• Molecules and Cells
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• v.28 no.1
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• pp.31-36
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• 2009

Centrobin/Nip2 was initially identified as a centrosome protein that is critical for centrosome duplication and spindle assembly. In the present study, we determined the expression and subcellular localization of centrobin in selected mouse tissues. Immunoblot analysis revealed that the centrobin-specific band of 100 kDa was detected in all tissues tested but most abundantly in the thymus, spleen and testis. In the testis, centrobin was localized at the centrosomes of spermatocytes and early round spermatids, but no specific signal was detected in late round spermatids and elongated spermatids. Our results also revealed that the centrosome duplication occurs at interphase of the second meiotic division of the mouse male germ cells. The centrobin protein was more abundant in the mitotically active ovarian follicular cells and thymic cortex cells than in non-proliferating corpus luteal cells and thymic medullary cells. The expression pattern of centrobin suggests that the biological functions of centrobin are related to cell proliferation. Consistent with the proposal, we observed reduction of the centrobin levels when NIH3T3 became quiescent in the serum-starved culture conditions. However, a residual amount of centrobin was also detected at the centrosomes of the resting cells, suggesting its role for maintaining integrity of the centrosome, especially of the daughter centriole in the cells.

• Journal of Periodontal and Implant Science
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• v.27 no.3
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• pp.515-524
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• 1997

헤민과 메나디온 제한에 의한 Porphyromonas gingivalis(P. gingivalis) W50의 병독력의 변화를 검색하고자, 실험관내 병독력을 NIH 3T3 세포의 세포활성 변화로 관찰하였고, 생체내 병독력은 배양상청액을 ICR mouse 피하조직에 주사한 후의 염증반응을 관찰하였다. 헤민 존재 하에 배양한 P. gingivalis W50 배양상청액에 의한 mouse 3T3 세포의 세포활성은 헤민과 메나디온 없이 배양한 세포의 활성보다 낮았다. 헤민과 메나디온을 첨가하지 않고 배양한 세균의 생체내 병독력은 중등도의 염증세포 침윤과 울혈에 의한 출혈, 미약한 세포간질의 부종과 근육 파괴를 보였다. 메나디온 존재 하에서 배양한 세균은 미약한 염증세포의 침윤, 울혈에 의한 출혈 및 근육의 파괴가 관찰되었다. 헤민 존재하에서 중등도의 울혈에 의한 출혈, 미약한 세포간질의 부종, 염증세포의 침윤 및 근육파괴가 관찰되었다. 헤민과 메나디온 존재 하에서 배양한 세균은 심한 염증세포의 침윤과 중등도의 세포간질의 부종 및 울혈에 의한 출혈을 보였다. 이상의 연구 결과 P. gingivalis W50 배양 상층액의 병독력은 헤민에 의하여 영향을 받는 것으로 생각된다.

• Journal of Ginseng Research
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• v.19 no.2
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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.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.6
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• pp.856-865
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• 2007

Caveolin-1 of the caveolin family of proteins regulates mammary gland development and has been shown to play a contradictory role in breast tumor progression. A specific anti-Caveolin-1 antibody will be useful for functional study of Caveolin-1 in different tissues. In this study, we generated a rabbit polyclonal antibody that specifically recognizes the N-terminal amino acids 50-65 of Caveolin-1. This polyclonal antibody specifically reacted with Caveolin-1 extracted from cells of different species, including human epithelial A431 cells, goat primary mammary epithelial cells and mice fibroblast NIH 3T3 cells, by Western blotting. Endogenous Caveolin-1 protein expressing in cells and normal human tissues was detected by this polyclonal antibody using immunocytofluorescent and immunohistochemical staining, respectively. Furthermore, an apparent decrease in Caveolin-1 expression in tumorous breast and colon tissues was detected by this polyclonal antibody. In conclusion, we have identified amino acids 50-65 of Caveolin-1, which contains an epitope that is specific to Caveolin-1 and is conserved in the human, goat and mouse. In future, this anti-Caveolin-1 antibody can be used to examine the progression of breast and colon cancers and to study functions of Caveolin-1 in human, goat and mouse cells.

• Bulletin of the Korean Chemical Society
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• v.35 no.4
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• pp.1036-1042
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• 2014

In this study, we introduced the RRLR oligopeptide sequences on the surface of polyamidoamine (PAMAM) dendrimer and characterized the physical properties and gene carrier activity of the novel polymer using HEK 293, NIH3T3, and HeLa cells. The RRLR peptide sequences were derived from a mouse fibroblast growth factor 3 (FGF3) protein containing a bipartite NLS motif. The entire sequence of FGF3 is RLRRDAGGRGGVYEHLGGAPRRRK and it has two functional sequences RLRR and RRRK at N-terminus and C-terminus, respectively. In particular, PAMAM G4-RRLR conferred enhanced transfection efficiency and lower cytotoxicity compared with those of PEI 25 kDa, PAMAM G4-R, and PAMAM G4 in various cell lines. These results suggest that the introduction of N-terminal oligopeptides of FGF3 on the surface of PAMAM holds promise as an effective non-viral gene delivery carrier for gene therapy.

• Journal of Microbiology and Biotechnology
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• v.25 no.12
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• pp.1989-1996
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• 2015

Ice-binding proteins (IBPs) can inhibit ice recrystallization (IR), a major cause of cell death during cryopreservation. IBPs are hypothesized to improve cell viability after cryopreservation by alleviating the cryoinjury caused by IR. In our previous studies, we showed that supplementation of the freezing medium with the recombinant IBP of the Arctic yeast Glaciozyma sp. (designated as LeIBP) could reduce post-thaw hemolysis of human red blood cells and increase the survival of cryopreserved diatoms. Here, we showed that LeIBP could improve the viability of cryopreserved mammalian cells. Human cervical cancer cells (HeLa), mouse fibroblasts (NIH/3T3), human preosteoblasts (MC3T3-E1), Chinese hamster ovary cells (CHO-K1), and human keratinocytes (HaCaT) were evaluated. These mammalian cells were frozen in dimethyl sulfoxide (DMSO)/fetal bovine serum (FBS) solution with or without 0.1 mg/ml LeIBP at a cooling rate of -1℃/min in a -80℃ freezer overnight. The minimum effective concentration (0.1 mg/ml) of LeIBP was determined, based on the viability of HeLa cells after treatment with LeIBP during cryopreservation and the IR inhibition assay results. The post-thaw viability of mammalian cells was examined. In all cases, cell viability was significantly enhanced by more than 10% by LeIBP supplementation in 5% DMSO/5% FBS: viability increased by 20% for HeLa cells, 28% for NIH/3T3 cells, 21% for MC3T3-E1, 10% for CHO-K1, and 20% for HaCaT. Furthermore, addition of LeIBP reduced the concentrations of toxic DMSO and FBS down to 5%. Therefore, we demonstrated that LeIBP can increase the viability of cryopreserved mammalian cells by inhibiting IR.

• Macromolecular Research
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• v.10 no.3
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• pp.150-157
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• 2002

Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) is a microbial storage polymer with biodegradable properties. In order to improve the cell compatibility of PHBV surfaces, the physicochemical treatments have been demonstrated. In this study, physical method was corona discharge treatment and chemical method was chloric acid mixture solution treatment. The physicochemically treated PHBV film surfaces were characterized by the measurement of water contact angle, electron spectroscopy for chemical analysis, and scanning electron microscopy (SEM). The water contact angle of the physicochemically treated PHBV surfaces decreased from 75 to 30～40 degree, increased hydrophilicity. due to the introduction of oxygen-based functional group onto the PHBV backbone chain. The mouse NIH/3T3 fibroblasts cultured onto the physicochemically treated PHBV film surfaces with different wettability. The effect of the PHBV surface with different wettability was determined by SEM as counts of cell number and [$^3$H]thymidine incorporation as measures of cell proliferation. As the surface wettability increased, the number of the cell adhered and proliferated on the surface was increased. The result seems closely related with the serum protein adsorption on the physicochemically treated PHBV surface. In conclusion, this study demonstrated that the surface wettabilily of biodegradable polymer as the PHBV plays an important role for cell adhesion and proliferation behavior for biomedical application.

• Journal of Life Science
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• v.20 no.4
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• pp.623-631
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• 2010

Hericium erinaceus, an edible and medicinal mushroom belonging to the Basidiomycota family, has been used for curing gastric ulcers and stomach cancers in human beings and is also known to have good inhibitory effects on sarcoma 180 and Ehrlich carcinoma in mice. Neutral saline soluble (0.9% NaCl), hot water soluble and methanol soluble substances (hereinafter referred to as Fr. NaCl, Fr. HW and Fr. MeOH, respectively) were extracted from the fruiting body of the mushroom. In in vitro cytotoxicity tests, crude polysaccharides were not cytotoxic against cell lines such as Sarcoma 180, HepG2, HT-29 and NIH3T3 at concentrations of $10{\sim}2,000\;{\mu}g/ml$. Intraperitoneal injection with crude polysaccharides exhibited a life prolongation effect of 29.1~54.1% in mice previously inoculated with Sarcoma 180. Fr. Na increased the numbers of spleen cells by 2.9 fold at a concentration of $50\;{\mu}g/ml$ compared with the control. Fr. Na improved the immuno-potentiating activity of B lymphocytes by increasing alkaline phosphatase activity by 5.5 fold compared with the control at a concentration of $200\;{\mu}g/ml$. Fr. NaCl increased the numbers of peritoneal exudate cells and circulating leukocytes by 4 and 2.3 folds at a concentration of 50 mg/kg, respectively. Therefore, the crude polysaccharides extracted from the fruiting body of H. erinaceus could improve antitumor activities in mice.

• Journal of Life Science
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• v.18 no.10
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• pp.1348-1354
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• 2008

mi transcription factor (MITF) is important in regulating the differentiation of mast cells. In particular, MITF regulates the transcription of the mouse mast cell-specific serine protease (mMCP)-6 gene, which is generally expressed by the connective tissue-type of mast cells. In this study, we investigated alternative isoforms of MITF that regulate transcription of the mMCP-6 gene in bone marrow-derived cultured mast cells in mice. The expression of MITF isoforms was examined by RT-PCR. We observed that MITF-A, -E, -H and -Mc were expressed by mucosal-type mast cells cultured in the presence of IL-3, whereas the connective tissue-type mast cells cultured in the presence of stem cell factor (SCF) expressed MITF-A. Overexpression of MITF isoforms increased luciferase activity through the mMCP-6 promoter in NIH-3T3 cells and elevated the level of mMCP-6 expression in the MC/9 mast cell line. Moreover, mMCP-6 expression in mast cells was significantly inhibited by the depletion of MITF. The transcriptional activity and DNA binding of MITF-A was comparable to that of MITF isoforms, including MITF-E, -H, and -Mc. Our results therefore suggest that MITF-A may be an important isoform of MITF in regulating the transcription of mMCP-6 in mouse connective tissue mast cells.

• Korean Journal of Environmental Agriculture
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• v.16 no.2
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• pp.149-155
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• 1997

This study was carried out to investigate the effects of phenobarbital sodium(PB) and 3-methylcholanthrene(3-MC) on carbofuran cytotoxicity and to develop antitoxic agents based on the effectivness. Experimental groups for carbofuran cytotoxicity were divided into five groups ; medium alone and four treatments of carbofuran (1, 25, 50 and $100{\mu}M)$, and those for compensation effects were divided into six groups ; medium alone, $IC_{50}$ carbofuran and four combinations of carbofuran and PB or 3-MC($IC_{50}$ carbofuran plus 1, 25, 50, $100{\mu}M$ of PB and 3-MC, respectively). After incubation for 48 hrs under the same conditions, MTT(Tetrazolium MTT), NR(Neutral red) and SRB(Sulforhodamine B protein) assay were performed. Fifty percentage inhibition of MTT, NR, and SRB against carbofuran in rat fibroblast cell were 60.7, 82.5 and $87.0{\mu}M$, respectively. At the combination treatments of $IC_{50}$ of carbofuran and $100{\mu}M$ of PB, the significant compensation effects were observed from the results of MTT and NR but not from that of SRB absorbance. And at the combination treatments of $IC_{50}$ of carbofuran and 3-MC, the relatively significant compensation effects were found at $50{\mu}M$ 3-MC from the results of MTT and at $100{\mu}M$ 3-MC from that of NR and SRB absorbances, respectively. From the results of light microscopy, combination treatments of $carbofuran(IC_{50})$ and PB or 3-MC showed good regeneration in carbofuran toxicity of rat fibroblast cells. These results suggest that PB or 3-MC can compensate the cytotoxity of carbofuran insecticide in rat NIH3T3 fibroblast cells.