• Title/Summary/Keyword: NC-37 cell

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Effect of p16 on glucocorticoid response in a B-cell lymphoblast cell line

  • Kim, Sun-Young;Lee, Kyung-Yil;Jeong, Dae-Chul;Kim, Hak-Ki
    • Clinical and Experimental Pediatrics
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    • v.53 no.7
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    • pp.753-758
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    • 2010
  • Purpose: It has been suggested that p16 has a role in glucocorticoid (GC)-related apoptosis in leukemic cells, but the exact mechanisms have yet to be clarified. We evaluated the relationship between the GC response and p16 expression in a lymphoma cell line. Methods: We used p16 siRNA transfection to construct p16-inactivated cells by using the B-cell lymphoblast cell line NC-37. We compared glucocorticoid receptor (GR) expression, apoptosis, and cell viability between control (p16+NC-37) and p16 siRNA-transfected (p16-NC-37) cells after a single dose of dexamethasone (DX). Results: In both groups, there was a significant increase in cytoplasmic GR expression, which tended to be higher for p16+NC-37 cells than for p16- NC37 cells at all times, and the difference at 18 h was significant (P<0.05). Similar patterns of early apoptosis were observed in both groups, and late apoptosis occurred at higher levels at 18 h when the GR had already been downregulated ($P$<0.05). Cell viability decreased in both groups but the degree of reduction was more severe in p16+NC-37 cells after 18 h ($P$<0.05). Conclusion: These results suggest a relationship between GR expression and cell cycle inhibition, in which the absence of p16 leads to reduced cell sensitivity to DX.

Genomic changes of c-myc, c-H-ras in benzo(a)pyrene and dimethylbenz(a)anthracene treated human lymphoblast NC-37 cells (Benzo(a)pyrene과 dimethylbenz(a)anthracene에 의한 사람 림프아세포(NC-37)의 c-myc, c-H-ras 유전자 변화)

  • Cho, Moo Youn;Eo, Wan Kyu;Lee, Sang Uk;Jeong, In cheol
    • Journal of Life Science
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    • v.5 no.3
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    • pp.105-116
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    • 1995
  • To investigate genomic changes in c-myc gene by a chemical carcinogen, human lymphoblast NC-37 cells were exposed to benzo(a)pyrene(BP) and dimethylbenzanthracene(DMBA), and the c-myc gene expression was evaluated by Northern and Southern blot hybridization techniques. The results are as follows: When the genomic DNA of NC-37 cells exposed to several concentrations(1.25, 2.5 and 5ug/ml) of BP concentration. However, the c-myc gene was most significantly enhanced with 2.5ug/ml of BP. The expressions of c-myc gene in NC-37 cells was stimulated by BP and DMBA. Addition of TPA reduced the gene expression BP-treated cells, whereas it enhanced the gene expression in DMBA-treated cells. The expression of c-H-ras gene was slightly increased by treatment with BP and DMBA alone and in combination with TPA, however the magnitude of increase was not significantly different between each other. The expressions of c-myc c-H-ras genes in Burkitt's lymphoma cells were greater than those in NC-37 cells. When the DNA extracted from NC-37 cells exposed to various concentrations of BP were amplified by polymerase chain reaction using a primer set containing c-myc exon I, the amplified products were of the same size in all groups. To evaluate the BP toxicity in E.coli to which human c-myc gene-cloned pBR322 vector was inserted, Southern blot hybridization was conducted on c-myc genes digested with EcoRI/HindIII and Smal/Xbal restriction enzymes, and observing that in 2 ug/ml BP-treated cells a 3.5kb fragment was generated in addition to 1.3kb fragment which can be observed in normal cells. Direct nucleotide sequence analysis of polymerase chain reaction products showed a mutation of G$\longrightarrow$A transition at the Smal recognition site.

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A Study on the Skin Protection Effects of Cypress Essential Oil on the DNCB-induced Atopic Dermatitis in NC/Nga Mice (DNCB로 아토피피부염을 유발한 NC/Nga mice에서 사이프러스 에센셜 오일의 효능에 관한 연구)

  • Park, Chan-Ik
    • The Korea Journal of Herbology
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    • v.32 no.3
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    • pp.37-44
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    • 2017
  • Objectives : The application of DNCB (1-chloro-2,4-dinitrochlorobenzene) can cause cellular immunity allergic reaction such as erythema or edema on NC/Nga mice and combinational function of cells participating in immunity increase inflammatory mediator. In this study, the effects of cypress essential oil on NC/Nga mice have been assessed. Methods : Male SPF NC/Nga mice aged 8 weeks have been used for atopic dermatitis induction skin. 1% DNCB was applied on ears and backs of which hair was removed using clipper on $1^{st}$ day and 0.4% DNCB was applied three times a week for 3 weeks. In this study, cypress essential oil has been treated 1 time a day for 3 weeks after application of DNCB to induce atopic dermatitis on skin for further experiments. Results : This study shows that inhalation or application of cypress essential oil reduced edema in ears and the thickness of epidermis induced by DNCB treatment. And it can be known that treatment of cypress essential oil inhibited mast cell proliferation and reduced IgE level similar to that of the negative control especially when cypress essential oil was inhaled by the mice. Synthetic oil showed the effects lower than those of cypress essential oil. Conclusions : Inhalation or direct application of cypress essential oil on skin reduced IgE level in blood and prohibits the proliferation of mast cell, from which it can be known that cypress essential oil can be effectively used to reduce the symptoms of atopic dermatitis.

Differentiation Induction of Dendritic Cell Phenotypes from Human Leukemic Cell Lines

  • Lee, Dae-Heui;Park, Jae-Sun;Eo, Wan-Kyu;Kim, Woo-Mi;Kang, Koo-Il
    • The Korean Journal of Physiology and Pharmacology
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    • v.5 no.1
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    • pp.79-86
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    • 2001
  • Recent clinical studies have shown that a high proportion of patients with acute promyelocytic leukemia (APL) achieve complete remission after treatment with all-trans retinoic acid (ATRA). However, most patients who receive continuous treatment with ATRA relapse and develop ATRA-resistant leukemia. Dendritic cells (DCs) are important antigen-presenting cells in the development of antileukemic T-cell responses. In this study, we investigated the strategies to overcome ATRA resistance of APL cells by inducing the differentiation of DCs from human leukemic cell lines for the developtment of adoptive immunotherapy. CD83 was used as a mature DC marker in this study and the expression of CD83 mRNA was determined by RT-PCR method. The promyelocytic leukemic cell line HL-60, B lymphoblast cell lines RPMI 7666 and NC-37 could be induced to dendritic cells in vitro. Treatment of HL-60 with phorbol 12-myristate 13-acetate (PMA) resulted in the expression of myeloid-related DC phenotypes, while treatment of RPMI 7666 with fms-like tyrosine kinase 3 ligand (Flt3-ligand, FL) and treatment of NC-37 with PMA and FL led to the expression of lymphoid-related DC phenotypes. In conclusion, myeloid-related DC phenotypes and lymphoid-related DC phenotypes could be generated from HL-60, NC-37 and RPMI 7666 cell lines, respectively. These DC phenotypes can potentially be used to generate antileukemic T cells in vitro for adoptive immunotherapy.

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Preparation and Structure of $Re(≡NC_6H_5)(DPPE)CI_3$, $[DPPE=Ph_2PCH_2CH_2PPh_2]$ ($Re(≡NC_6H_5)(DPPE)CI_3$화합물의 합성 및 구조)

  • 박병규;정건수
    • Korean Journal of Crystallography
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    • v.6 no.2
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    • pp.93-97
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    • 1995
  • Re(≡NC6H5)(PPh3)2CI3, I, reacted with, 1,2-bis(diphenylphosphino)ethane (DPPE) to give fac-Re(≡NC6H5)(DPPE)CI3, II. The product has been characterized by 1H-NMR, elemental analysis, and X-ray crystallography. II Crystalizes in the monoclinic space group Pc, with cell parameters a=11.083(3)Å, b=10.930(1)Å, c=14.081(2)Å, β=108.37(2)°, Z=2. Least-squares refinement of the structure led to a R(wR2)factor of 0.0254(0.0607) for 2888 unique reflections of I>2σ(I) and for 352 variables.

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Studies on the Nuclei Adduction and Expression of c-myc Gene by Benzo(a)pyrene and Doxorubicin in Human NC-37 Cells (사람 NC-37 세포에서 Benzo(a)pyrene과 Doxorubicin에 의한 Nuclei내전과 c-myc 유전자의 발현에 대한 연구)

  • 김호찬;정인철;조무연
    • Journal of Life Science
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    • v.8 no.4
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    • pp.400-409
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    • 1998
  • Formation of adduct was studied in benzo(a)pyrene(BP)- and doxorubicin(Dx)-treated human NC-37 cells and isolated nuclei. Major adducts formed were determined by fluorescence absorption spectrophotometery and DNA-lin-ked protein assay. When isolated nuclei were exposed to carcinogens BP and DMBA, and anticancer drugs m-AMSA, ellipticine and Dx, varying degrees of adduct formation occured between DNA-protein complex and these drugs. When the mixture was centrifuged 1.7 M sucrose solution, binding BP and DMBA appeared to be similar between the sediment and the supernatant. When the sediment was centrifuged again with 0.35% polymin-P, the amount of BP bound was 2-fold greater in the protein(1077$\pm$55cpm) than in DNA fraction (470$\pm$20cpm), whereas that of DMBA was 1.6-fold greater in the DNA than in protein fraction. In the case of m-AMSA, ellipticine and Dx, the amount of binding was slightly greater in supernatant than in sediment in centrifugation with 1.7 M sucrose, and more than 3 times greater in the DNA- than in protein- fraction in centrifugation with 0.35% polymin P. DNA fractions which associated with a subset of nonhistone chromosomal protein were isolated from NC-37 cells exposed to $^{3}$H-BP and $^{14}$C-Dx. They were separated into two distince components DNA-S and DNA-P by centrifugation with 2M Nacl chromatin extraction. The results indicated that the amount of $^{3}$H-BP bound was 6.0-fold greater in DNA-P as compared with DNA-S, while that of $^{14}$C-Dx binding appreaed to be 6.2-fold greater in DNA-S than in DNA-P fraction. When $^{3}$H-BP binding wasdetermined in the presence of cold Dx, the amount of binding was reduced only in the DNA-P fraction, indicating that the interaction between DNA and protein is decreased. Gene expression by these drugs, BP treated cells were increased to compare with nomal cells but reduced by treatment with BP-Dx. These results suggest that the protein moiety which tightly bound to DNA-P fraction may play an important role in the regulation of gene expression.

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Effects of Atopy Cream-combined with Jawoongo Ointment on the Development of Atopic Dermatitis-like Skin Lesions in NC/Nga Mouse (아토피 피부염 동물 모델에서 아토피 크림과 자운고(紫雲膏)의 병용도포가 피부염에 미치는 영향)

  • Yeo, Eui-Ju;Han, Jae-Kyung;Kim, Yun-Hee
    • The Journal of Pediatrics of Korean Medicine
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    • v.23 no.1
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    • pp.37-72
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    • 2009
  • Objectives The purpose of this study is to investigate the suppressive effects of Atopy cream-combined with Jawoongo ointment (A-J), on the development of atopic dermatitis-like skinlesions in NC/Nga mouse. Methods We evaluated clinical skin score, hematology and Serum total IgE and IgG1 of NC/Nga atopic dermatitis mouse, analyzed the cytokine level, total cell number, Immunohistochemical staining and Histological features of axillary lymph node(ALN), draining lymph node(DLN), peripheral blood mononuclear cells(PBMCs) and dorsal skin tissue in NC/Nga mouse. Results A-J decreased the clinical skin score, total cell number of WBC, platelet, neutrophils, eosinophils in blood, Serum total IgE & IgG1, IL-5, IL-13. Also, total cell number of ALN and dorsal skin tissue, Absolute cell number of $CD3e^+$&$CD19^+$, $CD4^+$&$CD8^+$, $CD3^+/CCR3^+$, $CCR3^+$, $CD3^+/CD69^+$, $CD3^+/CXCR5^+$ in ALN, PBMCs, Absolute cell number of $CCR3^+$, $CD3^+/CD69^+$, $CD11b^+/Gr-1^+$ in dorsalskin tissue, Eotaxin2 mRNA, CCR3 mRNA in dorsal skin tissue and gene expression of IL-5 mRNA, IL-13 mRNA in ALN decreased significantly. Furthermore, thickness of epidermis, infiltrated inflammatory immune cell and mast cell in dermis, histologic infiltration of mast cell, the size of inflammatory lymphocytes cells and plasma cells in ALN and histologic infiltration of CD4+ & CCR3+ in ALN and dorsal skin tissue decreased significantly. However, total cell number of DLN, absolute cell number of $CD3e^+$&$CD19^+$, $CD4^+$&$CD8^+$, $B220^+/CD23^+$, $CD3^+/CD69^+$ increased significantly. Conclusions A-J was the successful treatment of atopic dermatitis in a NC/Nga mouse model.

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Effect of Polygonum multipolarum extract on the allergic reaction of NC/Nga mice causing atopic dermatitis (하수오 추출물(Polygonum multiflorum)이 아토피 피부염 유발 NC/Nga 생쥐의 알레르기 반응에 미치는 영향)

  • Moon, Ji-Sun;Choi, Eun-Young
    • Journal of the Korean Applied Science and Technology
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    • v.37 no.1
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    • pp.66-75
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    • 2020
  • Atopic dermatitis (AD) usually develops in patients with an individual or family history of allergic diseases, and is characterized by chronic relapsing inflammation seen specially in childhood, association with IgE hyperproduction and precipitation by environmental factors. and wished to examine closely effect that Polygonum multiflorum isolated PM-E and PM-70M orally adminstration used to atopy dermatitis disease patient get in atopy eruption control experimentally. Atopic dermatitis is a chronically relapsing inflammatory skin disease. Animal models induced by relevant allergens play a very important role in the elucidation of the disease. This study was investigated the anti-allergic effect of PM-E and PM-70M on BMAC induced atopic dermatitis in NC/Nga mice. We summerized as the follow. PM-E and PM-70M significantly reduced the skin number of total cell number, CD4+ and CD11b+/Gr-1 cell compared with positive control and decreased the invasion of CD4+ cell in dorsal skin tissue compared with positive control group by using immunohistochemical staining and chemokine such as eotaxin and CCR3 compared with positive control group. PM-E and PM-70M markedly suppressed invasion and edema of leukocytes and mast cell in dorsal skin. Taken together, these findings suggested that PM-E and PM-70M has an anti-allergic activity and this might be useful for the clinical application to treat allergic diseases such as atopic dermatitis.

Differential Effects of Green Tea Polyphenol in the ${\gamma}-irradiation$ Induced Human Leukemic and Lymphoblastic Cell Damage (녹차 폴리페놀이 감마선조사에 의한 백혈병과 림프구모세포의 손상에 미치는 영향의 차이)

  • Jeong, Hwan-Jeong;Kim, Eun-Mi;Min, Jung-Jun;Bom, Hee-Seung;Kim, Young-Ho;Jeong, Young-Do;Kim, Chang-Guhn
    • The Korean Journal of Nuclear Medicine
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    • v.37 no.5
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    • pp.308-316
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    • 2003
  • Purpose: The green tea polyphenol (GTPP) has been known to exert antioxidant activity as a radical scavenger as well as cancer preventive and cancer growth inhibition effect. The aim of this study was to identify whether GTPP not only potentiate the growth inhibition effect in ${\gamma}-irradiated$ human cancer cell but also exert protection action for irradiated human normal cell. Materials and Methods: GTPP (80% catechin including >45% EGCG) added in the HL60, human leukemia, and NC37, human lymphoblast, before irradiation. After establishing the amount of GTPP and the dose of radiation, the cells were treated with the GTPP for 6 hours and irradiated with the determined doses. Results: Viability when $10{\mu}g/ml$ GTPP added before ${\gamma}-irradiation$ with 1 Gy to NC37 cells was not different in comparison with control but it when was irradiated with 3 Gy significantly different (1 Gy;P=0.126, 3 Gy;P=0.010). $20{\mu}g/ml$ GTPP did not show significant difference in both NC37 cells irradiated with 1 Gy and 3 Gy (1 Gy;P=0.946, 3 Gy;P=0.096). Viabilities were significantly decreased with concentration of additional GTPP in HL60 with 1 or 3 Gy (1 Gy $69.0{\pm}1.7%\;vs\;42.4{\pm}1.3%,\;3\;Gy;\;66.9{\pm}3.9%\;vs\;44.2{\pm}1.6%$). Conclusion: In vitro study, we certified that when the cells were irradiated with dose below 3 Gy, GTPP provide not only anticancerous effect against cancer cells but also radioprotective effect in normal cells simultaneously. Theses results suggest the possibility that consumption of green tea could give the radioprotective effect and maximize the effect on internal radiation such as radioiodine therapy concomitantly.

Genes Associated with Radiation Adaptive Response Induced by Low Level Radiation from $^{99m}Tc$ in Human Cell Lines (인체세포주에서 저선량 $^{99m}Tc$에 의해 발현되는 방사선 적응반응에 관련된 유전자에 관한 연구)

  • Kwon, An-Sung;Bom, Hee-Seung;Choi, Chan;Kim, Ji-Yeul;Lim, Wook-Bin
    • The Korean Journal of Nuclear Medicine
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    • v.35 no.5
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    • pp.313-323
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    • 2001
  • Purpose: The purpose of this study was to search activated genes that could be related to radiation adaptive response (RAR) induced by low-level radiation from $^{99m}Tc$ in human cell lines. Methods: We used gene discovery array (GDA) and representational difference analysis (RDA) methods. $^{99m}Tc$-pertechnetate was added to $2{\times}106/mL$ NC-37 cells (human lymphoblastic cells) to make concentrations ranging from 148 MBq/mL to 148 Bq/mL by serial 10 fold dilutions. After 44 hours, 2 Gy gamma irradiation was given to them using a Cs-137 cell irradiator. Results: As compared to the control (Con) group to which no $^{99m}Tc$ was added, those cells to which 148 and 14.8 KBq of $^{99m}Tc$ were added showed significantly lower damage to chromosomes, which was evaluated by metaphase analysis. Cells with 148 KBq $^{99m}Tc$ (T148 group) showed most significant protection. Activated genes in the T148 group as compared to Con group were evaluated by GDA and GDA methods. GDA revealed genes of casein kinase 2 (CK2) beta chain, immunoglobulins (lg), human leukocyte antigen (HLA)-B, and two novel genes. Twenty RAR related clones were selected by RDA method. The size of those genes was from 234 to 603 base pairs. Conclusions: RAR was induced by low dose irradiation from $^{99m}Tc$ in NC-37 cell lines. Genes related to the response included CK2, lg, HLA-B in human lymphoblastic cell lines.

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