• Title/Summary/Keyword: U2OS

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Inhibition of c-FLIP by RNAi Enhances Sensitivity of the Human Osteogenic Sarcoma Cell Line U2OS to TRAILInduced Apoptosis

  • Zhang, Ya-Ping;Kong, Qing-Hong;Huang, Ying;Wang, Guan-Lin;Chang, Kwen-Jen
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.6
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    • pp.2251-2256
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    • 2015
  • To study effects of cellular FLICE (FADD-like IL-$1{\beta}$-converting enzyme)-inhibitory protein (c-FLIP) inhibition by RNA interference (RNAi) on sensitivity of U2OS cells to tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL)-induced apoptosis, plasmid pSUPER-c-FLIP-siRNA was constructed and then transfected into U2OS cells. A stable transfection cell clone U2OS/pSUPER-c-FLIP-siRNA was screened from the c-FLIP-siRNA transfected cells. RT-PCR and Western blotting were applied to measure the expression of c-FLIP at the levels of mRNA and protein. The results indicated that the expression of c-FLIP was significantly suppressed by the c-FLIP-siRNA in the cloned U2OS/pSUPER-c-FLIP-siRNA as compared with the control cells of U2OS/pSUPER. The cloned cell line of U2OS/pSUPER-c-FLIP-siRNA was further examined for TRAILinduced cell death and apoptosis in the presence of a pan-antagonist of inhibitor of apoptosis proteins (IAPs) AT406, with or without 4 hrs pretreatment with rocaglamide, an inhibitor of c-FLIP biosynthesis, for 24 hrs. Cell death effects and apoptosis were measured by the methods of MTT assay with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and flow cytometry, respectively. The results indicated that TRAIL-induced cell death in U2OS/pSUPER-c-FLIP-siRNA was increased compared with control cells U2OS/pSUPER in the presence or absence of AT406. Flow cytometry indicated that TRAIL-induced cell death effects proceeded through cell apoptosis pathway. However, in the presence of rocaglamide, cell death or apoptotic effects of TRAIL were similar and profound in both cell lines, suggesting that the mechanism of action for both c-FLIP-siRNA and rocaglamide was identical. We conclude that the inhibition of c-FLIP by either c-FLIP-siRNA or rocaglamide can enhance the sensitivity of U2OS to TRAIL-induced apopotosis, suggesting that inhibition of c-FLIP is a good target for anti-cancer therapy.

uPC player : An OS Virtualization-based Technique to Support Stateless Computing Execution Environment on Windows (uPC player : 윈도우 운영체제에서의 OS가상화 기반 무상태 컴퓨팅 실행환경 지원 기술)

  • Sung, Baek-Jae;Park, Chan-Ik
    • Journal of KIISE:Computing Practices and Letters
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    • v.16 no.1
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    • pp.125-129
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    • 2010
  • Stateless computing supports a mobility of computing environment easily. It is becoming a major technology for securing personal user's information on shared computing environment. With the advance of virtualization technology and cloud computing, stateless computing is an essential part of personal computing environment connectivity (user's setting and data is stored in remote server or some storage, and it can be restored at any computing environment) In this paper, we propose uPC player that supports stateless computing execution environment on Windows. uPC player provides Windows operating system to user by using an uPC OS virtualization module. In this paper, we leverage how uPC player is designed and implemented for supporting a stateless computing execution environment. uPC player provides a desktop switch between host-system execution environment and uPC virtual execution environment. And it needs just one second for loading uPC virtual execution environment by using OS virtualization-based technique.

Suppression of Ku80 Correlates with Radiosensitivity and Telomere Shortening in the U2OS Telomerase-negative Osteosarcoma Cell Line

  • Hu, Liu;Wu, Qin-Qin;Wang, Wen-Bo;Jiang, Huan-Gang;Yang, Lei;Liu, Yu;Yu, Hai-Jun;Xie, Cong-Hua;Zhou, Yun-Feng;Zhou, Fu-Xiang
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.2
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    • pp.795-799
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    • 2013
  • Ku70/80 heterodimer is a central element in the nonhomologous end joining (NHEJ) DNA repair pathway, Ku80 playing a key role in regulating the multiple functions of Ku proteins. It has been found that the Ku80 protein located at telomeres is a major contributor to radiosensitivity in some telomerase positive human cancer cells. However, in ALT human osteosarcoma cells, the precise function in radiosensitivity and telomere maintenance is still unknown. The aim of this study was to investigate the effects of Ku80 depletion in the U2OS ALT cell line cell line. Suppression of Ku80 expression was performed using a vector-based shRNA and stable Ku80 knockdown in cells was verified by Western blotting. U2OS cells treated with shRNA-Ku80 showed lower radiobiological parameters (D0, Dq and SF2) in clonogenic assays. Furthermore, shRNA-Ku80 vector transfected cells displayed shortening of the telomere length and showed less expression of TRF2 protein. These results demonstrated that down-regulation of Ku80 can sensitize ALT cells U2OS to radiation, and this radiosensitization is related to telomere length shortening.

Transcriptional Activation and Repression of Cell Cycle Regulatory Molecules by Trichostatin A (Trichostatin A 처리에 의하 세포주기 조절인자들의 전사활성화 및 불활성화)

  • Baek Jong-Soo;Lee Hee-Kyung;Cho Young-Su;Kim Sung-Young;Park Kwan-Kyu;Chang Young-Chae
    • Journal of Life Science
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    • v.15 no.6 s.73
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    • pp.994-1004
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    • 2005
  • The dihydrofolate reductase (dhfr) promoter contains cis-acting element for the transcription factors Spl and E2F. Transcription of dhfr gene shows maximal activity during the Gl/S phase of cell cycle. The member of the Spl transcriptional factor family can act as both negative and positive regulators of gene expression. There was a report that Spl-Rb and E2F4-pl30 complexes cooperate to establish stable repression of dhfr gene expression in CHOC400 cells. Here, we examined the role of HDAC in dhfr, cyclin E, and cyclin A gene regulation using the histone deacetylation inhibitor, trichostatin A (TSA) in U2OS and C33A cells, a Rb-positive human osteosarcoma cell line, and a Rb-negative cervical carcinoma cell line, respectively. When the dhfr promoter constructs were applied in U2OS cells, TSA markedly stimulated over 14-fold of dhfr promoter activity through dhfr-Spl sites by the deletion of an E2F element. In contrast, the deletion of dhfr-Spl binding sites completely abolished promoter stimulation by TSA. The dhfr promoter activity including dhfr-Spl sites increased only 2-fold in C33A cells. Promoter activity containing only dhfr-E2F site did not have much effect by the treatment of TSA in both U2OS and C33A cells. On the other hand, treatment with TSA induced significantly mRNA expression of dhfr and cyclin E, whereas levels of cyclin A decreased in U2OS cells, but had no effect in C33A cells. These results indicate that TSA have contradictory effect, activation of dhfr and cyclin E genes on Gl phase, and down-regulation of cyclin A on G2 phase through transcriptional regulation in U2OS cells.

Effect of Ginsenosides from Panax ginseng on Proliferation of Human Osteosarcoma Cell $U_2OS$

  • Deqiang Dou;Jie Ren;Yingjie Chen;Youwei Zhang;Xinsheng Yao
    • Proceedings of the Ginseng society Conference
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    • 2002.10a
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    • pp.376-384
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    • 2002
  • Object To find out which of the 27 ginsenosides isolated from Panax ginseng C.A. Mey that may inhibit the proliferation of human osteosaocoma cell line $U_2OS$. Methods Effects of each individual ginsenoside on the proliferation of $U_2OS$ cell were studied by determining the viability of cancer cells during culture with or without the presence of the test compound. DNA assay was determined by flow cytometry. Results Ginsonosides -Ro, $-Rh_l,\;-Rh_2,\;-F_1\;and\;-L_8$ at concentrations of 5 ,umol/L could obviously suppress the proliferation of $U_2OS$ cells while ginsenosides $-Rg_1,\;-F_3,$ -Rf, PPT and PT significantly inhibited the cancer cells. Flow cytometry revealed that ginsenosides $-Ro,-Rg_1-Rf,-F_1-Rh_2,PPT$ and PT induced cell cycle arrest at $G_0/G_1$ phase with obvious decrease of cell count at Sand $G_2+M$ phase, Moreover, ginsenosides $-Rf_1,-Rg_1,\;-F_1$ and PPT induced significantly high rates of cell death as compared with the control. Conclusion These data suggested that ginsenosides inhibited $U_2OS$ proliferation Via cell cycle arrest or induction of cell death.

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Jab1 has negative effects on p53-mediated genotoxic stresses

  • Lee, Eun-Woo;Lee, Sang-Sik;Song, Jae-Whan
    • BMB Reports
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    • v.42 no.5
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    • pp.299-303
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    • 2009
  • In this study, we use promoter analysis to show that interaction between Jab1 and p53 induces suppression of p53 activation in U2OS and H1299 cells. Interaction between p53 and Jab1 was further confirmed by immunoprecipitation and immunofluorescent analyses. In particular, Jab1 was able to induce nuclear export of p53 as previously reported. When Jab1 was overexpressed in U2OS cells followed by etoposide or hydrogen peroxide ($H_2O_2$), cell death induced by such stresses was protected against. On the contrary, when the level of Jab1 was suppressed in U2OS cells, cytotoxicity imposed by etoposide and $H_2O_2$ was dramatically increased, suggesting a cell protective role for Jab1. These results indicate that Jab1 is a negative regulator of p53 and a plausible oncogene.

A Comparison Study of MMP Inhibitors' and Doxorubicin's Effects on the Apoptosis of U2OS Osteosarcoma Cell Line (U2OS 골육종 세포주의 세포자멸사에서 MMP억제제와 Doxorubicin 작용의 비교연구)

  • Moon, Jeong-Seok;Yeom, Bum-Woo
    • The Journal of the Korean bone and joint tumor society
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    • v.13 no.2
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    • pp.88-95
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    • 2007
  • Purpose: The purpose of this study was to compare the proapoptotic effects of matrix metal-loproteinase inhibitor (MMPI) and doxorubicin on wild-type p53 osteosarcoma cell line, socalled U2OS cell line. Materials and Methods: U2OS cells were treated with MMP inhibitor III (MMPI III) and doxorubicin, either respectively or simultaneously. In cells treated with doxorubicin, Fas-neutralizing antibody so called ZB4 was additionally treated to examine whether the doxorubicin played a role through the Fas/FasL pathway. Cells were analysed regarding to apoptosis and cell death by flow cytometry. Results: U2OS cells incubated with doxorubicin showed significant amount of cell death in dose-dependent manner. However, those incubated with MMPI III mostly remained viable state. In addition, there is no relationship between two drugs. Cells treated with doxorubicin and ZB4 at the same time did not show down regulation of apoptosis through inhibition of Fas/FasL pathway. Conclusion: It is important to re-examine MMP inhibitor's effect on other osteosarcoma cell line with wild-type p14 as well as wild-type p53 to evaluate its proapoptotic effect.

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Study for the standardization of Os Draconis and comparison composition before and after using processed method on Os Draconis (용골(龍骨)(부용치)(附龍齒)의 품질표준(品質標準) 및 포제전후의 성분비교(成分比較))

  • Lee Jang-Cheon
    • Herbal Formula Science
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    • v.11 no.1
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    • pp.171-195
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    • 2003
  • Objectives: This experimental study has been done to compare the Os Draconis composition before and after using processing method. Os Draconis has a quality for calming the liver meridian function and relaxation the mind. Methods: I studied the Os Draconis and processed Os Draconis by vinegar to compare the compositions and its' character. Results: Os Draconis is not a dinosaur's bone fossil but a mammal's bone fossil which has a Calcite mineral, an Apatite mineral, $SiO_2\;Al_2$ O, etc. Os Draconis contains a main ingredients CaO>50.00%. Processed Os Draconis which is heated and soaked in vinegar changes to weak condition Conclusion: Os Draconis is supposed to be a mammal's bone fossil. Some Os Draconis has a radioactive substance like a U, Th so we pay heed to deal with it in a medical clinic.

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A Study on Turbulent Characteristics of Turbulent Pulsating Flows in a Square Duct (4각 덕트내에서 난류 맥동유동의 난류특성에 관한 연구)

  • Park, G.M.;Go, Y.H.
    • Korean Journal of Air-Conditioning and Refrigeration Engineering
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    • v.2 no.3
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    • pp.188-198
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    • 1990
  • Turbulent characteristics of turbulent pulsating flows were studied experimentally in a square duct. Velocity waveforms, velocity profiles, and turbulent intensity of turbulent pulsating flow were investigated by using a hot-wire anemometer with data acquisition and a processing system in a square duct with a ratio of 1 ($40mm{\times}40mm$) to 4,000mm long. Turbulent components were shown to be larger in decelerating than in accelerating regions and also larger for a large phase of velocity and U'rms distribution of turbulent flow. The effect of velocity amplitude ratio does not exist for specified time [${\theta}(z^{\prime})$], amplitude ratio (${\mid}U^{\prime}_{rms.os.1}{\mid}/{\mid}U_{m.os.1}{\mid}$), and phase difference (${\Delta}U^{\prime}_{rms.os.1}-{\Delta}U_{m.os.1}$) in either turbulent oscillating or cross-sectional mean velocity components. The effect of dimensionless angular frequency for specified time [${\theta}(z^{\prime})$] can be disregarded because the dimensionless angular frequency does not affect the specified time. The velocity distributions of turbulent pulsating flows for various time-averaged Reynolds numbers are in approximate agreement with the velocity distributions for equivalent Reynolds numbers and 1/7th power law of steady flow.

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Executable Code Sanitizer to Strengthen Security of uC/OS Operating System for PLC (PLC용 uC/OS 운영체제의 보안성 강화를 위한 실행코드 새니타이저)

  • Choi, Gwang-jun;You, Geun-ha;Cho, Seong-je
    • Journal of the Korea Institute of Information Security & Cryptology
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    • v.29 no.2
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    • pp.365-375
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    • 2019
  • A PLC (Programmable Logic Controller) is a highly-reliable industrial digital computer which supports real-time embedded control applications for safety-critical control systems. Real-time operating systems such as uC/OS have been used for PLCs and must meet real-time constraints. As PLCs have been widely used for industrial control systems and connected to the Internet, they have been becoming a main target of cyberattacks. In this paper, we propose an execution code sanitizer to enhance the security of PLC systems. The proposed sanitizer analyzes PLC programs developed by an IDE before downloading the program to a target PLC, and mitigates security vulnerabilities of the program. Our sanitizer can detect vulnerable function calls and illegal memory accesses in development of PLC programs using a database of vulnerable functions as well as the other database of code patterns related to pointer misuses. Based on these DBs, it detects and removes abnormal use patterns of pointer variables and existence of vulnerable functions shown in the call graph of the target executable code. We have implemented the proposed technique and verified its effectiveness through experiments.