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- 저자 : 황병두(Byung Do Hwang) (검색결과 4 건)
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NIH3T3 세포에 주입된 Taxol-Like Protein-35 의 작용 및 세포독성
선우재근(Jae Gun Sunwoo),이진우(Jin Woo Lee),황병두(Byung Do Hwang) 대한산부인과학회 2001 Obstetrics & Gynecology Science Vol.44 No.4
N/A Objective : To know whether the effects of TALP-35 introduced into NIH3T3 cells by osmotic lysis of pinosome (OLP method) can associates with microtubule, increases microtubule polymerization in cells and induces cytotoxicity. Methods : By osmotic lysis of pinosome (OLP method), TALP-35 was added to NIH3T3 cell. thereafter, distribution of TALP-35, indirect immunofluorescence microscopy, analysis of cells were done. Results : When TALP-35 was added to NIH3T3 cell homogenate, tubulin was cosedimented with TALP-35 and presented in microtubule pellets. TALP-35 was associated with microtubules and increased microtubule polymerization in NIH3T3 cells. Internalization of TALP-35 caused cell death. Cytotoxicity was found to be caused by apoptosis and G2/M phase block. Conclusion : TALP-35 infused in cell made microtubule polymerization increasing, stable and formed microtubule bundle. The mechanism of TALP-35-dependent was presented in cytotoxicity in NIH3T3 cell.
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12-O-Tetradecanoylphorbol-13-Acetate에 의한 HL-60 세포 분화유도중 Vimentin 유전자 전사조절에 대한 AP-1의 역할
임규,김진희,권도원,김승민,이명선,윤경아,손미영,박종일,윤완희,황병두 忠南大學校 癌共同硏究所 1998 癌共同硏究所 硏究誌 Vol.2 No.1
Purpose: To gain insight on the role of AP-1 in transcriptional regulation of vimentin gene during differentiation of HL-60 cells by 12-O-tetradecanoylphorbol-13-acetate (TPA), the levels of vimentin mRNA and AP-1 have been investigated with Northern blot hybridization and DNA mobility shift assay. Materials and Methods: HL-60 cells were grown in RPMI 1640 medium supplemented with 10% heat-inactivated fetal calf serum and antibiotics in a humidified 5% CO_(2) at 37°C. Total RNA was prepared by a modification of the method of Karlinsey et al. Northern blot hybridization was performed by the method of Virca et al. EcoRI fragment of pVIM-GEM was used as probe for vimentin mRNA. DNA mobility shift assay was performed by the method of Lim et al. End labeled DNA probe(Upper strand, 5'-CGCTTGATGAGTCAGCCG- 3') for AP-1 binding activity was mixed with nuclear extracts in a 20 μl reaction volume containing 300 mM KC1, 60 mM HEPES, pH 7.9, 25 mM MgCl_(2), 1 mM EDTA, 1 mM DTT, 60% glycerol, and 2μg of poly[dI-dC]. Results: TPA increased vimentin mRNA levels, with maximal stimulation reached at 24 hr. The level of vimentin mRNA was induced in proportion to the concentration of TPA. TPA-induced vimentin mRNA was almost reduced by actinomycin-D pretreatment. TPA-induced stimulation of vimentin gene was completely reduced by staurosporin pretreatment. In DNA mobility shift assay, AP-1 newly appeared at 24 hr during TPA-induced differentiation and was almost not detected after the pretreatment of staurosporin. Conclusions: These results suggest that the induction of vimentin mRNA during TPA-dependent differentiation in HL-60 cells may be mediated by protein kinases C signal transduction and AP-1 is important to transcriptional regulation.
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12-O-Tetradecanoylphorbol-13-Acetate에 의한 HL-60 세포 분화유도중 Vimentin 유전자 전사조절에 대한 AP-1의 역할
임규,김진희,권도원,김승민,이명선,윤경아,손미영,박종일,윤완희,황병두 충남대학교 생물공학연구소 1999 생물공학연구지 Vol.7 No.-
Purpose: To gain insight on the role of AP-1 in transcriptional regulation of vimentin gene during differentiation of HL-60 cells by 12-O-tetradecanoylphorbol-13-acetate (TPA), the levels of vimentin mRNA and AP-1 have been investigated with Northern blot hybridization and DNA mobility shift assay. Materials and Methods: HL-60 cells were grown in RPMI 1640 medium supplemented with 10% heat-inactivated fetal calf serum and antibiotics in a humidified 5% CO_2 at 37℃. Total RNA was prepared by a modification of the method of Karlinsey et al. Northern blot hybridization was performed by the method of Virca et al. EcoRI fragment of pVIM-GEM was used as probe for vimentin mRNA. DNA mobility shift assay was performed by the method of Lim et al. End labeled DNA probe(Upper strand, 5'-CGCTTGATGAGTCAGCCG- 3') for AP-1 binding activity was mixed with nuclear extracts in a 20 μl reaction volume containing 300 mM KCI, 60 mM HEPES, pH 7.9, 25 mM MgCI_2, 1 mM EDTA, 1 mM DTT, 60% glycerol, and 2 ㎍ of poly[dI-dC]. Results: TPA increased vimentin mRNA levels, with maximal stimulation reached at 24 hr. The level of vimentin mRNA was induced in proportion to the concentration of TPA. TPA-induced vimentin mRNA was almost reduced by actinomycin-D pretreatment. TPA-induced stimulation of vimentin gene was completely reduced by staurosporin pretreatment. In DNA mobility shift assay, AP-1 newly appeared at 24 hr during TPA-induced differentiation and was almost not detected after the pretreatment of staurosporin. Conclusions: These results suggest that the induction of vimentin mRNA during TPA-dependent differentiation in HL-60 cells may be mediated by protein kinases C signal transduction and AP-1 is important to transcriptional regulation.
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All-Trans Retinoic Acid에 의한 HL-60 세포 분화유도중 Vimentin 유전자 전사억제
김승민,박종일,윤완희,손미영,윤경아,임규,권도원,이명선,황병두 대한내분비학회 1998 Endocrinology and metabolism Vol.13 No.4
Background: Vimentin is the major intermediate-size filament in the cytoplasm of cells from mesenchymal origin. The HL-60 cell is a unique human leukemic cell line capable of terminal differentiation with several chemical inducers, and then the cell line becomes a fre#quently described model system for cell differentiation in vitro. Vimentin mRNA is reduced during all-trans retinoic acid (retinoic acid) -dependent differentication but increased by 12-0-tetradecanoylphorbol-13-acetate (TPA). In this paper, we have investigated on the mechanism of transcriptional repression of vimentin gene during retinoic acid-dependent differentication of HL-60 cell. Methods: HL-60 cells were grown in RPMI 1640 medium supplemented with 10% heat-inactivated fetal bovine serum and antibiotics in a humidified 5% CO at 37C. Total RNA was prepared by a modification of the method of Karlinsey et al. Northern blot hybridization was performed by the method of Virca et al. EcoRI fragment of pVIM-GEM was used as probe for vimentin mRNA. DNA mobility shift assay was performed by the method of Lim et al. End labeled DNA probe (Upper strand, 5-CGCITGATGAGTCAGCCG-3) for AP-1 binding activity was mixed with nuclear extracts in a 20 pL reaction volume containing 300 mM KCI, 60 mM HEPES, pH 7.9, 25mM MgC1, 1mM EDTA, 1mM DTT, 60% glycerol, and 2 pg of poly[dI-dC]. Results: The level of vimentin mRNA was decreased at 12 hours after retinoic acid treatment, and not detected at 48 hours. The level of vimentin mRNA was reduced in proportion to concentration of retinoic acid, Retinoic acid-reduced vimentin mRNA was no change in cells treated with cycloheximide. Retinoic acid-dependent decrease of vimentin mRNA was partially recovered by staurosporin pretreatment. In DNA mobility shift assay, AP-1 binding activity was reduced at 48 hr during retinoic acid-induced differentiation. Conclusion: These results suggest that the transcriptional repression of vimentin gene during retinoic acid-induced differentiation in HL-60 cells is correlated with reduction of DNA binding activity of AP-1 (J Kor Soc Endocrinol 13:601-611, 1998).
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