• Title/Summary/Keyword: NIH 3T3

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Cell proliferation effect of brown marine algae extracts on Mouse Fibroblast (해조류 추출물이 섬유아세포의 증식에 미치는 영향)

  • Ko, Ju-Young;Lee, Ji-Hyeok;Kim, Hyun-Soo;Kim, Hyung-Ho;Jeon, You-Jin
    • Journal of Marine Bioscience and Biotechnology
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    • v.7 no.1
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    • pp.28-34
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    • 2015
  • We examined cell regeneration efficiency of brown marine algae living in Jeju coast for search of a novel therapeutic device with cutaneous wound healing materials. The five algae were collected and compared with epidermal growth factor (EGF) as a positive control in the assays of cell proliferation and cell migration of NIH3T3 fibroblast cells. Among the 80% methanol extracts of these brown algae, the two algal extracts from Ishige foliacea and Colpomenia bullosa showed the proliferative effects of the cells similar to the effect of EGF. Besides it was found that Colpomenia bullosa extract significantly enhanced cell migration of NIH3T3 cell. In the study, therefore, we confirmed that the Colpomenia bullosa extract improved proliferation of NIH3T3 cell and a potential candidate for cultaneous wound healing.

Downregulation of bcl-xL Is Relevant to UV-induced Apoptosis in Fibroblasts

  • Nakagawa, Yuki;Okada, Seiji;Hatano, Masahiko;Ebara, Masaaki;Saisho, Hiromitsu;Tokuhisa, Takeshi
    • BMB Reports
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    • v.35 no.5
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    • pp.452-458
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    • 2002
  • Exposure to ultraviolet light (UV) induces apoptosis in mammalian cells, The caspase group of proteases is required for the appotosis. This pathway is initiated by a release of cytochrome c from the mitochondria into the cytosol. Several Bcl-2 family proteins can regulate the release of cytochrome c by stabilizing the mitochondrial membrane. Here we show that expression of the endogenous bcl-xL was strongly downregulated in NIH3T3 cells within 2 h after UV-C irradiation, and that of bax was upregulated from 8 h after irradiation. Apoptosis was induced in more than 50% of the NIH3T3 cells 48 h after irradiation. Constitutive overexpression of bcl-xL in NIH3T3 cells protected the UV-induced apoptosis by preventing the loss of mitochondrial membrane potential and the activation of caspase 9. There results suggest that downregulation of Bcl-xL is relevant to UV-induced apoptosis of tibroblasts.

Effect of Rhus verniciflua Stokes Extract and Fisetin on UVB-induced Apoptosis in NIH3T3 Cells (NIH3T3 세포에서 UVB에 의한 세포고사에 미치는 옻 추출물과 fisetin의 효과)

  • Kim Don Young;Hwang Eun Hee;Park Jong Kun
    • Journal of Life Science
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    • v.15 no.1 s.68
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    • pp.141-146
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    • 2005
  • For a long time Rhus vemiciflua Stokes (RVS) has been traditionally used as a herbal plant in Asia. In this study, we have investigated the effect of acetone extract of Rhus verniciflua Stokes (RVSE) and fisetin, a component of RVSE, on DNA damage response in NIH3T3 cells. Exposure of cells to DVB light $(200 J/m^2)$ and postincubation in growth medium for 48 hr resulted in a decrease of cell viability to about $10-20\%$ of nontreated control. Addition of various concentrations of RVSE in the postincubation medium, however, significantly increased the cell viability as compared with the values expected. The genotoxicity-decreasing effect was also demonstrated in cells exposed to UVB light and incubated in medium containing fisetin. The genotoxicity-decreasing effect of RVSE and fisetin was further demonstrated by various analyses including cell morphology studies, trypan blue exclusion assay and DAPI staining. By Annexin V binding analysis, RVSE and fisetin were shown to decrease the early apoptosis induced by UVB exposure. These results suggest the RVSE contain components that either increase the DNA repair or decrease the apoptosis in UVB-exposed cells.

Inhibitory Effects of Leachianone A from Sophora fIavescens Ait. against Cadmium Induced Cytotoxicity (고삼으로부터 분리된 Leachianone A의 카드뮴에 대한 독성억제 효과)

  • Young, Hee-Tae;Choi, Hwa-Jung;Baek, Seung-Hwa
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.22 no.5
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    • pp.1163-1167
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    • 2008
  • In this study, cytotoxicity of cadmium on NIH 3T fibroblasts was utilized in order to discover antitoxic compound in methanol extract of Sophora fIavascens Ait. There were treatment groups; control (medium only), $MTT_{50}$ group and five experimental groups. MTT {3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H- tetrazoliumbromide} assay was performed to evaluate the cytotoxicity of cell organelles and $IC_{50}$ was also measured. Accordingly we have examined the detoxification effects of methanol extract of S. flavescens Ait. and leachianone A (LA) on cadmium-treated NIH 3T3 fibroblasts ($IC_{50}=\;12.5{\mu}M$) to observe morphological changes by the light microscopy. Both S. flavescens Ait. methanol extract and LA showed inhibitory effects on cadmium-induced cytotoxicity. Furthermore, LA showed dose-dependency in detoxication. From these results, it is conceivable to suggest that LA from S. flavescens Ait. methanol extract is a potential antitoxic agent.

Specific Isoforms of Protein Kinase G Downregulate the Transcription of Cyclin D1 in NIH3T3

  • Lim, Seon Young;Soh, Jae-Won
    • Bulletin of the Korean Chemical Society
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    • v.34 no.4
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    • pp.1165-1169
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    • 2013
  • To elucidate the role of PKG isoforms in transcriptional control of cyclin D1, we employed a series of expression vectors of PKG $1{\alpha}$ and PKG $1{\beta}$ which encode HA-tagged wild type and constitutively active (SD and ${\Delta}N$) mutants. Our present study demonstrates that both the constitutively active mutants of PKG $1{\beta}$ downregulate the transcription of cyclin D1 when transiently transfected in NIH3T3 cells, whereas PKG $1{\alpha}$ mutants show weak inhibition. We further studied the transcriptional regulators of cyclin D1, such as, c-fos, NF-${\kappa}B$, and CRE by using the luciferase reporter assay. Constitutively active mutants of PKG $1{\beta}$ showed marked transcriptional downregulation of c-fos in NIH3T3 cells, whereas PKG $1{\alpha}$ downregulated c-fos to a lesser extent. We also found that the constitutively active mutants of PKG negatively regulated the activation of NF-${\kappa}B$ and CRE, suggesting their involvement in the regulation of cyclin D1.

Oligosaccharide-Linked Acyl Carrier Protein, a Novel Transmethylase Inhibitor, from Porcine Liver Inhibits Cell Growth

  • Seo, Dong-Wan;Kim, Yong-Kee;Cho, Eun-Jung;Han, Jeung-Whan;Lee, Hoi-Young;Hong, Sungyoul;Lee, Hyang-Woo
    • Archives of Pharmacal Research
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    • v.25 no.4
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    • pp.463-468
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    • 2002
  • We have previously reported on the identification of the endogenous transmethylation inhibitor oligosaccharide-linked acyl carrier protein (O-ACP), In this study, the role of the transmethylation reaction on cell cycle progression was evaluated using various transmethylase inhibitors, including O-ACP. O-ACP significantly inhibited the growth of various cancer cell lines, including NIH3T3, ras-transformed NIH3T3, MDA-MB-231, HT-1376, and AGS. In addition, exposure of ras-transformed NIH3T3 to O-ACP caused cell cycle arrest at the $G_0/G_1$ phase, which led to a decrease in cells at the S phase, as determined by flow cytometry. In contrast, transmethylase inhibitors did not affect the expression of $p21^{WAF1/Cip1}$, a well known inhibitor of cyclin dependent kinase, indicating that the cell cycle arrest by transmethylase inhibitors might be mediated by a $p21^{WAF1/Cip1}$-independent mechanism. Therefore, O-ACP, a novel transmethylase inhibitor, could be a useful tool for elucidating the novel role of methylation in cell proliferation and cell cycle progression.

Overexpression of Mouse Nck Transforms Mouse Febroblast NIH3T3

  • Kim, Young H.;Han, Sun-Mi;Kim, Moon G.;Park, Dong-Eun;Park, Sang D.;Seong, Rho H.
    • Animal cells and systems
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    • v.1 no.3
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    • pp.521-526
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    • 1997
  • We isolated a mouse nck cDNA from the thymus cDNA expression library. The cDNA encodes a 377 amino acid protein and displays 97% amino acid sequence identity to human oncogenic protein nck, which is composed almost exclusivelv of three src homology 3 (SH3) domains and one SH2 domain. The sequence analysis also showed that the isolated cDNA is the mouse counterpart of the human nck and different from the mouse grb4, which has been reported to be highly similar to the human nck and, therefore considered as a mouse nck, Northern blot analysis showed that the transcript of the gene was 1.8 kb and was highly expressed in the testis, thymus, and brain but moderately in the liver and lymph node. Western blot analysis showed that the size of the protein was about 47 kDa. Overexpression of the mouse Nck transformed a mouse fibroblast cell line, NIH3T3. The results clearly indicate that normal nck gene has transforming ability and provide an argument against a suggested possibility that the transforming ability of the human nck gene is due to a mutation(s) in the gene.

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Antioxidative Effect of Chelidonium majus Extract on Cultured NIH3T3 Fibroblasts Injured by Cadmium Chloride of Toxicant (독성물질인 염화카드뮴으로 손상된 배양 NIH3T3 섬유모세포에 대한 애기똥풀 추출물의 항산화 효과)

  • Kim, Tae-Yoon;Jekal, Seung-Joo
    • Korean Journal of Clinical Laboratory Science
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    • v.48 no.1
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    • pp.1-7
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    • 2016
  • The aim of this study was to evaluate the cytotoxicity of cadmium chloride ($CdCl_2$), toxicant, and the protective effect of Chelidonium majus (CM) extract on $CdCl_2$-induced cytotoxicity in cultured NIH3T3 fibroblasts. Cell viability, the effect of butylated hydroxytoluene (BHT) against $CdCl_2$, and the antioxidative effects including DPPH-free radical scavenging activity, superoxide anion-radical scavenging activity (SSA), and lactate dehydrogenase (LDH) activity were assessed. $CdCl_2$ caused a significant dose-dependent decrease in cell viability, and $XTT_{50}$ value was determined at 38.7uM of $CdCl_2$. It was determined as highly-toxic by Borenfreund and Puerner' toxic criteria. BHT of antioxidant significantly increased cell viability severely damaged by $CdCl_2$-induced cytotoxicity in these cultures. In the protective effect of CM extract on $CdCl_2$-induced cytotoxicity, CM extract significantly increased cell viability, DPPH-free radical scavenging activity, SSA and inhibitory activity of LDH. From these results, it is suggested that oxidative stress is involved in the cytotoxicity of $CdCl_2$, and CM extract showed protective efficacy on $CdCl_2$-induced cytotoxicity via antioxidative effects. Conclusively, natural resources like CM extract may be a putative antioxidative agent for the detoxification or diminution of toxicity correlated with oxidative stress.

The Melanin Inhibition, Anti-aging and Anti-inflammation Effects of Portulaca oleracea Extracts on Cells (쇠비름 추출물의 미백 및 항노화, 항염증 효과)

  • Zhang, Rui;Lee, Hyun-Jin;Yoon, Yeong-Min;Kim, Su-Mi;Kim, Hyun-Sook;Li, Shun Hua;An, Sung-Kwan
    • KSBB Journal
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    • v.24 no.4
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    • pp.397-402
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    • 2009
  • The Portulaca oleracea (P. oleracea) is a popular herbal medicine in East Asia that was known to possess detoxification, antifebrile and antifungal effects. In the present study, we examined the biological activities of ethanol extracts of P. oleracea under various conditions with NIH3T3, B16F10, and MCF-7 cell line model systems. Extracts of P. oleracea (0.5 mg/ml) showed inhibition of expression of tyrosinase, but does not suppress either of TYRP-1 or DCT expression on B16F10 cells. Extracts of P. oleracea (2 mg/ml) showed anti-inflammatory effects on TNF-$\alpha$-stimulated NIH3T3/$NF{\kappa}B$-Luc cells and increase of the synthesis of collagen on NIH3T3 (wild type) cells. These results suggest that extracts of P. oleracea could be used as a functional biomaterial in developing a skin whitening agent and having the anti-inflammatory, anti-wrinkle, and anti-aging activities.

Protective Effect of Agrimonia pilosa var. Extract on Cultured NIH3T3 Fibroblasts Damaged by Potassium Dichromate (크롬염으로 손상된 배양 NIH3T3 섬유모세포에 대한 짚신나물 추출물의 보호 효과)

  • Lee, Jun-Hee;Seo, Young Mi
    • Korean Journal of Clinical Laboratory Science
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    • v.51 no.2
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    • pp.205-213
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    • 2019
  • The protective effect of Agrimonia pilosa var. (AP) extract on potassium dichromate ($K_2Cr_2O_7$)-induced cytotoxicity in cultured NIH3T3 fibroblasts, was examined by performing an XTT assay for the cell viability and antioxidative effects, such as lactate dehydrogenase (LDH) activity and superoxide anion-radical (SAR) scavenging activity. In this study, $K_2Cr_2O_7$ decreased the cell viability significantly in a dose-dependent manner, and the $XTT_{50}$ value was determined to be $37.5{\mu}M$, which was highly-toxic according to the Borenfreund and Puerner' toxic criteria. The antioxidant, butylated hydroxytoluene (BHT), increased remarkably the cell viability damaged by $K_2Cr_2O_7$-induced cytotoxicity in these cultures. With regard to the protective effect of the AP extract on $K_2Cr_2O_7$-induced cytotoxicity, AP extract produced a significant increase in cell viability and antioxidative effects as the inhibitory ability LDH and SAR scavenging ability. These findings suggest that oxidative stress is involved in the cytotoxicity of $K_2Cr_2O_7$, and the AP extract effectively protected the cells from $K_2Cr_2O_7$-induced cytotoxicity by antioxidative effects. These results suggest that natural resources, such as AP extract, may be a putative therapeutic agent for the diminution or treatment of cytotoxicity induced by heavy metallic bases, such as $K_2Cr_2O_7$ correlated with oxidative stress.