• 제목/요약/키워드: Cellular transformation

검색결과 133건 처리시간 0.03초

Production and Prophylactic Efficacy Study of Human Papillomavirus-like Particle Expressing HPV16 L1 Capsid Protein

  • Park, Jie-Yun;Pyo, Hyun-Mi;Yoon, Sun-Woo;Baek, Sun-Young;Park, Sue-nie;Kim, Chul-Joong;Haryoung Poo
    • Journal of Microbiology
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    • 제40권4호
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    • pp.313-318
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    • 2002
  • To perform the prophylactic study of a vaccine derived from human papillomavirus (HPV) using Balb/c mice, we produced virus like particles consisting of HPV capsid protein L1 which has been reported to induce significant humoral and cellular immunity using various animal model systems. In order to produce HPV16 VLPs, the cDNA of L1 capsid protein in HPV type 16, obtained by polymerase chain reaction, was inserted into yeast expression vector, YEG$\alpha$-HIR525 under the control of GAL10 promoter. The transformation of YEG$\alpha$-HPV16 L1 was performed into the yeast Saccharomyces cerevisiae Y2805 by the lithium acetate method and the yeast clone expressing the highest level of L1 capsid protein of human papillomavirus type 16 was selected by Western blot analysis using anti-HPV16 L1 antibody. The purification of HPV16 VLP has been performed by the ultracentrifugation and gel-filtration methods. To validate the vaccine efficacy of the purified HPV16 VLPs and investigate the properties of HPV16 VLPs to induce humoral immunity, ELISA assay was performed. A significantly increased production of anti-HPV16 VLP antibodies was observed in sera from immunized mice. The neutralization activity of antibodies in the sera from the vaccinated mice was demonstrated by a rapid and simple assay to detect hemagglutihation inhibition activity.

Relationship between Sloan-Kettering Virus Expression and Granulosa Cells of Atretic Follicles in the Rat Ovary

  • Kim, Hyun;Matsuwaki, Takashi;Yamanouchi, Keitaro;Nishihara, Masugi;Kim, Sung-Woo;Ko, Yeoung-Gyu;Yang, Boh-Suk
    • Reproductive and Developmental Biology
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    • 제35권3호
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    • pp.341-348
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    • 2011
  • Sloan-Kettering virus gene product of a cellular protooncogene c-Ski is an unique nuclear pro-oncoprotein and belongs to the Ski/Sno proto-oncogene family. Ski plays multiple roles in a variety of cell types, it can induce both oncogenic transformation and terminal muscle differentiation when expressed at high levels. The aim of the present study was to locate Ski protein in rat ovaries in order to predict the possible involvement of Ski in follicular development and atresia. First, expression of c-Ski mRNA in the ovaries of adult female rats was confirmed by RT-PCR. Then, ovaries obtained on the day of estrus were subjected to immunohistochemical analysis for Ski and proliferating cell nuclear antigen (PCNA) in combination with terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL). Ski was expressed in granulosa cells that were positive for TUNEL, but negative for PCNA, regardless of the shape and size of follicles. Expression of Ski in TUNEL-positive granulosa cells, but not in PCNA-positive granulosa cells, was also verified in immature hypophysectomized rats having a single generation of developing and atretic follicles by treatment with equine chorionic gonadotropin (eCG). These results indicate that Ski is profoundly expressed in the granulosa cells of atretic follicles, but not in growing follicles, and suggest that Ski plays a role in apoptosis of granulosa cells during follicular atresia.

A Role of Natural Killer Cell in Mouse Infected Herpes Simplex Virus (Herpes Simplex Virus에 감염된 Mouse의 NK세포역할)

  • Lee, Yun-Tai;Lee, Chong-Hoon
    • The Journal of the Korean Society for Microbiology
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    • 제17권1호
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    • pp.7-14
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    • 1982
  • A model of induction of neoplasia by viruses has develpoed from experimental studies in animals and in cultured cells and oncogenic transformation of cells is the result of integration of viral genetic information into the cellular DNA. The evidence for these associations was derived primarily from seroepidemiologic investigation. However, data indicating that the relation between HSV-2 and cervical cancer fits the model derived from experimental animal studies are not yet sufficient to draw conclusion with regard to the etiologic role the virus in the development of the neoplasms. In other hand, the K562 tumor cell is highly susceptible target for natural killer cell lysis by the lymphocytes of human and murine periperal blood. The characteristics of this effector cell type has been investigated. A study on natural killer cell mediated cytotoxicity(NKMC) against $^{51}Cr$-K562 as target cell was studed in HSV-2 infected ICR mouse. We have studied for susceptibility of HSV-2 against mouse embryo fibroblast(MEF) cells and NKMC from HSV-2 infected mouse. The results obtained that the mouse embryo fibroblast cells culture, the number and size of the cells were markedly increased and formed a monolayers relatively rapid, and become complete monolayer sheet around 72 hrs. Duration of cytopathic effect on MEF cells was rapid by serial passing of HSV-2. The morphology of the HSV-2 infected cells appear to be mainly round, ovium, spindle form and some of them was forming large giant cells. The NKMC was decrease in mouse with HSV-2 and comparison between effector/target cells ratio as 25:1 and 50:1 respectively, the NKMC was found to be more significantly decreased than normal control we have concluded that the natural killer cell activity of the viral infected mouse was shown as a suppressed during the HSV-2 infection, day 7th and 14th.

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Color Image Encryption using MLCA and Transformation of Coordinates (MLCA와 좌표변환을 이용한 컬러 영상의 암호화)

  • Yun, Jae-Sik;Nam, Tae-Hee;Cho, Sung-Jin;Kim, Seok-Tae
    • Journal of the Korea Institute of Information and Communication Engineering
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    • 제14권6호
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    • pp.1469-1475
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    • 2010
  • This paper presents a problem of existing encryption methods using pseudo-random numbers based on MLCA or complemented MLCA and proposes a method to resolve this problem. The existing encryption methods have a problem which the edge of original image appear on encrypted image because the image have color similarity of adjacent pixels. In this proposed method, we transform the value and spatial coordinates of all pixels by using pseudo-random numbers based on MLCA. This method can resolve the problem of existing methods and improve the level of encryption by encrypting pixel coordinates and pixel values of original image. The effectiveness of the proposed method is proved by conducting histogram and key space analysis.

STP-C, an Oncoprotein of Herpesvirus saimiri Augments the Activation of NF-κB through Ubiquitination of TRAF6

  • Chung, Young-Hwa;Jhun, Byung-Hak;Ryu, Su-Chak;Kim, Heui-Soo;Kim, Cheol-Min;Kim, Bong-Seok;Kim, Young-Ok;Lee, Sang-Jun
    • BMB Reports
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    • 제40권3호
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    • pp.341-348
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    • 2007
  • Herpesvirus saimiri (HVS), a member of the $\delta$-herpesvirus family, encodes an oncoprotein called Saimiri Transforming Protein (STP) which is required for lymphoma induction in non-human primates. Previous study has shown that STP-C, an oncoprotein of HVS, activates NF-$\kappa$B signaling pathway. However, the detailed mechanism of STP-Cmediated NF-$\kappa$B activation has not been reported yet. We first report that STP-C interacts with TRAF6 protein in vivo and in vitro and further investigation shows that $Glu_{12}$ residue of STP-C is critical for binding to TRAF6. Introduction of ubiquitin together with STP-C augments NF-$\kappa$B activity compared to that of STP-C expression alone. STP-C expression further induces ubiquitination of endogenous TRAF6. In addition, either a deubiquitination enzyme, CYLD or a dominant negative E2-conjugation enzyme reduced NF-$\kappa$B activity in spite of the presence of STP-C, supporting that the interaction between STP-C and TRAF6 induces ubiquitination of TRAF6. NF-$\kappa$B activation by STP-C through the ubiquitinated TRAF6 causes the increased production of IL-8, an inflammatory chemokine and the enhanced expression of costimulatory molecule ICAM, which might ultimately contribute cellular transformation by the exposure of HVS-infected cells with inflammatory microenvironment and chronic activation.

Molecular and Genomic Approaches on Nickel Toxicity and Carcinogenicity

  • Seo, Young-Rok;Kim, Byung-Joo;Ryu, Jae-Chun
    • Molecular & Cellular Toxicology
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    • 제1권2호
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    • pp.73-77
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    • 2005
  • Nickel is the one of potent environmental, the occupational pollutants and the classified human carcinogens. It is a serious hazard to human health, when the metal exposure. To prevent human diseases from the heavy metals, it is seemingly important that understanding of how nickel exerts their toxicity and carcinogenic effect at a molecular and a genomic level. The process of nickel absorption has been demonstrated as phagocytosis, iron channel and diffusion. Uptaked nickel has been suggested to induce carcinogenesis via two pathways, a direct DNA damaging pathway and an indirect DNA damaging pathway. The former was originated from the ability of metal to generate Reactive Oxygen Species (ROS) and the reactive intermediates to interact with DNA directly. Ni-generated ROS or Nickel itself, interacts with DNAs and histones to cause DNA damage and chromosomal abnormality. The latter was originated from an indirect DNA damage via inhibition of DNA repair, or condensation and methylation of DNA. Cells have ability to protect from the genotoxic stresses by changing gene expression. Microarray analysis of the cells treated with nickel or nickel compounds, show the specific altered gene expression profile. For example, HIF-I (Hypoxia-Inducible Factor I) and p53 were well known as transcription factors, which are upregulated in response to stress and activated by both soluble and insoluble nickel compounds. The induction of these important transcription factors exert potent selective pressure and leading to cell transformation. Genes of metallothionein and family of heat shock proteins which have been known to play role in protection and damage control, were also induced by nickel treatment. These gene expressions may give us a clue to understand of the carcinogenesis mechanism of nickel. Further discussions on molecular and genomic, are need in order to understand the specific mechanism of nickel toxicity and carcinogenicity.

Distinctive Features of Advancing Breast Cancer Cells and Interactions with Surrounding Stroma Observed Under the Scanning Electron Microscope

  • Jaafar, Hasnan;Sharif, Sharifah Emilia Tuan;Murtey, Mogana Das
    • Asian Pacific Journal of Cancer Prevention
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    • 제13권4호
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    • pp.1305-1310
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    • 2012
  • Breast cancer cells undergo transformation when they spread into surrounding tissues. Studies have shown that cancer cells undergo surface alterations and interact with the surrounding microenvironment during the invasion process. The aim of the present study was to analyse these cancer cell surface alterations and interactions of cancer cells and stroma. Twenty 1-methyl-1-nitrosourea-induced breast cancer samples taken from five rats were fixed in McDowell-Trump fixative and then washed in 0.1 M phosphate buffer. The samples were then treated with osmium tetroxide before being washed in distilled water and subsequently dehydrated through graded ethanols. The dehydrated samples were immersed in hexamethyldisilazane (HMDS), then following removal of excess HMDS, the samples were air dried at room temperature in a dessicator. The dried samples were mounted onto specimen stubs and coated with gold coater before being viewed under a scanning electron microscope. We detected the presence of membrane ruffles on the surface of cancer cells and the formation of unique surface membrane protrusions to enhance movement and adhesion to the surrounding stroma during the process of invasion. Advancing cancer cells demonstrated formation of lamellipodia and invadopodia. The stroma at the advancing edge was desmoplastic with many collagen fibres laid down near the cancer cells. Our data suggest that all of these abnormalities could act as hallmarks of invasiveness for breast cancer.

Role of Nitric Oxide in Pepsinogen Secretion from Rat Gastric Chief Cells

  • Sung, Dae-Suk;Seo, Dong-Wan;Choi, Don-Woong;Ahn, Seong-Hoon;Hong, Sung-Youl;Lee, Hoi-Young;Han, Jeung-Whan;Lee, Hyang-Woo
    • Biomolecules & Therapeutics
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    • 제7권2호
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    • pp.105-111
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    • 1999
  • Nitric oxide (NO), a cellular messenger synthesized from L-arginine by NO synthase (NOS, EC.1.14.13.39), is considered to be a regulator of gastric secretion. In the present study, the role of NO in the regulation of exocrine secretion was investigated in rat gastric chief cells. Treatment of chief cells with carba-chol resulted in an increase in the arginine conversion to citrulline, the amount of $NO_{x}$, the release of pepsine-gen, and the level of cGMP Especially, carbachol-stimulated increase of arginine to citrulline transformation, the amount of $NO_{x}$, cGMP level and the release of pepsinogen were partially reduced by the natural NOS inhibitor, $N^{G}$-monomethyl-L-arginine (MMA) and $N^{G}$, $N^{G}$-dimethyl-L-arginine (DMA). Furthermore, MMA- and DMA-induced decrease of pepsinogen secretion showed dose-dependent patters. Activation of NOS is one of the early events in receptor-mediated cascade of reactions in gastric chief cells and NO, not completely, but partially mediates gastric secretion. Agonist-stimulated pepsinogen secretion in chief cells has been considered to be mediated in adenosine 3',5'-cyclic monophosphate pathway and/or guanosine 3', 5'-cyclic monophosphate (cGMP) pathway. Taken together, the above results suggest that partial decrease of exocrine secretion following treatment of NOS inhibitor may result from the inactivation of NOS and subsequent guano- late cyclase, and NO/cGMP pathway may play a pivotal role in exocrine secretion.

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A Study on the Reduction of LSP(Line Spectrum Pair) Transformation Time in Speech Coder for CDMA Digital Cellular System (이동통신용 음성부호화기에서의 LSP 계산시간 감소에 관한 연구)

  • Min, So-Yeon
    • Journal of the Korea Academia-Industrial cooperation Society
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    • 제8권3호
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    • pp.563-568
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    • 2007
  • We propose the computation reduction method of real root method that is used in the EVRC(Enhanced Variable Rate Codec) system. The real root method is that if polynomial equations have the real roots, we are able to find those and transform them into LSP. However, this method takes much time to compute, because the root searching is processed sequentially in frequency region. But, the important characteristic of LSP is that most of coefficients are occurred in specific frequency region. So, to reduce the computation time of real root, we used the met scale that is linear below 1kHz and logarithmic above. In order to compare real root method with proposed method, we measured the following two. First, we compared the position of transformed LSP(Line Spectrum Pairs) parameters in the proposed method with these of real root method. Second, we measured how long computation time is reduced. The experimental result is that the searching time was reduced by about 48% in average without the change of LSP parameters.

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Cloning of Bacillus amyloliquefaciens amylase gene using YRp7 as a vector II. Expression of cloned amylase gene in Saccharomyces cerevisiae (YRp7 vector를 이용한 Bacillus amyloliquefaciens amylase gene의 cloning I I. Saccharomyces cerevisiae에서 발현)

  • 서정훈;김영호;전도연;배영석;홍순덕;이종태
    • Microbiology and Biotechnology Letters
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    • 제14권3호
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    • pp.213-218
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    • 1986
  • Hybrid plasmid pEA24, shuttle vector YRp7 carrying amylase gene of Bacillus amyloliquefaciens, was transformed to yeast Saccharomyces cerevisiae, and the expression of B. amyloliquefaciens amylase gene in yeast was investigated. The frequency of transformation to S. cerevisiae DBY747 with YRp7 was increased by treatment of 40% polyethylene glycol (MW 4, 000), PH 7.0, at 3$0^{\circ}C$, and by regeneration used 2% top agar. The amount of cellular amylase activity produced by S. cerevisiae containing pEA24 was 2% of that secreted from B. amyloliquefaciens, but in case of S. cerevisiae transformant, the amylase secreted was not detected. A comparison of genetic stability of pEA24 and YRp7 plasmids in yeast was carried out by cultivation of transformants in tryptophan-supplement-medium. The pEA24 plasmid was more unstable than YRp7 in S. cerevisiae. The size of pEA24 extracted from S. cerevisiae transformants was found to be identical with that from E. coli transformants by agarose gel electrophoresis.

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