• 제목/요약/키워드: thymidine incorporation

검색결과 158건 처리시간 0.025초

Ag-NORs 염색법과 [3H]thymidine incorporation법에 의한 caffeine의 NIH3T3 세포증식 억제효과 비교측정 (Comparison of Ag-NORs stain and [3H]thymidine incorporation in antiproliferative effect of caffeine on NIH3T3 cells)

  • 김성호;이차수
    • 대한수의학회지
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    • 제30권4호
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    • pp.459-464
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    • 1990
  • Inhibitory effect of caffeine on NIH3T3 cell proliferation was studied by using [$^3H$]thymidine incorporation and a modified one-step silver staining technique. The latter technique shows argyrophilic nucleolar organizer region-associated proteins (Ag-NORs), which are seen in nuclei as black dots. The result was compared with the counts of [$^3H$] thymidine incorporation. The results were summarized as follows; 1. The Ag-NORs numbers of NIH3T3 cells were $6.81{\pm}1.38$ at 24 hrs, $7.13{\pm}1.26$ at 48 hrs, $8.50{\pm}2.04$ at 72 hrs after incubation. 2. The numbers of Ag-NORs were significantly decrease in caffeine treated groups (p<0.01). 3. The counts of [$^3H$] thymidine incorporation were similar to the result of using Ag-NORs staining technique. It is concluded that Ag-NOR is a rapid, simple and compatible method to quantitate cell proliferation as compared with [$^3H$]thymidine incorporation.

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호수 생태계에서 세균 생산량 측정 방법의 비교 (Comparison of Methods for the Measurement of Bacterial Production in a Lake Ecosystem)

  • 김명운;강찬수;김상종
    • 미생물학회지
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    • 제28권4호
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    • pp.318-323
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    • 1990
  • The bacterial secondary production was measured at 6 sites of Lake Soyang in October, 1989 by $^{3}$H-thymidine incorporation rate. Verfication for the method of bacterial secondary production measurement showed that $^{3}$H-thymidine incorporated into DNA, RNA and protein by average percentage of 38.45, 42.27 and 20.07%, respectively. THe more increased incoporated $^{3}$H-thymidine, the more increasde DNA fraction, but protein fraction was generally low. Incorporation of rate of /usp 3/H-thymidine. $^{3}$H-leucine into protein correlated with protein fraction of incorporated $^{3}$H-thymidine. Conversion factors were calculated as follows; $1.83*10 ^{20}$ cells/moles of thymidine incorporated/hr and 1.69*10$^{22}$ cells/moles of leucine incorporated/hr.

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흰쥐의 소장 점막 세포의 성장에 미치는 Glutamine, Glycine과 Nucleosides/Nucleotide 혼합물의 효과 (Effects of Glutamine, Glycine and Nucleosides/Nucleotide Mixture on Intestinal Mucosal Growth in Rats)

  • 이선영;오현인
    • 한국식품영양과학회지
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    • 제26권1호
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    • pp.130-136
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    • 1997
  • 250g내외의 횐쥐에게 Thiry-Vella fistula를 만든 후 수술과 비경구 영양 효과로 인하여 위축되는 소장의 누공에 각각 nucleosides/nucleotide혼합용액 (Nuc군)이나 glutamine(Gln군), glycine(Gly군)등을 투여하여 소장 점막의 위축 방지나 재생효과가 있는지를 알아보고 자 실험하여 다음과 같은 결과를 얻었다. Thiry-Vella fistula 시술 후 9일 동안 2일에 한 번씩 총 4회 소장 누공으로 주입한 이미노산들과 nucleosides/nucleotide 혼합물의 효과는 누공의 소장 점막에 미치는 직접적인 효과와 비누공 부분에 대한 간접적인 효과로 나누어 볼 수 있다. Nucleosides/nucleotide 혼합물은 소장 누공으로 주입되어 직접 접촉되는 부분의 소장 DNA함량을 유의적으로 증가 시켰으며 단백질 함량도 높은 수치를 보여 주었고 조직 검사 결과 소장 융모의 길이와 표면적을 증가시킨 것으로 나타났다. 그러나 [$^3$H] thymidine incorporation의 결과는 대조군과 유의적인 차이를 보이지 않았다. Glycine은 소장 단백질과 DNA에 있어서 대조군보다는 높고 Nuc군보다는 낮은 경향을 보여 주었으나 [$^3$H]-thymidine incorporation은 누공과 비누공부분에서 모두 가장 높게 나타났으며 특히 비누공부분에서는 유의적으로 증가되어 있었다. Glutamine의 효과는 glycine 투여 효과와 비슷하거나 오히려 떨어지는 것으로 나타났다.

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

  • 진우정
    • Maxillofacial Plastic and Reconstructive Surgery
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    • 제15권1호
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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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인삼에서 추출한 글리코시드가 배양한 마우스 섬유아세포의 핵산합성에 미치는 영향 (Antiproliferative Effects of Panu ginseng Glycosides on DNA Synthesis in Cultured Mouse Fibroblasts)

  • Byun, Boo-Hyeong;Shin, In-Cheol;Lee, Lil-Ha;Joe, Cheol-O
    • Journal of Ginseng Research
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    • 제19권2호
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    • pp.114-116
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    • 1995
  • Panax ginseng ginsenosides were examined for their affects on the DNA synthesis. The DNA 1 synthesis was measured by the thymidine incorporation into NIH3T3 cells. The ginsenoside, panaxytriol, $Rh_1$ and $Rh_2$ showed reduced [$^{3}H$]-thymidine incorporation. However, other ginsenosides of $Rh_1$, $Rh_2$ and $Rh_3$ did not inhibit DNA synthesis. Among the various ginsenosides, ginsenoside $Rh_2$ was found to be the most inhibitory on DNA synthesis. We suggest $Rh_2$ as one of the potential choice of antiproliferative drugs.

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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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    • 제2권4호
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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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자궁내막증 환자의 복강액내 IGF가 자궁내막 기질세포 증식에 미치는 영향 (Effects of Insulin-like Growth Factor in Peritoneal Fluid of Patients with Endometriosis on the Proliferation of Endometrial Stromal Cells)

  • 김정구;서창석;김석현;최영민;문신용;이진용
    • Clinical and Experimental Reproductive Medicine
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    • 제26권3호
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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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Effect of Pyridoxine on Rifampicin Toxicity

  • Yun, Yeo-Pyo;Kim, Koan-Hoi;Kim, Hack-Seang;Chung, Jin-Ho
    • Archives of Pharmacal Research
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    • 제14권1호
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    • pp.73-77
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    • 1991
  • The effects of pyridoxine (PN) on rifampicin (RMF) toxicity were investigated by both in vivo and in vitro methods. RMF (30 mg/kg) was injected intraperitoneally and PN(150 mg/kg) was administered orally to rats for 10 consecutive days. After treatment, clinical chemistry and hematologic profiles were measured. RMF and PN plus RMF did not show any adverse effects at this in vivo experimental condition. Thymidine incorporations of mice bone marrow cells were examined in vitro. RMF showed a decrease in thymidine uptake in a dose-dependent manner, but PN showed a reversal of the thymidine uptake suppression caused by RMF (p<0.01). On the other hand, PN showed a decrease in thymidine uptake at a high concentration level.

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Toxoplasma gondii의 활성화된 uracil 도입 과정에 미치는 pyrimidine 대사 억제제의 영향 (Effects of pyrimidine salvage inhibitors on uracil incorporation of Toxoplasma gondii)

  • 윤지혜;남호우
    • Parasites, Hosts and Diseases
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    • 제28권2호
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    • pp.79-84
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    • 1990
  • Pyrimidine salvage과정 및 de novo 합성과정에 작용하는 억제제들이 Toxeplasma gondii의 특이한 uracil 도입 과정에 미치는 영향을 방사능을 표지한 uracil과 thymidine을 사용하여 검토하였다. Dihydrofolate reductase (DHFR)에 작용하는 억제제인 methotreBate, pyrimethamine 그리고 thymidylate synthase(75)의 경쟁적 억제제인 fluoro-uridine, fluoro-dUMP, fluoro-uracil을 각각 처리한 경우, 그 농도에 비례적으로 uracil 표지가 감소 하였다. OMP decarboxylase에 작용하는 억제제인 azauridine에 대해서는 100$\mu$M 농도 이상에서도 uracil 표지량에 변화가 거의 없었다. 본 결과로부터 Toxoplasma가 uracil을 DNA로 도입할 때 dihydrofolate reductase와 thymidylate synthase가 관여하는 TMP biosynthesis 과정이 중요함을 알 수 있었고, uracil salvage 과정이 있는 다른 기생성 원충의 경우와 비교했을 때 Toxoplasmn에 있어서도 효율적인 다기능의 DHFR-75 system의 존재를 예상할 수 있었다. Thymidine 도입의 양상은 모든 경우에 있어 HL-60세포만의 경우와 Toxoplasma가 함께 배양된 HL-60세포에서 차이가 없었다. 이로부터 thymidine은 Toxoplasmn 성장을 반영하지 않음을 알 수 있었다. Toxoplasma의 특이한 uracil 표지와 Toxoplasma에 배타적인 thymidine 표지는 기생원충과 숙주세포의 성장, DNA합성 정도 및 이들 과정에 미치는 제제들의 영향을 쉽고 간단하게 연구할 수 있는 방법이 된다.

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Alkoxybenzylcyanoguanidine Analogs as a Novel Class of Inhibitors for Restenosis

  • Lee, Sun-Kyung;Yi, Kyu-Yang;Hwang, Sun-Kyung;Suh, Jee-Hee;Lee, Byung-Ho;Yoo, Sung-Eun
    • Bulletin of the Korean Chemical Society
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    • 제25권7호
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    • pp.1003-1008
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    • 2004
  • A novel class of alkoxybenzylcyanoguanidine analogs as the inhibitors of restenosis was discovered, which showed the inhibitory effects on angiotensin II-induced cell proliferation, determined by $[^3H]$thymidine incorporation method. The compound, N'-(4-nitrophenyl)guanidine analog 19, showed 62% inhibition of $[^3H]$thymidine incorporation at 1 ${\mu}M$ concentration. In addition, the compound 19 inhibited intimal thickening dose-dependently after balloon injury, which suggests the therapeutic potential for restenosis.