.157-162
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    • 2003
  • The glutathione S-transferase (GSTs) are enzymes responsible for the protection of cells from chemical toxicants and oxidative stress. We describe here the cDNA sequence and mRNA expression of a putative GST from the mole cricket, Gryllotalpa orientalis. The G. orientalis GST cDNA sequences comprised of 621 bp encoding 207 amino acid residues. The multiple sequence alignment of G. orientalis GST gene with other known insect GSTs showed several conserved residues that may be essential for the enzymatic activity of the protein. Phylogenetic analysis of the deduced amino acid sequences of G. orientalis GST gene with other insect GST sequences revealed that the G. orientalis GST gene belongs to class I GST, forming a strong monophyletic group (100% bootstrap value) exclusively for class I GSTs from a diverse insect species. Northern blot analysis confirmed midgut-specific expression at transcriptional level, evidencing the midgut as a site for GST synthesis.

Effect of Gleditsiae Spina on Proliferation of Transplanted-L1210 cells in Mice (조각자가 생쥐에 이식된 L1210 세포의 증식에 미치는 영향)

  • 조선경;은재순;김대근;소준노;오찬호;송정모
    • The Journal of Korean Medicine
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    • v.22 no.4
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    • pp.37-44
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    • 2001
  • Objectives : Cellular death by apoptosis is an active process, depending on gene transcription and protein synthesis. It was reported that nitric oxide can induce apoptosis in several cancer cell-lines. We have previously observed that proliferation of Ll210 cells was inhibited by the administration of Gleditsiae Spina water extract (GE). In this present study, the mechanism of inhibitory action on the proliferation of L l210 cells was examined. Methods : The cell proliferation was determined by MTT assay and DNA fragmentation was determined by a flow cytometry. Results : The administration of GE decreased proliferation of L1210 cells and enhanced DNA fragmentation in vivo system. DNA fragmentation of L1210 cells was enhanced by co-culture of peritoneal macrophages obtained from GE-administered mice in vitro and it was partly inhibited by L-NMMA, nitric oxide synthetase inhibitor. In addition, GE increased nitric oxide production from peritoneal macrophages of L1210-transplanted mice. Conclusions : These results suggest that the inhibitory action of GE on proliferation of transplanted-L1210 cells is partly caused by an induction of apoptosis via production of nitric oxide in macrophages.

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Cloning and Sequence Analysis of Ribosomal Protein S4 cDNA from Root of Panax ginseng

  • In Jun-Gyo;Lee Bum-Soo;Song Won-Seob;Bae Chang-Hyu;Choi Seong-Kyu;Yang Deok-Chun
    • Plant Resources
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    • v.8 no.2
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    • pp.110-115
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    • 2005
  • Ribosomal protein complex with ribosomal RNA to form the subunits of the ribosome serve essential functions in protein synthesis. A full-length cDNA (PRPS4) encoding ribosomal protein S4 has been isolated and its nucleotide sequence determined in ginseng plant (Panax ginseng). A PRPS4 cDNA is 1105 nucleotides long and has an open reading frame of 792 bp with a deduced amino acid sequence of 264 residues (pI 10.67). The deduced amino acid sequence of PRPS4 matched to the previously reported ribosomal protein S4 genes. Their degree of amino acid identity ranged from 68 to $92\%$. Phylogenetic analysis based on the amino acid residues showed that the PRPS4 grouped with ribosomal protein S4 of S. tuberosum (CAA54095).

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Prospect of plant molecular cytogenetics in the 21st century

  • Mukai, Yasuhiko
    • Proceedings of the Korean Society of Life Science Conference
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    • 2003.10a
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    • pp.14-27
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    • 2003
  • The genomes of Arabidopsis and rice have been fully sequenced. Genomic sequencing provides global information about genome structure and organization. A comprehensive research account of our recent studies conducted on genome painting, comparative genomics and genome fusion is provided in order to project the prospects of plant cytogenetic research in post-genomics era. Genome analysis by GISH using genome painting is demonstrated as an excellent means suitable for visualization of a whole genome, since total genomic DNA representing the overall molecular composition of the genome is used as a probe. FISH on extended DNA fibers has been developed for high-resolution FISH and has contributed to determining the copy number and order of genes. We have also mapped a number of genes involving starch synthesis on wheat chromosomes by FISH and compared the position of these genes on linkage map of rice. Macro synteny between wheat and rice can be observed by comparing the location of these genes in spite of the fact that the size of DNA per chromosome differs by 20 fold in two. Moreover, to approach our goal towards making bread and udon noodles from rice flour in future by incorporating bread making and the noodle qualifies in rice, we have been successful in introducing large genomic DNA fragments containing agronomically important genes of wheat into a rice by successive introduction of large insert BAC clones, there by expanding genetic variability in rice. We call this method genome fusion.

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A Novel UV-Sensitivity Mutation Induces Nucleotide Excision Repair Phenotype and Shows Epistatic Relationships with UvsF and UvsB Groups in Aspergillus nidulans

  • Baptista, F.;Castro-Prado, M.A.A.
    • Journal of Microbiology
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    • v.39 no.2
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    • pp.102-108
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    • 2001
  • DNA damage response has a central role in the maintenance of genomic integrity while mutations in related genes may result in a range of disorders including neoplasic formations. The uvsZl characterized in this report is a navel uvs mutation in Aspergillus nidulans, resulting in a nucleotide excision repair (NER) phenotype: UV-sensitivity before DNA synthesis (quiescent cells), high UV-induced mutation frequency and probable absence of involvement with mitotic and meiotic recombinations. The mutation is recessive and nan-allelic to the previously characterized uvsA101 mutation, also located on the paba-y interval on chromosome I. uvsZl skewed wild-type sensitivity to MMS, which suggests non-involvement of this mutation with BER. Epitasis tests showed that the uvsZ gene product is probably involved in the same repair pathways as UVSB or UVSH proteins. Although mutations in these proteins result in an NER phenotype, UVSB is related with cell cycle control and UVSH is associated with the post-replicational repair pathway. The epistatic interaction among uvsZl and uvsB413 and uvsH77 mutations indicates that different repair systems may be related with the common steps of DNA damage response in Aspergillus nidulans.

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Functional Implications in Apoptosis by Interferon Inducible Gene Product 1-8D, the Binding Protein to Adenovirus Preterminal Protein

  • Joung, In-Sil;Angeletti, Peter C.;Engler, Jeffrey A.
    • Journal of Microbiology
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    • v.41 no.4
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    • pp.295-299
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    • 2003
  • Adenovirus (Ad) precursor to the terminal protein (pTP) plays an essential roles in the viral DNA replication. Ad pTP serves as a primer for the synthesis of a new DNA strand during the initiation step of replication. In addition, Ad pTP forms organized spherical replication foci on the nuclear matrix (NM) and anchors the viral genome to the NM. Here we identified the interferon inducible gene product 1-8D (Inid) as a pTP binding protein by using a two-hybrid screen of a HeLa cDNA library. Of the clones obtained in this assay, nine were identical to the Inid, a 13-kDa polypeptide that shares homology with genes 1-8U and Leu-13/9-27, most of which have little known functions. The entire open reading frame (ORF) of Inid was cloned into the tetracycline inducible expression vector in order to determine the biological functions related with adenoviral infection. When Inid was introduced to the cells along with adenoviruses, fifty to sixty percent of Ad-infected cells expressing Inid had rounded morphology, which was suggestive of apoptosis. Results from the terminal deoxynucleotidyl transferase (TdT) and DNA fragmentation assays confirmed that Inid induces apoptosis in Ad-infected or in uninfected cells. The Inid binding to pTP may target the cell for apoptotic destruction as a host defense mechanism against the viral infection.

THE STUDY ON EFFECTS OF THE PLATELET-DERIVED GROWTH FACTOR-AA, BB ON THE CELLULAR ACTIVITY OF THE HUMAN PERIODONTAL LIGAMENT CELLS (Platelet-derived growth factor-AA, BB가 치주인대세포의 세포활성에 미치는 영향에 대한 연구)

  • Oh, Sang-Deok;Lee, Jae-Mok;Suh, Jo-Young
    • Journal of Periodontal and Implant Science
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    • v.24 no.2
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    • pp.303-320
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    • 1994
  • Current acceptable methods For promotin gperiodontal regeneration are base on removal of diseased soft tissue, root treatment, guided tissue regeneration, inteoduction of new graft materials and biological mediators. Platelet-derived growth factor(PDGF) is one of polypeptide growth factor. PDGF has been reported as a biological mediator which regulates activities of wound healing process including the cell proliferation, migration and metabolism. The purposes of this study is to evaluate the effects of PDGF-AA, BB on the periodontal ligament cells to use as a regeneration promoting agent of periodontal tissue. Human periodontal ligament cells were prepared from the first premolar tooth extracted for the orthodontic treatment and were cultured in DMEM/10% FBS at the $37^{\circ}C$, 5% $CO_2$ incubator. Author measured the DNA synthesis, total protein, collagen and noncollagenous protein synthesis and alkaline phosphatase activity according to the concentration of PDGF-AA and BB(0, 0.1, 1, 10, 100ng/ml). The results were as follows : The DNA synthetic activity was increased dose dependently by PDGF-AA and BB. The maximum mitogenic effect was at the 100ng/ml of PDGF-AA and 10ng/ml of PDGF-BB. The total protein, collagen and noncollagen systhesis was increased dose dependently by PDGF-AA and BB. The % of collagen was slightly decresed according to the concentration of PDGF-AA and BB. The effect of PDGF-AA and BB were not specific for collagen synthesis since it also increased noncollagenous protein synthesis. The effect of PDGF-AA and BB on alkaline phosphatase activity did not show any significant, meanwhile the alkaline phosphatase activity of 14 days group showed significnat increase. In conclusion, PDGF-AA and BB may have important roles in stimulation of DNA synthesis in human periodontal ligament cells, which means an increase in collagen-synthesizing cells, and may be useful for clinical application in periodontal regenerative procedures.

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cDNA Microarray Analysis of Differential Gene Expression in Boar Testes during the Prepubertal Period

  • Lee, Dong-Mok;Lee, Ki-Ho;Choi, Jin Ho;Hyun, Jin Hee;Lee, Eun Ju;Bajracharya, Prati;Lee, Yong Seok;Chang, Jongsoo;Chung, Chung Soo;Choi, Inho
    • Asian-Australasian Journal of Animal Sciences
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    • v.22 no.8
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    • pp.1091-1101
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    • 2009
  • In an attempt to understand the biochemical mechanism for the synthesis of the anabolic steroid, 19-nortestosterone, produced by prepubertal boar testes and its physiological role, normalized complementary DNA (cDNA) from boar testes was generated. DNA sequencing of 2,016 randomly selected clones yielded 794,116 base pairs of high quality nucleotide sequence. Computer-assisted assembly of the nucleotide sequence of each clone resulted in 423 contigs and 403 singletons including several genes for steroidogenic enzymes and molecules related to steroid metabolism. Analysis of gene expression pattern by use of the presently-fabricated cDNA microarray identified a number of genes that were differentially expressed during the postnatal development period in boar testes. Two genes of unknown function were identified to be highly expressed in the testis of 2-weeks-old neonatal boar. In addition, the sequencing of open reading frames of these genes revealed their homology with human alpha hemoglobin and Homo sapiens hypothetical LOC643669, transcript variant 1. Moreover, the transcripts of these genes were also detected in porcine muscle and adipocytes, in addition to Leydig cells of pigs.

Quantitative analysis of gene expression pattern in aspergillus nidulans mycelia by sequencing of 3-directed cDNA clones

  • Park. Yoon-Dong;Lee, Dong-Whan;Lee, Seog-Jae;Kim, Jong-Hwa;Chae, Keon-Sang
    • Journal of Microbiology
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    • v.34 no.1
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    • pp.25-29
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    • 1996
  • Since sequencing of randomly selected cDNA clones has been known to be a powerful approach to obtain information on gene expression pattern in specific cells or tissues, we have analyzed a 3'-directed cDNA library of vegetative mycelia of A. nidulans by single-pass sequencing of hundreds of randomly selected clones. Sequencing of 292 cDNA clones yielded 209 gene signatures (GSs) probably representing highly or lesser expressed genes in the vegetative mycelia. Among the 209 GSs, 25 (79 cDNA clones) appeared more than once and 184 only once. One GS appeared at a highest frequency of 6 times, 2 GSs5 times, 4 GSs 4 times, a GSs 3 times and 16 GSs twice. About 6.6% GSs comprizing of 13 GSs showed alternative polyadenylation. Among 23 redundant GSs, three were common in both mycelia and sexual organs, and 22 were probably mycelia-specific. Out of 209 GSs, 36 were identified in GenBank showing of 70% or greater similaritis. Only six GSs were for A. nidulans genes, and 13 GSs were of DNA or genes encoding cytoplasmic or organellar proteins. This pattern is similar to those in the human HepG2 cell line and in human colonic mucosa, although very few genes for nuclear proteins and for protein synthesis were in A. nidulans.

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