• Title, Summary, Keyword: TSA

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Effect of trichostatin A on NF-κB DNA binding activity and myogenesis in C2Cl2 skeletal muscle Precursor cell (C2C12 근육아세포에서 trichostatin A에 의한 NF-κB DNA 결합 활성과 근육발생에 미치는 영향)

  • 임운기;김경창;신혜자
    • Journal of Life Science
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    • v.12 no.1
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    • pp.55-60
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    • 2002
  • The differentiation of skeletal muscle precursor cells in culture is marked by the transcriptional activation of muscle-specific genes and the morphological differentiation of myoblast into multinucleate myotube. In this study, we examined the effect of TSA (Trichostatin A) on WF-kB DNA binding activity and muscle cell fusion in the process of myogenesis. Under TSA treatment, C2C12 myoblast could not fuse to myotube and its NF-kB DNA binding activity was also blocked. To investigate whether these phenomenons were affected by TSA in direct or not, differentiation media (DM) used to differentiate cells without TSA was concentrated and added to C2C12 myoblast with TSA simultaneously. C2C12 myoblast was fused to myotube and NF-kB DNA binding activity was recovered. These results suggest that TSA affects on the differentiation of myoblast, perhaps through several factors, by inhibiting myoblst fusion and blocking NF-kB DNA binding activity.

Preparation and Characteristics of Polyethersulfone Microfiltration Membranes (폴리에테르술폰 정밀여과막의 제조 및 특성 연구)

  • Kim, No-Won
    • Membrane Journal
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    • v.17 no.4
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    • pp.329-337
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    • 2007
  • This is the research about a new method to make the internal separation layer with smallest pore size in polyethersulfone (PES) membrane by adding p-toluenesulfonic acid (TSA) and polyvinylpyrolidone (PVP) to polymeric PES solution. The preparation and morphological characterization of PES sheet membranes containing PVP as a hydrophilic swelling material and TSA as a demixing material were performed. As a result by microflow porometery, the PVP and TSA added PES membranes showed good permeabilities and narrow pore size distributions, comparable to those of the commercial membranes. The concentration of PVP affected the PES characteristics on air permeability and surface structure. The concentration of TSA influenced on pore size distribution but do not affect air permeability. The surface images of FE-SEM shows similar pore size when TSA added or not. However, the cross-section images of FE-SEM show that the TSA added PES membranes have a increase of internal layer thickness with smallest pore size.

A New Member of Human TSA/AhpC as Thioredoxin-dependent Thiol Peroxidase

  • Jeong, Woo-Jin;Cha, Mee-Kyung;Kim, Il-Han
    • BMB Reports
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    • v.33 no.3
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    • pp.234-241
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    • 2000
  • A new type of the human TSA homologous gene was cloned from a HeLa cell cDNA and characterized. The gene product consists of 161 amino acids with a molecular mass of 16,900. The TSA homologous protein, as a new 6th member of the human TSA (hTSA VI), exerted a thioldependent peroxidase activity with the use of thioredoxin system as a physiological electron donor. The values of $V_{max}/K_m$ of hTSA VI for $H_2O_2$ and t-butyl hydroperoxide (t-BOOH) were calculated as $5.53{\times}10^{-2}$ and $3.70{\times}10^{-2}$, respectively. This implies that hTSA VI is a peroxidase, which reduces $H_2O_2$ and t-BOOH. The mutation of $Cys^{47}$ to serine resulted in a complete loss of the peroxidase activity. This suggests that $Cys^{47}$ acts as a primary site of catalysis. The analysis of the tryptic digest derived from hTSA VI revealed that the $Cys^{47}$ exists as a free thiol form. Taken together, these results suggest that the TSA homologous protein is a new type of the human family, which exerts thioredoxin-linked peroxidase activity toward $H_2O_2$ and alkyl hydroperoxide.

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Purification and Characterization of TSA from Lumbricus terrestris (지렁이(Lumbricus terrestris)로부터 Thiol-Specific Antioxidant protein(TSA)의 분리 및 정제에 관한 연구)

  • Kwak, Byung-Koo;Kim, Il-Han;Cha, Mee-Kyung
    • The Journal of Natural Sciences
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    • v.14 no.2
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    • pp.55-65
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    • 2004
  • A thiol-specific antioxidant(TSA) protein was purified from Earthworm, Lumbricus terrestris by DEAE-Cellulose, Phenl sepharose, Sephacryl S-200 gel filtration and HPLC S-300 Column Chromatography. This protein showed a thiol-specific antioxidant activity against inactivation of glutamine synthetase by a metal-catalysed oxidation system capable of generation reaction oxygen species. The molecular mass of the protein was determinated to be 51-kDa by SDS-polyacrylamide gel electrophores. Taken together, the purified TSA protein could be a new member of TSA family.

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Effects of Trichostatin A on In Vitro Development of Porcine Parthenogenetic and Nuclear Transfer Embryos

  • Diao, Yun-Fei;Kenji, Naruse;Han, Rong-Xun;Lin, Tao;Oqani, Reza-K.;Kang, Jung-Won;Jin, Dong-Il
    • Reproductive and Developmental Biology
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    • v.37 no.2
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    • pp.57-64
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    • 2013
  • Developmental potential of cloned embryos is related closely to epigenetic modification of somatic cell genome. The present study was to investigate the effects of applying histone deacetylation inhibitor, trichostatin A (TSA) to activated porcine embryos on subsequent development of porcine parthenogenetic and nuclear transfer embryos. Electrically activated oocytes were treated with 5 nM TSA for different exposure times (0, 1, 2 and 4 hr) and then the activated embryos were cultured for 7 days. The reconstructed embryos were treated with different concentrations of 0, 5, 10 and 25 nM TSA for 1 hr. Also 5 nM TSA was tested with different exposure times of 0, 0.5, 1, 2 and 4 hr. And fetal fibroblast cells were treated with 50 nM TSA for 1, 2 or 4 hr and with 5 nM TSA for 1 hr. Cumulus-free oocytes were enucleated and reconstructed by TSA-treated donor cells and electrically fused and cultured for 6 days. In parthenogenetic activation experiments, 5 nM TSA treatment for 1 hr significantly improved the percentage of blastocyst developmental rates than the other groups. Total cell number of blastocysts in 1 hr group was significantly higher than other groups or control. Similarly, blastocyst developmental rates of porcine NT embryos following 5 nM TSA treatment for 1 hr were highest. And the reconstructed embryos from donor cells treated by 50 nM TSA for 1 hr improved the percentage of blastocyst developmental rates than the control group. In conclusion, TSA treatment could improve the subsequent blastocyst development of porcine parthenogenetic and nuclear transfer embryos.

Diagnosis Value of Membrane Glycolipids Biochemistry Index in Intracranial and Gastrointestinal Tumors

  • Lv, Jun;Lv, Can-Qun;Mei, Ping;Qi, Shi-Mei
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.7
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    • pp.2693-2696
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    • 2015
  • The diagnostic value of membrane glycolipid biochemistry index, the lipid-bound sialic acid (LSA) and total sialic acid (TSA) in cerebrospinal fluid (CSF) was evaluated in 30 intracranial and 65 gastrointestinal tumors. The plasma LSA, TSA and red cell membrane sialic acid (R-SA) in were determined according to the method of Sevenmerhulm. Our results showed that the levels of LSA and TSA in CSF of intracranial tumor patients was higher than that of normal group(p<0.01). The concentration of TSA and LSA in patients with malignant glioma was higher than that of benign meningioma patients(P<0.01). No significance was found between intracranial halmatoma patients and normal control group for levels of membrane glycolipids (p>0.05). Results also found that the plasma LSA, TSA and R-SA of gastric carcinoma were significantly higher than those of control group (p<0.05); while no significant difference was found in the plasma LSA, TSA and R-SA levels between chronic gastritis, gastrohelcoma and normal control group (p>0.05). Plasma LSA, TSA and R-SA levels of gastric carcinoma patient were significantly higher than those of chronic gastritis patients and gastrohelcoma patients(p<0.05). It was also found that plasma LSA, TSA and R-SA contents were significantly higher in large intestine carcinoma patients than in benign in stestine tumor patients (p<0.05) while no significant difference was found between intestine benign tumor and normal control group (p>0.05). The levels of LSA, TSA and R-SA were obviously higher in the patients with metastasis than in the ones without (p<0.05.) The membrane glycolipid biochemistry index LSA and TSA in CSF are sensive markers for diagnosing intracranial tumors. For gastrointestinal malignant tumors the plasma LSA TSA and red blood cell membrane SA may be considered as auxiliary indicators for diagnosis. They can be used for distinguishing benign from malignant tumors.

Antitumor Activity of Histone Deacetylase Inhibitor Trichostatin A in Osteosarcoma Cells

  • Cheng, Dong-Dong;Yang, Qing-Cheng;Zhang, Zhi-Chang;Yang, Cui-Xia;Liu, Yi-Wen
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.4
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    • pp.1395-1399
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    • 2012
  • Background: Histone deacetylase (HDAC) inhibitors have been reported to induce cell growth arrest, apoptosis and differentiation of tumor cells. The present study aimed to examine the effects of trichostatin A (TSA), one such inhibitor, on the cell cycle, apoptosis and invasiveness of osteosarcoma cells. Methods: MG-63 cells were treated with TSA at various concentrations. Then, cell growth and apoptosis were determined by 3-(4, 5-dimethyl-2-thiazolyl)-2H-tetrazolium bromide (MTT) and TUNEL assays, respectively; cell cycling was assessed by flow cytometry; invasion assays were performed with the transwell Boyden Chamber system. Results: MTT assays revealed that TSA significantly inhibited the growth of MG-63 cells in a concentration and time dependent manner. TSA treated cells demonstrated morphological changes indicative of apoptosis and TUNEL assays revealed increased apoptosis of MG-63 cells after TSA treatment. Flow cytometry showed that TSA arrested the cell cycle in G1/G2 phase and annexin V positive apoptotic cells increased markedly. In addition, the invasiveness of MG-63 cells was inhibited by TSA in a concentration dependent manner. Conclusion: Our findings demonstrate that TSA inhibits the proliferation, induces apoptosis and inhibits invasiveness of osteosarcoma cells in vitro. HDAC inhibitors may thus have promise to become new therapeutic agents against osteosarcoma.

Modulacon of Cell Cycle Control by Histone Deacetylase Inhibitor Trichostatin A in A549 Human Non-small Cell Lung Cancer Cells (인체폐암세포 A549의 세포주기 조절인자에 미치는 histone deacetylase inhibitor trichostatin A의 영향)

  • Hwang Ji Won;Kim Young Min;Hong Su Hyun;Choi Byung Tae;Lee Won Ho;Choi Yung Hyun
    • Journal of Life Science
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    • v.15 no.5
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    • pp.726-733
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    • 2005
  • Histone deacetylase (HDAC) inhibitors target key steps of tumor development. They inhibit proliferation, induce differentiation and/or apoptotic cell death, and exhibit potent antimetastatic and antiangiogenic properties in cancer cells in vitro and in vivo. Although they are emerging as a promising new treatment strategy in malignancy, how they exert their effect on human non-small cell lung cancer cells is as yet unclear. The present study was undertaken to investiate the underlying mechanism of a HDAC inhibitor trichostatin A (TSA)-induced growth arrest and its effect on the cell cycle control gene products in a human lung carcinoma cell line A549. TSA treaoent induced the growth inhibition and morphological changes in a concentration-dependent manner. Treatment of A549 cells with TSA resulted in a concentration-dependent increased G1 (under 100 ng/ml) and/or G2/M (200 ng/ml) cell population of the cell cycle as determined by flow cytometry Moreover, 200 ng/ml TSA treatment significantly induced the population of sub-G1 cells (23.0 fold of control). This anti-proliferative effect of TSA was accompanied by a marked inhibition of cyclins, positive regulators of cell cycle progression, and cyclin-dependent kinases (Cdks) expression and concomitant induction of tumor suppressor p53 and Cdk inhibitors such as p21 and p27 Although further studies are needed, these findings provide important insights into the possible molecular mechanisms of the anti-cancer activity of TSA in human lung carcinoma cells.

Inhibition of Histone Deacetylase Activity Diminishes Pressure Overloaded Cardiac Hypertrophy in Mice

  • Hong, Yun-Kyung;Song, Jong-Wook;Lee, Sang-Kil;Lee, Young-Jeon;Rho, Gyu-Jin;Kim, Joo-Heon;Hong, Yong-Geun
    • Reproductive and Developmental Biology
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    • v.35 no.2
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    • pp.159-165
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    • 2011
  • To explore the role of histone deactylase (HDAC) activation in an in vivo model of hypertrophy, we studied the effects of Trichostatin A (TSA). TSA subjected to thoracic aortic banding (TAB)-induced pressure stress in mice. In histological observations, TAB in treated mice showed a significant hypertrophic response, whereas the sham operation remained nearly normal structure with partially blunted hypertrophy. TSA treatment had no effect (measured as HW/BW) on sham-operated animals. TAB animals treated with vehicle manifested a robust ~50% hypertrophic response (p<0.05 vs sham). TAB mice treated with 2 mg/kg/day TSA manifested a blunted growth responses, which was significantly diminished (p<0.05) compared with vehicle-treated TAB mice. TAB mice treated with a lower dose of TSA (0.5 mg/kg/day) manifested a similar blunting of hypertrophic growth (~25% increase in heart mass). Furthermore, to determine activity duration of TSA in vitro, 1 nM TSA was added to H9c2 cells. Histone acetylation was initiated at 4 hr after treatment, and it was peak up to 18 hr, then followed by significantly reduced to 30 hr. We also analyzed the expression of p53 following TSA treatment, wherein p53 expression was elevated at 4 hr, and it was maintained to 24 hr after treatment. ERK was activated at 8 hr, and maintained till 30 hr after treatment suggesting an intracellular signaling interaction between TSA and p53 expression Taken together, it is suggested that HDAC activation is required for pressure-overload growth of the heart. Eventually, these data suggest that histone acetylation may be a novel target for therapeutic intervention in pressure-overloaded cardiac hypertrophy.

Effects of Trichostatin A on Cumulus Expansion during Mouse Oocyte Maturation

  • Du, Ming;Fu, Xiangwei;Zhou, Yanhua;Zhu, Shien
    • Asian-Australasian Journal of Animal Sciences
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    • v.26 no.11
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    • pp.1545-1552
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    • 2013
  • This study was conducted to investigate the effects of Trichostatin A (TSA) on cumulus expansion during mouse oocyte maturation. TSA treatment inhibited cumulus expansion and significantly reduced the cumulus expansion index (CEI) (p<0.05). To determine the underlying mechanism, the expression levels of several key factors that play crucial roles in cumulus expansion including components of extracellular matrix (ECM) (Has2, Ptgs2, Ptx3, and Tnfaip6) and Growth differentiation factor 9 (GDF9) were measured in control and TSA treated samples by real-time PCR. The effect of TSA on ERK phosphorylation (p-ERK1/2) in cumulus cells and GDF9 protein level in fully grown oocytes (FGOs) were detected by Western blotting. The expression levels of the ECM genes were significantly decreased (p<0.05) by TSA treatment while GDF9 expression did not response to TSA (p>0.05). TSA treatment blocked the activation of ERK1/2 (p<0.05) and had no significant effect on GDF9 protein expression (p>0.05). Collectively, these results suggested that TSA treatment altered ECM gene expression and blocked ERK1/2 activation to inhibit cumulus expansion in the mouse.