• 제목/요약/키워드: regulatory factor

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EFFECT OF PDGF AND $TGF-{\beta}1$ ON CELL ACTIVITY OF HUMAN GINGIVAL FIBROBLAST AND PERIODONTAL LIGAM ENT CELL IN VITRO (PDGF와 $TGF-{\beta}1$이 배양 인체 치은 섬유모세포와 치주인대세포의 활성에 미치는 영향)

  • Chung, Soon-Kyu;Nam, Goong-Hyuk;Shin, Hyung-Shik
    • Journal of Periodontal and Implant Science
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    • v.25 no.1
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    • pp.133-145
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    • 1995
  • The migration and proliferation of periodontal ligament cells are desired goal of periodontal regeneration therapy. PDGF and $TGF-{\beta}1$ are well known to regulate the cell activity of mesenchymal origin cell. The purpose of this study was to determine the effects of these growth factors on human gingival fibroblast and periodontal ligament cell actvity, and to identify the regulatory effect of $TGF-{\beta}1$ on the response to PDGF by MIT assay. Human gingival fibroblast and periodontal ligament cells were cultured from extracted teeth for non-periodontal reason. Cultured human gingival fibroblast and periodontal ligament cells in vitro were treated with polyperpetide growth factor PDGF and $TGF-{\beta}1$ in both a dose and time - dependent manner. Cell morphology were determined by inverted microscope and cell acitivity were determined by MIT assay. The result of this study demonstrated that PDGF and $TGF-{\beta}1$ were not changed the morphology of these cell compared with control group. PDGF or $TGF-{\beta}1$ increased cell activity of periodontal ligament cell in dose and time dependent manner but gingival fibroblast were decreased to the level of control group at third day. Additionally, incubation with $TGF-{\beta}1$ addition to PDGF resulted in a enhanced cell activity of PDGF. Therefore, cell acitivty of gingival fibroblast were not changed compared with control group. This stiudy demonstrates that PDGF and $TGF-{\beta}1$ are major mitogens for human periodontal ligament cell in vitro, and $TGF-{\beta}1$ is a regulator of cell activity to PDGF in human gingival fibroblast and periodontal ligament cell.

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Specific Knockdown of Nanog Expression by RNA Interference in P19 Embryonal Carcinoma Stem Cells (P19 배아 암종 줄기세포에서 RNA 간섭에 의한 Nanog 유전자 발현의 특이적 억제)

  • Choi, Seung-Cheol;Lim, Do-Sun
    • Development and Reproduction
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    • v.12 no.2
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    • pp.159-168
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    • 2008
  • Nanog is a newly identified member of the homeobox family of DNA binding transcription factors that functions to maintain the undifferentiated state of stem cells. However, molecular mechanisms underlying the function of Nanog remain largely unknown. To elucidate the regulatory roles of Nanog involved in maintenance of P19 embryonal carcinoma (EC) stem cells, we transfected three small interfering RNA (siRNA) duplexes targeted against different regions of the Nanog gene into P19 cells. The Nanog siRNA-100 duplexes effectively decreased the expression of Nanog up to 30.7% compared to other two Nanog siRNAs, the Nanog siRNA-400 (67.9 %) and -793 (53.0%). When examined by RT-PCR and real-time PCR, the expression of markers for pluripotency such as Fgf4, Oct3/4, Rex1, Sox1 and Yes was downregulated at 48 h after transfection with Nanog siRNA-100. Furthermore, expression of the ectodermal markers, Fgf5 and Isl1 was reduced by Nanog knockdown. By contrast, the expression of other markers for pluripotency such as Cripto, Sox2 and Zfp57 was not affected by Nanog knockdown at this time. On the other hand, the expression of Lif/Stat3 pathway molecules and of the endoderm markers including Dab2, Gata4, Gata6 and the germ cell nuclear factor was not changed by Nanog knockdown. The results of this study demonstrated that the knockdown of Nanog expression by RNA interference in P19 cells was sufficient to modulate the expression of pluripotent markers involved in the self-renewal of EC stem cells. These results provide the valuable information on potential downstream targets of Nanog and add to our understanding of the function of Nanog in P19 EC stem cells.

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Screening of Differentially Expressed Genes by Desferrioxamine or Ferric Ammonium Citrate Treatment in HepG2 Cells

  • Park, Jong-Hwan;Lee, Hyun-Young;Roh, Soon-Chang;Kim, Hae-Yeong;Yang, Young-Mok
    • BMB Reports
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    • v.33 no.5
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    • pp.396-401
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    • 2000
  • A differential display method is used to identify novel genes whose expression is affected by treatment with ferric ammonium citrate (FAC) or desferrioxamine (DFO), an iron chelating agent in the human hepatoblastoma cell line (HepG2). These chemicals are known to deplete or increase the intracellular concentration of iron, respectively. Initially, we isolated seventeen genes whose expressions are down- or up regulated by the treatment of the chemicals, as well as their four differentially expressed genes that are designated as clone-1, -2, -3, and -4. These are further characterized by cDNA sequencing and Northern blot analysis. Through the cDNA sequencing, as well as comparing them to genes published using the NCBI BLAST program, we identified the sequence of the clone-1 that is up-regulated by the treatment of DFO. It is identical to the human insulin-like growth factor binding protein-1 (IGFBP-1). This suggests that the IGFBP-1 gene in the HepG2 cell is up-regulated by an iron depletion condition. Also, the expression of the clone-3 and -4 is up-regulated by FAC treatment and their eDNA sequences are identical to the human ferritin-fight chain and human NADH-dehydrogenase, respectively. However, the sequence of the clone-2 has no significant homology to any other known gene. Therefore, we suggest that changes of the cellular iron level in the HepG2 cell affects the transcription of cellular genes. This includes human IGFBP-1, ferritin-fight chain, and NADH-dehydrogenase. Regulation of these gene expressions may have an important role in cellular functions that are related to cellular iron metabolism.

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Post-transcriptional Regulation of Gcn5, a Putative Regulator of Hox in Mouse Embryonic Fibroblast Cells

  • Lee, You-Ra;Oh, Ji-Hoon;Kong, Kyoung-Ah;Kim, Myoung-Hee
    • Biomedical Science Letters
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    • v.18 no.2
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    • pp.165-168
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    • 2012
  • Hox proteins containing DNA-binding homedomain act as transcription factors important for anteroposterior body patterning during vertebrate embryogenesis. However, the precise mechanisms by which signal pathways are transduced to regulate the Hox gene expression are not clear. In the course of an attempt to isolate an upstream regulatory factor(s) controlling Hox genes, protein kinase B alpha (Akt1) has been identified as a putative regulator of Hox genes through in silico analysis (GEO profile). In the Gene Expression Omnibus (GEO) dataset GDS1784 at the NCBI (National Center for Biotechnology Information) site, Hox genes were differentially expressed depending on the presence or absence of Akt1. Since it was not well known how Akt1 regulates the specific Hox genes, whose transcription was reported to be regulated by epigenetic modifications such as histone acetylation, methylation etc., the expression of Gcn5, a histone acetyltransferase (HAT), was analyzed in wild type (WT) as well as in $Akt1^{-/-}$ mouse embryonic fibroblast (MEF) cells. RT-PCR analysis revealed that the amount of Gcn5 mRNA was similar in both WT and $Akt1^{-/-}$ MEFs. However, the protein level of Gcn5 was significantly increased in $Akt1^{-/-}$ MEF cells. The half life of Gcn5 was 1 hour in wild type whereas 8 hours in $Akt1^{-/-}$ MEF. These data all together, indicate that Gcn5 is post-transcriptionally down-regulated and the protein stability is negatively regulated by Akt1 in MEF cells.

G1 Arrest of the Cell Cycle by Onchungeum in Human Hepatocarcinoma Cells (온청음(溫淸飮)이 인체 간암세포의 세포주기 G1 Arrest에 미치는 영향)

  • Goo, In-Moo;Shin, Heung-Mook
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.22 no.4
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    • pp.821-828
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    • 2008
  • Onchungeum, a herbal formula, which has been used for treatment of anemia due to bleeding, discharging blood and skin disease. In the present study, it was examined the effects of extract of Onchungeum (OCE) on the growth of human hepatocarcinoma cell lines Hep3B (p53 null type) and HepG2 (p53 wild type) in order to investigate the anti-proliferative mechanism by OCE. Treatment of Hep3B and HepG2 cells to OCE resulted in the growth inhibition in a dose-dependent manner, however Hep3B cell line exhibited a relatively strong anti-proliferative activity to OEC. Flow cytometric analysis revealed that OCE treatment in Hep3B cells caused G1 phase arrest of the cell cycle, which was associated with various morphological changes in a dose-dependent fashion. RT-PCR and immunoblotting data revealed that treatment of OCE caused the down-regulation of cyclin D1 expression, however the levels of cyclin E expression were not changed by OCE. The G1 arrest of the cell cycle was also associated with the induction of Cdk inhibitor p27 by OCE. Because the p53 gene is null in Hep3B cells, it is most likely that the induction of p21 is mediated through a p53-independent pathway. Moreover, p27 detected in anti-Cdk4 and anti-Cdk2 immunoprecipitates from the OCE-treated cells, suggesting that OCE-induced p27 protein blocks Cdk kinase activities by directing binding to the cyclin/Cdk complexes. Furthermore, OCE treatment potently suppresses the phosphorylation of retinoblastoma proteins and the levels of the transcription factor E2F-1 expression. Taken together, these results indicated that the growth inhibitory effect of OCE in Hep3B hepatoma cells was associated with the induction of G1 arrest of the cell cycle through regulation of several major growth regulatory gene products.

Study on Establishment of the Industrial Wastewater Effluent Limitations Based on Best Practicable Control Technology Currently Available - Case Study for the Pulp, Paper and Paper Board Manufacturing (실용 가능한 최적처리기술에 근거한 산업폐수 배출허용기준 설정 연구 - 펄프.종이 및 종이제품 제조시설 적용 사례)

  • Kim, Jaehoon;Shin, Jinsoo;Lee, Chulgu;Lee, Jungyoung;Lee, Youngsun;Yu, Soonju
    • Journal of Korean Society on Water Environment
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    • v.28 no.4
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    • pp.608-614
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    • 2012
  • The effluent limitation of industrial wastewater is based on uniform regulatory criteria for effluent discharge facilities of all in Korea. But, an individual effluent limitation on each effluent discharge facility is widely applicable for regulation of industrial wastewater in US.EPA. To decide an individual effluent limitation, TBEL (Technology-based effluent limitation) and WQBEL (Water quality-based effluent limitation) are used. TBEL is based on the capability of a treatment technology to reduce the pollutants. WQBEL is based on ambient water quality standards. In this study, TBEL were derived for the pulp, paper and paper board manufacturing based on best practicable control technology currently available. It was suggested that effluent limitations were $BOD_5$ 4.7 mg/L, $COD_{Mn}$ 44.3 mg/L, SS 13.2 mg/L, TN 1.4 mg/L, TP 0.15 mg/L and best practicable control technology currently available (BPT) was neutralization, activated sludge treatment and coagulation and sedimentation for the pulp, paper and paper board manufacturing.

Regulation of Activity of the Response Regulator RssB (Response Regulator RssB의 활성 조절)

  • Park, Hee Jeong;Bang, Iel Soo
    • Korean Journal of Microbiology
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    • v.49 no.3
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    • pp.215-220
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    • 2013
  • Against environmental stresses, many bacteria utilize the alternate sigma factor RpoS that induces transcription of the specific set of genes helpful in promoting bacterial survival. Intracellular levels of RpoS are determined mainly by its turnover through proteolysis of ClpXP protease. Delivery of RpoS to ClpXP strictly requires the adaptor protein RssB. The two-component-type response regulator RssB constantly interacts with RpoS, but diverse environmental changes inhibit this interaction through modification of RssB activity, which increases RpoS levels in bacteria. This review discusses and summarizes recent findings on regulatory factors in RssB-RpoS interactions, including IraD, IraM, IraP anti-adaptor proteins of RssB and phosphorylation of N-terminal receiver domain of RssB. New information shows that the coordinated regulation of RssB activity in controlling RpoS turnover confers efficient bacterial defense against stresses.

The Study on the Comparison of the ISCST3 Model and Receptor Model by Dispersion Tracing of Particulate Matter from Large Scale Pollution Sources (대단위배출원에서 기인한 입자상오염물질의 확산ㆍ추적을 통한 ISCST3모델과 수용모델의 비교연구)

  • 전상기;이성철;박경선
    • Journal of Korean Society for Atmospheric Environment
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    • v.19 no.6
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    • pp.789-803
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    • 2003
  • The purpose of this study is to compare the usefulness between Gaussian dispersion model and receptor model with the experimental result of the dispersion tracing of the particulate pollutants from Taean coal-fired power plants. For this purpose, the component analysis of the collected PM 10 samples was performed. In order to trace the pollution sources, factor analysis was done with the result of the component analysis. As a result of the correlativity analysis of the fifteen power plants' profiles offered by US EPA, the correlativity of No.11202 source profile showed highest rate up to 84.5%. Thus it was adopted as proper one and the contribution rate by each pollution source was calculated by Chemical Mass Balance (CMB)-8 model. The contribution rate, which was the effect rate of the power plants on each measuring point, were calculated with a range of 24∼52% and the standard error was below 0.9 $\mu\textrm{g}$/㎥. This indicates the selection of the source profile was appropriate. Also, the concentrations of each point were calculated by the ISCST3 which is suggested by US EPA as one of the regulatory Gaussian dispersion model. The calculation result showed that the predicted concentration was 50∼58 $\mu\textrm{g}$/㎥, comparing with the measured result of 9∼65 $\mu\textrm{g}$/㎥. It was found that the concentration calculated by ISCST3 was underpredicted. It was thought that the receptor model was more favorable than the Gaussian dispersion model in estimating the effect of the particulate matter on a certain receptive point.

Effects of Postnatal Exposure to Octylphenol on the Transcriptions of Steroidogenic Enzymes in Mouse Testis

  • Kim, Suel-Kee;Lee, Ho-Joon;An, Su-Yeon;Lee, Chang Joo;Yoon, Yong-Dal
    • Korean Journal of Environmental Biology
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    • v.22 no.4
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    • pp.550-558
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    • 2004
  • The effects of postnatal exposure to octylphenol(OP) on the expressions of the steroidogenic enzymes and testosterone production were evaluated. Postnatal male mice (15-day-old) were injected with 2 or 20mg $kg^{-l}$ body weight (BW) of OP for 5 days and sacrificed on postnatal day 21. Testosterone concentration was measured by radioimmunoassay and the expressions of the testicular genes were determined by RT-PCR analyses. Significant reductions in the mean body and testis weight were observed in the OP treated animals. No marked alteration in the histological structure of the testis were observed, however, slight reduction in the seminiferous tubule diameter and the number of Leydig cells and several pyknotic cells could be identified in the 20 mg $kg^{-l}$ BW of the OP treated animals. Serum testosterone concentration was dramatically reduced and the mRNA expressions of the steroidogenic acute regulatory protein (StAR), cholesterol side-chain cleavage enzyme (P450scc) and $17\beta$-hydroxylase/Cl7-20 lyase $(P450_{17\alpha})$ were decreased. No significant changes of the gene expressions of the steroidogenic factor-l (SF-I) and estrogen and androgen receptor after the OP treatment showed that the decreased expressions of the steroidogenic enzymes in the present study did not correlate with these genes. Altogether, the present study demonstrates that postnatal treatment of OP inhibits steroidogenesis by decreasing the transcriptional expressions of the StAR and steroidogenic enzymes. The alteration in steroidogenesis may adversely affect the normal development of the testis and sper- matogenesis.

Modulation of $GABA_A$ Receptor by Protein Kinase C in Autonomic Major Pelvic Ganglion Neurons

  • Choi, Yeun-Jong;Cha, Seung-Kyu;Kim, Dae-Ran;Kong, In-Deok
    • Biomedical Science Letters
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    • v.14 no.2
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    • pp.69-76
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    • 2008
  • ${\gamma}$-aminobutyric acid (GABA) is the major inhibitory neurotransmitter in the central nervous system, and its actions are mediated by subtypes of GABA receptors named as $GABA_A$, $GABA_B,\;and\;GABA_C,\;GABA_A$, receptor consisting of ${\alpha},\;{\beta},\;{\gamma}\;and\;{\delta}$ subunits is a heterooligomeric ligand-gated chloride channel. This study was performed to investigate regulation of $GABA_A$ receptor by protein kinase C(PKC). Ion currents were recorded using gramicidine-perforated patch and whole cell patch clamp. mRNA encoding the subunits of PKC expressed in major pelvic ganglion (MPG) neurons was detected by using RT-PCR. The GABA-induced inward current was increased by PKC activators and decreased by PKC inhibitors, respectively. These effects were not associated with intracellular $Ca^{2+}$ and GAG (1-oleoyl-2-acetyl-sn-glycerol), a membrane permeable diacylglycerol (DAG) analogue. These results mean that the subfamily of PKC participating in activation of $GABA_A$ receptor would be an atypical PKC (aPKC). Among theses, ${\xi}$ isoform of aPKC was detected by RT-PCR. Taking together, we suggest that excitable $GABA_A$ receptor in sympathetic MPG neuron seemed to be regulated by aPKC, particular in ${\xi}$ isoform. The regulatory roles of PKC on excitatory $GABA_A$ receptors in sympathetic neurons of MPG may be an important factor to control the functional activity of various pelvic organs such as bowel movement, micturition and erection.

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