• Title/Summary/Keyword: fibroblasts

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Gamma-Irradiation and Doxorubicin Treatment of Normal Human Cells Cause Cell Cycle Arrest Via Different Pathways

  • Lee, Seong Min;Youn, BuHyun;Kim, Cha Soon;Kim, Chong Soon;Kang, ChulHee;Kim, Joon
    • Molecules and Cells
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    • v.20 no.3
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    • pp.331-338
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    • 2005
  • Ionizing radiation and doxorubicin both produce oxidative damage and double-strand breaks in DNA. Double-strand breaks and oxidative damage are highly toxic and cause cell cycle arrest, provoking DNA repair and apoptosis in cancer cell lines. To investigate the response of normal human cells to agents causing oxidative damage, we monitored alterations in gene expression in F65 normal human fibroblasts. Treatment with ${\gamma}$-irradiation and doxorubicin altered the expression of 23 and 68 known genes, respectively, with no genes in common. Both agents altered the expression of genes involved in cell cycle arrest, and arrested the treated cells in $G_2M$ phase 12 h after treatment. 24 h after ${\gamma}$-irradiation, the percentage of $G_1$ cells increased, whereas after doxorubicin treatment the percentage of $G_2M$ cells remained constant for 24 h. Our results suggest that F65 cells respond differently to ${\gamma}$-irradiation- and doxorubicin-induced DNA damage, probably using entirely different biochemical pathways.

Identification of Inducible Genes during Mast Cell Differentiation

  • Lee Eunkyung;Kang Sang-gu;Chang Hyeun Wook
    • Archives of Pharmacal Research
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    • v.28 no.2
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    • pp.232-237
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    • 2005
  • Mast cells play an important role in allergic inflammation by releasing their bioactive mediators. The function of mast cells is enhanced by stimulation because of the induction of specific genes and their products. While many inducible genes have been elucidated, we speculated that a significant number of genes remain to be identified. Thus, we applied differential display (dd) PCR to establish a profile of the induced genes in bone marrow-derived mast cells (BMMCs) after they were co-cultured with 3T3 fibroblasts. To date, 150 cDNA fragments from the connective-type mast cells (CTMCs) were amplified. Among them, thirty cDNA fragments were reamplified for cloning and sequencing. The ddPCR strategy revealed that serine proteases were the most abundant genes among the sequenced clones induced during the maturation. Additionally, unknown genes from the co-culture of BMMCs with 3T3 fibroblasts were identified. We confirmed their induction in the CTMCs by Northern blot analysis and RT-PCR. Characterization of these induced genes during the maturation processes will provide insight into the functions of mast cells.

THE EFFECTS OF CHLORHEXIDINE ON THE ATTACHMENT AND GROWTH OF CULTURED HUMAN GINGIVAL FIBROBLASTS IN VITRO (인체 치은 섬유아세포의 성장과 부착에 관한 Chlorhexidine의 효과)

  • Lee, Ho;Lee, In-Kyu;Kim, Hyung-Seop
    • Journal of Periodontal and Implant Science
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    • v.23 no.3
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    • pp.507-516
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    • 1993
  • The Chlorhexidine(CHX) has been a widely used adjunt in periodontal therapy due to its bactericidal effect. In spite of the effects of CHX exhibits cytotoxic to human cells and delays granulation tissue formation. Therefore, understanding the effects of CHX on fibroblast attachment and cell growth will provide the rationale for its use during healing phase of periodontal surgery. This study was undertaken to examine the effects of standardized CHX-pretreated dentin slices and direct CHX exposure on human gingival fibroblasts. The results were as follow : 1. In experiment 1, there was a significant reduction in the number of fibroblast attachment in 0.12, 1%-pretreated groups relative to the control, 0.05%-pretreated groups(P<0.05). 2. In experiment 1, the control, 0.05%-pretreated groups showed considerable attachment and typical fibroblastic morphology, but 0.12, 1%-pretreated groups showed irregular, round-up (unattached) fibroblastic morphology. 3. In experiment 2, it appeared that all experimental groups exhibits significant inhibition of cell growth when compared with the control group.

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Force-mediated proinvasive matrix remodeling driven by tumor-associated mesenchymal stem-like cells in glioblastoma

  • Lim, Eun-Jung;Suh, Yongjoon;Kim, Seungmo;Kang, Seok-Gu;Lee, Su-Jae
    • BMB Reports
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    • v.51 no.4
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    • pp.182-187
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    • 2018
  • In carcinoma, cancer-associated fibroblasts participate in force-mediated extracellular matrix (ECM) remodeling, consequently leading to invasion of cancer cells. Likewise, the ECM remodeling actively occurs in glioblastoma (GBM) and the consequent microenvironmental stiffness is strongly linked to migration behavior of GBM cells. However, in GBM the stromal cells responsible for force-mediated ECM remodeling remain unidentified. We show that tumor-associated mesenchymal stem-like cells (tMSLCs) provide a proinvasive matrix condition in GBM by force-mediated ECM remodeling. Importantly, CCL2-mediated Janus kinase 1 (JAK1) activation increased phosphorylation of myosin light chain 2 in tMSLCs and led to collagen assembly and actomyosin contractility. Collectively, our findings implicate tMSLCs as stromal cells providing force-mediated proinvasive ECM remodeling in the GBM microenvironment, and reminiscent of fibroblasts in carcinoma.

The uptake of basic amino acids into fibroblasts was enhanced by PCA.

  • Ogasahara, Kazuko;Takino, Yoshinobu;Sakamoto, Kazutami
    • Proceedings of the SCSK Conference
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    • 2003.09b
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    • pp.145-148
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    • 2003
  • Previously, we reported that L-PCA enhanced blood circulation by modulating constitutive NO production. It was that L-PCA increased L-Arg uptake into endothelial cell, followed by the enhancement of NO production. Then we recommended the use of L-PCA for cosmetics, not only as humectants but also as enhancer of blood circulation. Since L-Arg is transported into endothelial cells by CAT (cationic amino acid transporter), it is expected that L-PCA also increase the uptake of basic amino acid, L-Lys. In this study, the uptakes of some amino acids into cells were evaluated by using 3H-labelled amino acid. Then we found the tendency that the uptake of L-Lys into endothelial cells was also enhanced by L-PCA. And the evident effect was observed in the epidermal fibroblasts, which had also CAT. Furthermore, it was found that the transportation of the other type of amino acids were not enhanced by L-PCA. That is to say, a famous moisturizer, L-PCA, has some effects on basic amino acid transport into cells.

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Metabolic Rebalancing of CR6 Interaction Factor 1-Deficient Mouse Embryonic Fibroblasts: A Mass Spectrometry-Based Metabolic Analysis

  • Tadi, Surendar;Kim, Soung Jung;Ryu, Min Jeong;Park, Taeseong;Jeong, Ji-Seon;Kim, Young Hwan;Kweon, Gi Ryang;Shong, Minho;Yim, Yong-Hyeon
    • Bulletin of the Korean Chemical Society
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    • v.34 no.1
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    • pp.35-41
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    • 2013
  • Metabolic analysis of CR6 interacting factor 1 (Crif1) deficient mouse embryonic fibroblasts with impaired oxidative phosphorylation has been carried out using LC-MS/MS and GC-MS methods. Metabolic profiles of the Crif1 deficient cells were comprehensively obtained for the first time. Loss of oxidative phosphorylation functions in mitochondria resulted in cancer-like metabolic reprogramming with consumption of majority of glucose carbon from up-regulated glycolysis to produce lactate, suppressed utilization of glucose carbon in the TCA cycle, increased amounts of amino acids. The changes in metabolic profile of the Crif1 deficient cells are most probably a consequence of metabolic reprogramming to meet the needs of energy balance and anabolic precursors in compensation for the loss of major oxidative phosphorylation functions.

Effect of Campsis grandiflora on Antioxidative Activity in UVB-irradiated Human Dermal Fibroblasts (사람 섬유아세포에서 UVB 조사에 대한 능소화 추출물의 항산화 효과)

  • Kim, Jin-Hwa;Lee, Bum-Chun;Zhang, Yong-He;Pyo, Hyeong-Bae
    • YAKHAK HOEJI
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    • v.49 no.2
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    • pp.174-179
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    • 2005
  • The human skin is constantly exposed to environmental irritants such as ultraviolet, smoke, chemicals. Free radicals and reactive oxygen species (ROS) caused by these environmen tal facts play critical roles in cellular damage. These irritants are in themselves damaging to the skin structure but they also participate the immensely complex inflammatory reaction. The purpose of this study was to investigate the skin cell protective effect of Campsis grandiflora extract on the UVB-irradiated human dermal fibroblasts (HDFs). We tested free radical and superoxide scavenging effect in vitro. C. grandiflora extracts had potent radical scavenging effect by 82% at $100{\mu}g/ml$, respectively. For testing intracellular ROS scavenging activity the cultured HDFs were analyzed by increase in DCF fluorescence upon exposure to UVB 20 $MJ/cm^2$ after treatment of C.grandiflora extracts. The results showed that oxidation of CM-DCFDA was inhibited by C.grandiflora extracts effectively and C.grandiflora extracts has a potent free radical scavenging activity in UVB- irradiated HDFs. In ROS imaging using confocal microscope we visualized DCF fluorescence in HDFs directly. In conclusion, our results suggest that C.grandiflora can be effectively used for the prevention of UV-induced adverse skin reactions such as radical production, and skin cell damage.

Necroptosis Is a Mechanism of Death in Mouse Induced Hepatocyte-Like Cells Reprogrammed from Mouse Embryonic Fibroblasts

  • Lee, Yun-Suk;Park, Kyung-Mee;Yu, Lina;Kwak, Ho-Hyun;Na, Hee-Jun;Kang, Kyung-Sun;Woo, Heung-Myong
    • Molecules and Cells
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    • v.41 no.7
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    • pp.639-645
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    • 2018
  • Liver transplantation is recommended for patients with liver failure, but liver donors are limited. This necessitates the development of artificial livers, and hepatocytes are necessary to develop such artificial livers. Although induced hepatocyte-like cells are used in artificial livers, the characteristics of mouse induced hepatocyte-like cells (miHeps) reprogrammed with embryonic fibroblasts have not yet been clarified. Therefore, this study investigated the mechanisms underlying the survival, function, and death of miHeps. miHeps showed decreased cell viability, increased cytotoxicity, decreased hepatic function, and albumin and urea secretion at passage 14. Addition of necrostatin-1 (NEC-1) to miHeps inhibited necrosome formation and reactive oxygen species generation and increased cell survival. However, NEC-1 did not affect the hepatic function of miHeps. These results provide a basis for development of artificial livers using hepatocytes.

Sulforaphane Inhibits Ultraviolet B-induced Matrix Metalloproteinase Expression in Human Dermal Fibroblasts

  • Lee, Sam Youn;Moon, Sun Rock
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.26 no.6
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    • pp.922-928
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    • 2012
  • Sulforaphane [1-isothiocyanato-4-(methylsulfinyl)-butane] is one of the most abundant isothiocyanates in some cruciferous vegetables, especially broccoli. Sulforaphaene has been shown to exhibit many pharmacological activities, including anti-oxidant, anti-inflammatory and anti-microbial activities. However, the anti-skin photoaging effects of sulforaphane have not yet been reported. In the present study, we investigated the inhibitory effects of sulforaphane on MMP-1 and -3 expressions of the human dermal fibroblasts via various in vitro experiments and elucidated the pathways of inhibition. Western blot analysis and real-time PCR revealed sulfiraphane inhibited UVB-induced MMP-1 and -3 expressions in a dose-dependent manner. UVB strongly activated nuclear factor-${\kappa}B$ (NF-${\kappa}B$) activity, which was determined by NF-${\kappa}B$ DNA binding activity. UVB-induced NF-${\kappa}B$ activation and MMP expression were completely blocked by sulforphane. These findings suggest that sulforaphane could prevent UVB-induced MMPs expressions through inhibition of NF-${\kappa}B$ activation.

Effects of Sipgeondaebo-tang Pharmacopuncture Extracts on the Collagenase Activity and Procollagen Synthesis in HS68 Human Fibroblasts and Tyrosinase Activity Original Articles (십전대보탕약침액(十全大補湯藥鍼液)의 사람 피부아세포의 콜라게나제 활성 및 프로콜라겐 합성과 티로시나제 활성에 미치는 영향)

  • Lee, Se-Na;Kim, Myung-Gyou;Kim, Myoung-Hee;Kim, Hyung-Jun;Jo, Hak-Jun;Leem, Kang-Hyun
    • Journal of Pharmacopuncture
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    • v.14 no.1
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    • pp.51-59
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    • 2011
  • Objectives : This study was designed to investigate the collagen metabolism and tyrosinase activity of Sipgeondaebo-tang Pharmacopuncture extracts (SP). Methods : The effect of SP on type I procollagen production and collagenase activity in human normal fibroblasts HS68 after UVB (312 nm) irradiation was measured by ELISA method. The tyrosinase activity after treatment of SP was measured as well. Results : Type I procollagen production was recovered by SP in UVB damaged HS68 cells. The increased collagenase activity after UVB damage was significantly recovered by SP. The tyrosinase activity was significantly reduced as well. However, the L-DOPA oxidation was not changed. Conclusion : SP showed the anti-wrinkle effects and whitening effects in vitro. These results suggest that SP may be a potential pharmacopuncture as an anti-aging pharmacopuncture treatments.