• Title/Summary/Keyword: thymidine

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CHEMOSENSITIVITY OF CANCER CELLS TO ANTICANCER DRUGS USING DYE EXCLUSION ASSAY, [3H] THYMIDINE INCORPORATION, AND CLONOGENIC ASSAY (두경부악성종양세포주의 항암제감수성 시험에 관한 실험적 연구)

  • Jin, Woo-Jeong
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.15 no.1
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    • pp.35-48
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    • 1993
  • The in vitro predictive tests in cancer chemotherapy of cancer cell lines to anticancer drugs were determined using novel dye exclusion assay [NDEA], [3H] thymidine incorporation, and clonogenic assay [CA>. Antitumor effect of Bleomycin, Cis-platin, Vinblastine, Methotrexate to HEp-2, B16 cell lines using rapid assays was compared with [CA> in this study. In dye exclusion assay of B l6 cell line, cancer cells were sensitive to Bleomycin at all concentrations, to Vinblastine at the level of peak plasma concentration [PPC], ${\times}1/10$ [PPC](P<0.05). And Bleomycin revealed relatively good cytotoxicity than that of CDDP and vinblastine at ${\times}10$[PPC], (P<0.05). HEp-2 cells were resistive to methotrexate at the level of ${\times}100$[PPC] (P<0.05) In [3H] thymidine incorporation assay, B 16 cells were sensitive to Bleomycin, CDDP, Vinblastine at the level of [PPC], ${\times}10$ [PPC](P<0.01). Dose-dependent drugs of bleomycin, CDDP were more sensitive than Vinblastine at high concentration (P<0.05). In clonogenic assay, HEp-2 cell line was sensitive to three drugs of all concentrations except ${\times}10$ [PPC] of CDDP. B 16 cell line was sensitive to all drugs(P<0,01). In comparison of chemosensitivity tests among three assays, the results were correlated(${\gamma}=0.99$, P<0.05).

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The Glycopeptide, a Promoter of Thymidine Uptake, from Aloe Vera

  • Yang, Mi-Rim;Kang, Chun-Geun;Roh, Yeon-Suk;Son, Byeng-Wha;Choi, Hong-Dae;Park, Young-In;Lee, Seung-Ki;Choi, Sung-Won;Chung, Myung-Hee
    • Natural Product Sciences
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    • v.4 no.2
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    • pp.62-67
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    • 1998
  • As a part of search for new biologically active constituents from aloe, we have isolated a glycopeptide, called G1G1M1DI2, from the gel(G1) of Aloe vera. Chemical and spectroscopic evidence indicated that G1G1M1DI2 is a glycopeptide. The molecular weight of G1G1M1DI2 was about 5,500 daltons, and the carbohydrate and protein contents were 20.9% and 32.6%, respectively. Periodate oxidation and enzymic degradation gave peptide moiety and carbohydrate moiety, respectively. Carbohydrate moiety is composed of fucose, galactose, glucose and mannose in a molar ratio of 0.5:2.4;48.8:48.3. Peptide moiety is composed of fifteen amino acids, and glutamic acid and glycine were the major componants. The glycopeptide, G1G1M1DI2, stimulated thymidine uptake of SCC 13 cells about 6.5 times the control. This result suggests that this glycopeptide has a skin cell proliferating activity.

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Effects on Thymidine Analogs on Mitomycin C Induced DNA Repair Synthesis (Mitomycin C에 의한 DNA 回復合成에 미치는 Thymidine 相似體의 影響)

  • Park, Kyung-Hee;Park, Sang-Dai
    • The Korean Journal of Zoology
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    • v.20 no.2
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    • pp.93-99
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    • 1977
  • Dose response forDNA repair synthesis induced by various concentrations of MMC (0.05 $\\sim$ 0.5 $\\mu$g/ml) in HeLa $S_3$ cells was not dose-dependent and the amounts of it were relatively lower, representing $7\\sim9%$ of total DNA synthesizing cells in $0.1\\sim0.5 \\mug/ml$ concentrations. Time dependence study showed that MMC-induced DNA repair synthesis occurred as long as for 24 hours with similar incidences in all time courses. Pretreatment with BUdR was found to have a sensitization effect on MMC-induced DNA repair synthesis, but that with IUdR was not. Combined treatment with BUdR of IUdR and MMC suppressed remarkably the semiconservative DNA synthesis especially at later time course. These results seem to suggest that damages induced in DNA by MMC might be repaired by both fast and slow excision processes.

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Purification and Cloning of o Protein Secreted from Lactobacillus acidophilus

  • Han, Seo-Yeong;Lee, Yeong-Seon;Im, Jeong-Bin;Hwang, Deok-Su
    • Animal cells and systems
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    • v.2 no.3
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    • pp.355-359
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    • 1998
  • Among the proteins secreted from Lactobacillus acidophilus KCTC 3151, a 36 kDA and 24 kDa protein, whose amounts were relatively abundant, were purified and their N-terminal amino acid sequences determined. The N-terminal amino acid sequence of 36 kDa protein exhibited high homology with thymidine phosphorylase and glyceraldehyde-3-phosphate dehydrogenase. The N-terminal amino acid sequence of the 24 kDa protein did not show significant homology with proteins in Protein Data Base nor Gene Bank. Nucleotide sequence of the gene encoding 36 kDa protein indicates that the protein possesses the domains for a-helical, phosphate binding and pyrimidine binding sites, which are also shown in thymidine phosphorylases. Also, the protein contains conserved domains of dehydrogenase II and III. However, the activity of thymidine phosphorylase or glyceraldehyde-3-puospnate dehydrogenase could not be detected in the purified fractions of the 36 kDa protein.

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Photodynamic Therapy-Mediated Temporal Expression of Thymidine Kinase Genes Ligated to the Human Heat Shock Promotor: Preliminary in vitro Model Study of Enhanced Phototoxicity by PDT-Induced Gene Therapy

  • Kim, Mo-Sun;Kim, Tae-An;Kim, Jong-Ki
    • Journal of Photoscience
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    • v.9 no.3
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    • pp.41-43
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    • 2002
  • PDT-mediated cyototoxicity basically depends on the penetrated light-dose into the tumor tissue. This limits the efficiency of PDT to the superficial tumor region typically less than 1 cm. The localized photochemical generation of reactive oxygen species, including singlet oxygen is known to increase expression of assortment of early response genes including heat shock protein. In order to increase PDT cytotoxicity in the treatment of solid tumor, it is desirable to combine PDT with other therapeutic effects. In this preliminary study we evaluated enhanced cytotoxicity from the PDT-mediated expression of thymidine kinase in a transfected tumor cell line. Two types of photo sensitizers, a hematoporphyrin derivative(Photogem, Russia) and aluminium sulphonated phthalocyanine(Photosense, Russia) were used to evaluate the overexpression of hsp-70 in PDT-treated cell. Transient increase of hsp-70 was observed at 6-8 hrs later following irradiation in the photosense-treated cell whereas it was not observed in Photogem-treated cell. In the presence of ganciclovia, transfected cell showed a 17% increase in the cytotoxicity compared to the PDT only cell.

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Influence of Panax Ginseng on DNA Synthesis of Submandibular Gland in Mice Receiving ACTH(II) (고려인삼이 ACTH를 받은 마우스의 악하선 조직 DNA 합성능에 미치는 영향( II ))

  • Hong, Yong-Ha;Kim, Kee-Yun
    • The Korean Journal of Physiology
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    • v.8 no.2
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    • pp.45-48
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    • 1974
  • It was planned to investigate, in mice that received ACTH, the influence of Panax Ginseng upon DNA synthesis of submandibular gland by observing incorporation of $[^3H]$ thymidine into the tissue cells. Thirty male mice $(body\;weight:\;18{\sim}20\;g)$ were divided equally into the ginseng-ACTIH and the saline-ACTH groups. Each animal of the ginseng-ACTH and the saline-ACTH groups received every day (subcutaneously) 0.05 m1/10 g body weight of ginseng extract(4 mg of ginseng alcohol extract in 1 ml of saline) and the same amount of saline, respectively, for 5 days. On the 5th experimental day, all animals received 0.01 unit of ACTH intraperitoneally one hour after the last medication, and $1\;{\mu}Ci/g$ body weight of $[^3H]$ thymidine after one more hour. Five animals, at a time, of each group were sacrificed 1, 10, and 24 hr after $[^3H]$ thymidine administration, and the radioactivity of cells in their mandibular gland was measured autoradiographically in terms of the % number of radioactive cells in 1,000 cell counts (Radioactive Index, R.1.). Results obtained were as follows: 1. The radioactive indices obtained from submandibular gland of the saline-ACTH group 1, 10 and 24 hr after $[^3H]$ thymidine administration were $15.2{\pm}0.32,\;20.1{\pm}0.30,\;and\;24.5{\pm}0.52(mean{\pm}S.D.)$ in the mucous cells, $13.0{\pm}0.22,\;10.2{\pm}0.05,\;and\;7.5{\pm}0.42$ in the serous cells. and $10.5{\pm}0.40,\;13.6{\pm}0.32,\;and\;15.9{\pm}0.42$ in the duct cells, while the $mean{\pm}S.D.$ of the values obtained from the 3 cell types 1, 10 and 24 hr after $[^3H]$ thymidine were $10.9{\pm}0.28,\;12.4{\pm}0.31,\;and\;10.0{\pm}0.39.$ Thus the radioactive indices obtained from the ginseng-ACTH group were generally lower than those obtained from the saline-ACTH group. It is inferred from the above results that the ginseng tends to promote the suppressive action of ACTH upon DNA synthesis of cells in the mandibular gland.

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Cloning of Thymidine Kinase Gene of Herpes simplex Virus Type-1 (Herpes simplex Virus Type-1 Thymidine Kinase 유전자의 크로닝)

  • Kang, Hyun;Park, Kap-Joo;Cha, Sung-Chul;Kim, Soo-Yung;Yang, Ki-Sang;Kim, Nam-Joo;Lee, Hyung-Hoan
    • The Journal of Korean Society of Virology
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    • v.26 no.1
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    • pp.121-129
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    • 1996
  • Multiplication of Herpes simplex virus type-1 was observed by electronmicroscopy, a gene library of the genome was constructed and thymidine kinase gene was cloned. Vero cells infected with the virus were lysed 48 h p.j. and multinucleated giant cells were observed approximately at 72 h p.i. The nucleocapsids were observed in nuclei and cytoplasm, and the assembled nucleocapsids were budded out through the vacuole and cytoplasmic membranes, and then virions were released from the cells. HSV-1 genome DNA was digested with BamHI and BglII enzymes and then the gene library of the genome fragments were constructed. The BamHI cleaved the genome DNA into twenty-seven fragments in the range of 1.1 - 14 kb, and BglII cleaved the genome DNA into sixteen fragments in the range of $4.5{\sim}20.1\;kb$. The pHLA-12 and pHLB-4 recombinant plasmids were contained TK gene by Southern blot analysis. The molecular sizes of the fragments which contained the TK gene were 3.74 in pHLA-12 and 6.41kb in pHLB-4 recombinant plasmid, respectively.

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Interaction of $17{\beta}-Estradiol$ with EGF and IGF-I on Proliferation and $P_i$ Uptake in Primary Cultured Rabbit Renal Proximal Tubular Cells

  • Han, Ho-Jae;Lee, Yeun-Hee
    • The Korean Journal of Physiology and Pharmacology
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    • v.2 no.4
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    • pp.493-501
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    • 1998
  • The most significant direct role of estrogen in vivo is its ability to elicit receptor-mediated cellular proliferation in mammalian target tissues. However, the mechanism by which exogenously added estrogen causes the neoplastic transformation of renal cortical cells is yet to be uncovered. The present study was designed to evaluate interaction of $17{\beta}-estradiol\;(E_2)$ with epidermal growth factor (EGF) and insulin-like growth factor-I (IGF-I) on proliferation and $P_i$ uptake in primary cultured rabbit renal proximal tubular cells in phenol red-free, hormonally defined-medium. $[^3H]-thymidine$ incorporation increased markedly by about 133% and 141% more in the presence of $10^{-9}\;and\;10^{-6}\;M\;E_2$, respectively, than that of control. Cell count was 162% and 143% greater in the presence of $10^{-9}\;and\;10^{-6}\;M\;E_2$ , respectively, compared with control. Among all time points examined, there was an increase in $[^3H]-thymidine$ incorporation in the presence of $10^{-9}\;M\;E_2$ at day 9 or 13, respectively. However, $E_2$ ($10^{-9}\;M$) significantly drove up cell count to 160% of that of control at day 13, while it had a slight but statistically insignificant effect at day 9. $E_2-induced$ stimulation of $[^3H]-thymidine$ incorporation was completely reversed by $E_2$ antagonists (progesterone or tamoxifen). $E_2$ ($10^{-9}\;M$) or EGF ($10^{-8}\;M$) significantly stimulated $[^3H]-thymidine$ incorporation by 144% and 154% of control. $E_2$ plus EGF was synergistic on $[^3H]-thymidine$ incorporation (204% of control), while $E_2$ plus IGF-I showed a slight but no significant synergistic effect. Cell number also displayed similar pattern. $E_2$ ($10^{-9}\;M$) significantly stimulated $P_i$ uptake to 134% of control. $E_2$-induced stimulation of $P_i$ uptake was partially reversed by $E_2$ antagonists. EGF or IGF-I ($10^{-8}\;M$) significantly also increased $P_i$ uptake to 132% or 129% of control. $E_2$ plus EGF had synergistic effect on $P_i$ uptake, while $E_2$ plus IGF-I did not. In conclusion, $E_2$ may act not only directly interaction with its receptors but also indirectly as a modulator of EGF in proliferation and $P_i$ uptake of primary cultured rabbit renal proximal tubular cells.

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Effects of Insulin-like Growth Factor in Peritoneal Fluid of Patients with Endometriosis on the Proliferation of Endometrial Stromal Cells (자궁내막증 환자의 복강액내 IGF가 자궁내막 기질세포 증식에 미치는 영향)

  • Kim, Jung-Gu;Suh, Chang-Seok;Kim, Seok-Hyun;Choi, Young-Min;Moon, Shin-Yong;Lee, Jin-Yong
    • Clinical and Experimental Reproductive Medicine
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    • v.26 no.3
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    • pp.331-338
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    • 1999
  • The purposes of this study were to evaluate the effects of insulin-like growth factor (IGF)s in peritoneal fluid (PF) from patients with and without endometriosis on the proliferation of endometrial stromal cells and to investigate the effects of type I IGF receptor antibody on the response of endometrial stromal cells to PF from patients with endometriosis. IGFs in PF from patients with endometriosis (n=14) and without endometriosis (n=10) were measured by immunoradiometric assay and PF samples were divided into low IGF-I PF group (less than 85 ng/ml) and high IGF-I PF group (more than 85 ng/ml). Endometrial stromal cells from patients without endometriosis were cultured in serum free media in the presence or absence of 1 % PF and thymidine incorporation test were used to evaluate the proliferation of endometrial stromal cells. Also cultures were incubated with type I IGF receptor monoclonal antibody (${\alpha}IR_3$) before adding PF. PF from patients with endometriosis and without endometriosis increased thymidine incorporation in endometrial stromal cells. In patients with endometriosis, high IGF-I PF group had high IGF-II levels and resulted in higher thymidine incorporation than low IGF-I PF group but no significant difference in increase in thymidine incorporation between high IGF-I and low IGF-I PF group was noted in patients without endometriosis. There was not a significant correlation between increase in thymidine incorporation and IGF-I levels in PF from patients without endometriosis but in PF from patients with endometriosis. Preincubation with ${\alpha}IR_3$ significantly inhibited the mitogenic response of endometrial stromal cells to PF. Our data indicate that IGF-I in PF may be involved in the growth of ectopic endometrium in patients with endometriosis.

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