• Title/Summary/Keyword: actinomycin D

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Identification and Functional Analysis of LsMNPV Anti-apoptosis Genes

  • Kim, Yu-Sin;Xiao, Hua-Zhong;Du, En-Qi;Cai, Guo-Shuai;Lu, Song-Ya;Qi, Yi-Peng
    • BMB Reports
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    • v.40 no.4
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    • pp.571-576
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    • 2007
  • Three anti-apoptosis genes, Ls-iap2, iap3 and p49 were found in Leucania separata multiple nuclear polyhedrovirus. Amino acid sequence homology of Ls-IAP2 and Ls-IAP3 with Op-IAP2 and Op-IAP3 from Orgyia pseddotsugata MNPV were 20% and 42%, while that of Ls-P49 is 28% with Sl-P49 from Spodoptera littorolis MNPV. Ls-IAP2 contains one baculoviral IAP repeat (BIR) domain followed by a RING domain, while Ls-IAP3 contains two BIRs and a RING. Ls-P49 contains a reactive site loop, predicted cleavage site (KKLD$^{74}{\downarrow}$G) that is different from Sl-P49 (TVID$^{94}{\downarrow}$G). Expressed Ls-iap3 or Ls-p49 under presence of actinomycin D in SF9 cells, DNA ladder assayrevealed that Ls- IAP3 or Ls-P49 could block the apoptosis of SF9 cells induced by actinomycin D. Replication of p35 deficient-mutant Autographa californica MNPV in SF9 cells was also rescued when Ls-iap3 or Ls-p49 was expressed transiently. No anti-apoptotic activity was observed for Ls-IAP2. The results showed that both of Ls-IAP3 and Ls-P49 were functional apoptotic suppressors in SF9 cells.

THE EFFECTS OF INSULIN-LIKE GROWTH FACTOR I (IGF-I) ON EXPRESSION OF VASCULAR ENDOTHELIAL GROWTH FACTOR (VEGF) MRNA IN MG-63 OSTEOBLASTLIKE CELLS (MG-63 세포주에서 Vascular Endothelial Growth Factor (VEGF) mRNA 발현에 대한 Insulin-like Growth Factor I (IGF-I)의 효과에 대한 연구)

  • Suh, Je-Duck;Myung, Hoon;Kang, Nara;Choung, Pill-Hoon
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.31 no.5
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    • pp.363-369
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    • 2005
  • Purpose: To determine the role of Insulin-like Growth Factor-I (IGF-I) in the regulation of Vascular Endothelial Growth Factor (VEGF) expression in MG-63 cells and then to find the mechanism b which this regulation occurs. Materials and methods: MG-63 cells were grown to confluence in 60-mm dishes. To determine the effects of IGF-I on expression of VEGF mRNA according to time and concentration, the cells were treated with 10 nM IGF-I, following isolation of total RNA and Northern blot analysis after 1, 2, 4, 8, 12, 24 hours and after 2 hours of treatment with 0.5, 2, 10, 25, 50 nM IGF-I respectively, isolation of total RNA and Northern blot analysis were followed. To determine the mechanism of action of IGF-I, inhibitors such as hydroxyurea $(76.1\;{\mu}g/ml)$, actinomycin D $(2.5\;{\mu}g/ml)$, cycloheximide $(10\;{\mu}g/ml)$ were added 1 hour after treatment of 10 nM IGF-I. Results: 1. the expression of VEGF mRNA was increased with treatment of IGF-I. 2. The expression of VEGF mRNA was increased according to time-and concentration dependent manner of IGF-I. 3. The effect of IGF-I was decreased by hydroxyuera, actinomycin D, but not by cycloheximide. Conclusion: IGF-I regulate the expression of VEGF mRNA in the level of DNA synthesis and transcription. These results could suggest that IGF-I plays an important role in angiogenesis in the process of new bone formation and remodeling.

Outcomes of Metastatic Gestational Trophoblastic Neoplasia: Fourteen Year Experience from a Northern Thailand Tertiary Care Center

  • Suprasert, Prapaporn;Siriaree, Sitthicha;Manopunya, Manatsawee
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.3
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    • pp.1357-1362
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    • 2016
  • Metastatic gestational trophoblastic neoplasia (GTN) is an uncommon cancer. The principal treatment consists of chemotherapy with or without surgery or radiotherapy. We here retrospectively reviewed the outcomes of metastatic GTN treated at our institute between January, 1999 and December, 2013. Sixty-three patients met the criteria. The median age was 30.0 years and almost 90% were referral cases. Nearly 40% of the studied patients presented with vaginal bleeding while 22.2% were asymptomatic. The most common antecedent pregnancy was hydatidiform mole (57.1%) followed by term pregnancy (20.6%). The median interval time from antecedent pregnancy to the development of GTN was three months and the median pretreatment B-hCG was 58,274 mIU/ml. Stage III (74.6%) was the most common staging followed by stage IV (20.6%) and stage II (4.8%). The most frequent surgery was hysterectomy (31.7%). Thoracotomy and craniotomy were performed in three and two patients, respectively. The most common first line chemotherapy regimen was methotrexate and folinic acid (36.5%) followed by EMA (etoposide, methotrexate, actinomycin D) (34.9%), EMACO (etoposide, methotrexate, actinomycin D, cyclophosphamide, vincristine) (17.5%) with the remission rate of 66.7%. Nearly one-third of the patients were given a subsequent chemotherapy regimen after failure with the first line therapy and showed a final response rate of 73.0%. However, in stage IV, the response to first line treatment was only 38.5%. In conclusion, the outcomes of metastatic GTN were poor especially with the higher stages.

The Mechanisms of Resistance to TNF in TNF-Sensitive Cancer Cells Transfected with TNF-$\alpha$ Gene Using Retroviral Vector (암세포에서 Retroviral Vector를 이용한 종양괴사인자 유전자 이입후 획득된 종양괴사인자 내성의 기전)

  • Lee, Hyuk-Pyo;Oh, Yeon-Mok;Yoo, Chul-Gyu;Kim, Young-Whan;Shim, Young-Soo;Han, Sung-Koo
    • Tuberculosis and Respiratory Diseases
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    • v.44 no.3
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    • pp.547-558
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    • 1997
  • Background : Tumor necrosis factor(TNF) has been considered as an important candidate for cancer gene therapy based on its potent anti-tumor activity. However, since the efficiency of current techniques of gene transfer is not satisfactory, the majorities of current protocols is aiming the in vitro gene transfer to cancer cells and re-introducing genetically modified cancer cells to hoot. In previous study, it was shown that TNF-sensitive cancer cells transfected with TNF-$\alpha$ cDNA would become highly resistant to TNF. Understanding the mechanisms of TNF-resistance in TNF-$\alpha$ gene transfected cancer cells would be an important step for improving the efficacy of cancer gene therapy as well as for better understandings of tumor biology. This study was designed to evaluate the role of new protective protein synthesis in the acquired resistance to TNF of TNF-$\alpha$ gene transfected cancer cells. Method : We transfected TNF-$\alpha$ c-DNA to WEHI164, a murine fibrosarcoma cell line, using retroviral vector(pLT12SN(TNF)) and confirm the expression of TNF with PCR, ELISA, MIT assay. Then we determined the TNF resistance of TNF gene transfected cells(WEHI164-TNF) and the changes of TNF sensitivities after treatments with actinomycin D(transcription inhibitor) and cycloheximide ( translation inhibitor). Results : WEHI164 which was sensitive to TNF became resistant to TNF after being transfected with TNF-$\alpha$ gene and the resistance to TNF was partially reversed after treatment with actinomycin D, but not with cycloheximide. Conclusion : The acquired resistance to TNF after TNF-$\alpha$ gene transfection may be associated with synthesis of some protective proteins.

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Structure Elucidation of a Potent Anti-MRSA Antibiotic, AM3, Produced by Streptomyces sp. (방선균에 의해 생산된 항 MRSA 항생물질 AM3의 구조 연구)

  • Lim, Yoong-Ho;Chang, Jun-Hwan;Kim, Jong-Hoon;Suh, Jung-Woo;Jung, Jae-Kyung;Lee, Chul-Hoon
    • Applied Biological Chemistry
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    • v.38 no.6
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    • pp.516-521
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    • 1995
  • In order to find a potent anti-methicillin resistant Staphylococcus aureus (MRSA) antibiotic, actinomycetes isolated from the samples collected in Korean marine silt were screened. From the culture broth of the isolated Streptamyces strain AM045, a substance showing excellent biological activity against MRSA was found, isolated and named AM3. The compound showed strong activities against MRSA, S. epidermidis, E. faecium and E. faecalis, which were better than those of vancomycin and teicoplanin. Unfortunately, AM3 was identified as Actinomycin V. However, this paper reports the three dimensional study of AM3 based on high resolution nmr and Computer Aided Molecular Modeling(CAMM), and the fact that the structure of the pentapeptide lactone ring with oxo-proline in chloroform solution does not have 'C conformation' any more.

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Effect of Copper on the Regulation of Ferroportin-1 Gene Expression (구리가 Ferroportin-1 유전자 발현 조절에 미치는 영향)

  • Park, Bo-Yoen;Chung, Ja-Yong
    • Journal of Nutrition and Health
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    • v.42 no.5
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    • pp.434-441
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    • 2009
  • Ferroportin-1 (FPN) is a transporter protein that is known to mediate iron export from macrophages. The purpose of this study was to investigate the effect of copper on the regulation of FPN gene expression in J774 mouse macrophage cells. J774 cells were treated with various concentrations of $CuSO_4$ and RT-PCR analyses were performed to measure the steady-state levels of mRNAs for FPN and divalent metal transporter 1 (DMT1, an iron importer). Copper treatment significantly increased FPN mRNAs in a dose-dependent manner, but didn't change the levels of DMT1 mRNA. Experiments with transcriptional inhibitor actinomycin D (0.5 ${\mu}g$/mL) revealed that copper treatment did not affect the half-life of FPN mRNAs in J774 cells. On the other hand, results from luciferase reporter assays showed that copper directly stimulated the promoter activity of FPN. In summary, our data showed copper induced FPN mRNA of macrophages via a transcriptional rather than post-transcriptional mechanisms.

Effect of Carbon Tetrachloride on the Changes of Xanthine Oxidase Activity in Rate Previously Fed Low or High Protein Diet (식이성 단백질 함량에 따른 흰쥐에 사염화탄소 투여가 Xanthine Oxidase활성에 미치는 영향)

  • 윤종국;이상일;신중규
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.20 no.6
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    • pp.527-537
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    • 1991
  • To evaluate an effect of liver xanthine oxidase on the induction of liver damage, carbon tetrachloride (CCl4) was intraperitoneally injected twice at 0.1ml/100g body weight to the rate fed a low (LP)or high protein diet(HP) while the control group fed LP or HP received only olive oil. The changing rate of liver xanthine oxidas activity was compared with that of a free radical generating enzyme, liver aniline hydroxylase and a scavenging enzyme, glutathions S-transferase activity between the rate fed a LP and those fed HP, and the two groups treated with CCl4. Concomitantly, the degree of liver damage which could be considered as the paramete for CCl4 metabolism in case of CCl4-intoxicated animal was observed in the present experimental conditions and the effect of allopurinol, xanthine oxidase inhibitor, on the CCl4-toxicity of rate liver was alos demostrated. On the other hand, the comparative effect of actinomycin D on the liver and serum xanthine oxidase of CCl4-treated rats fed HP with that of those fed LP and the kinetics of purifed liver enzyme from the liver of CCl4-treated rats fed HP was also compared with that of those fed LP to clarify the differences of xanthine oxidase activity between two groups. The increasing rate of liver weigth/body wt, serum levels of ALT and the decreasing rate of hepatic ALT activity and protein contents to each control group were higher in CCl4-treated rats fed HP than those fed LP. Under the animal models as indentified by the present data herein, the liver xanthine oxidase activity was higher in CCl4-treated rats fed HP than those fed LP, and the control group fed HP also showed the much higher activity xanthine oxidase than that fed LP, whereas there were no differences in the activity of hepatic aniline hydroxylase and glutathions S-transferase between the two group treated with CCl4. Although the hepatic aniline hydroxylase activity was somewhat higher in the rats fed HP than those fed LP, the increasing rate of liver xanthine oxidase to the rats fed LP was higher in those fed HP than that of liver aniline hydroxylase. The degree of liver damage identified such as liver weight and serum ALT activity was less in the CCl4-treated rats pretreated with allopurinol. These results suggest that even a system at which xanthine oxidase acts as well as the drug metabolizing enzyme may influence the acelatin of CCl4 metabolism. In addition, the purified liver xanthine oxidase from CCl4-treated rats fed HP showed decreased Km value when compared to its control group. The Km value of liver xanthine oxidase of CCl4-treated rats fed LP showed a similar Km value with its control group. Furthermore, the decreasing rate of liver and serum xanthine oxidase acitivity in CCl4-treated rats pretreated with actinomycin D to the CCl4-treated rats was higher in rats fed HP than in those fed LP. These results suggest that the inductino of xanthine oxidase in CCl4-treated rats fed HP may be greater than in those fed LP.

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Transcriptional activation of an anti-oxidant mouse Pon2 gene by dexamethasone

  • Lim, Ji-Ae;Kim, Sang-Hoon
    • BMB Reports
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    • v.42 no.7
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    • pp.421-426
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    • 2009
  • Glucocorticoids regulate multiple physiological processes such as metabolic homeostasis and immune response. Mouse Pon2 (mPon2) acts as an antioxidant to reduce cellular oxidative stress in cells. In this present study, we investigated the transcriptional regulation of mPon2 by glucocorticoids. In the presence of glucocorticoid analogue dexamethasone, the expression of mPon2 mRNA in cells was increased, whereas the expression was inhibited by a transcription inhibitor actinomycin D. Glucocorticoid receptors bound to the putative glucocorticoid response elements located between -593 bp and -575 bp of the mPon2 promoter. Transcriptional activity was completely blocked when the putative element was mutated. Taken together, these results suggest that the expression of the mPon2 gene is directly regulated by glucocorticoid-glucocorticoid receptor complexes.

Primary mediastinal endodermal sinus tumor [yolk sac tumor]: report of a case (원발성 종격동 내배엽동종 [난황난종]: 1 치험례 보)

  • 장병철
    • Journal of Chest Surgery
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    • v.17 no.3
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    • pp.497-504
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    • 1984
  • The malignant germ cell tumor found in the gonad can originate in the anterior mediastinum. Endodermal sinus tumor[Yolk sac tumor] is a kind of malignant germ cell tumor and is derived from extra-embryonic mesoderm. We experienced a case of primary mediastinal endodermal sinus tumor occurred in 22 year old male patient. His chief complaint was anterior chest pain for 2 days. The tumor located in the anterior mediastinum and invaded upper lobe of the deft lung and pericardium. A left upper lobe resection including phrenic nerve and pericardium was performed and the tumor in the anterior mediastinum was excised. The patient has been treated with combination chemotherapy[Cis-platinum, Vincristine, Actinomycin-D, & Cyclophosphamide} and followed up for 4 months with partial remission.

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Effect of Carbon Tetrachloride on the Changes of Guanase Activity in-Rats Fed Low or High Proteins Diet (食餌性 蛋白質含量에 따른 흰쥐에 사염화탄소 投與가 Guanase 活性變動에 미치는 영향)

  • Kang, Hoe-Yang
    • Journal of Environmental Health Sciences
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    • v.14 no.1
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    • pp.87-101
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    • 1988
  • The effect of hepatic injury produced by CCL, was studied on rats receiving a low protein-high carbohydrate (7% casein), standard protein (20% casein) and a high protein diet (30% casein). The rats fed low protein diet are resistant to CCl$_4$ in its effects on the liver as judged by histology, serum enzymes(guanase, ALT) and the content of hepatic protein. On the other hand, the pretreatment of hydrocortisone before injection of CCl$_4$ to the rats fed a standard diet, slightly decreased both serum ALT and guanase activities. In the pretreatment of actinomycin D, the liver and serum guanase activities were significantly decreased. It indicates that the cause of increasing serum guanase is based on the alteration of membrane permeability and the result of accelerated enzyme synthesis in liver cells of CCl$_4$ intoxicated rats.

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