• 제목/요약/키워드: Sf-21

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SF3B4 Depletion Retards the Growth of A549 Non-Small Cell Lung Cancer Cells via UBE4B-Mediated Regulation of p53/p21 and p27 Expression

  • Kim, Hyungmin;Lee, Jeehan;Jung, Soon-Young;Yun, Hye Hyeon;Ko, Jeong-Heon;Lee, Jeong-Hwa
    • Molecules and Cells
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    • 제45권10호
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    • pp.718-728
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    • 2022
  • Splicing factor B subunit 4 (SF3B4), a component of the U2-pre-mRNA spliceosomal complex, contributes to tumorigenesis in several types of tumors. However, the oncogenic potential of SF3B4 in lung cancer has not yet been determined. The in vivo expression profiles of SF3B4 in non-small cell lung cancer (NSCLC) from publicly available data revealed a significant increase in SF3B4 expression in tumor tissues compared to that in normal tissues. The impact of SF3B4 deletion on the growth of NSCLC cells was determined using a siRNA strategy in A549 lung adenocarcinoma cells. SF3B4 silencing resulted in marked retardation of the A549 cell proliferation, accompanied by the accumulation of cells at the G0/G1 phase and increased expression of p27, p21, and p53. Double knockdown of SF3B4 and p53 resulted in the restoration of p21 expression and partial recovery of cell proliferation, indicating that the p53/p21 axis is involved, at least in part, in the SF3B4-mediated regulation of A549 cell proliferation. We also provided ubiquitination factor E4B (UBE4B) is essential for p53 accumulation after SF3B4 depletion based on followings. First, co-immunoprecipitation showed that SF3B4 interacts with UBE4B. Furthermore, UBE4B levels were decreased by SF3B4 depletion. UBE4B depletion, in turn, reproduced the outcome of SF3B4 depletion, including reduction of polyubiquitinated p53 levels, subsequent induction of p53/p21 and p27, and proliferation retardation. Collectively, our findings indicate the important role of SF3B4 in the regulation of A549 cell proliferation through the UBE4B/p53/p21 axis and p27, implicating the therapeutic strategies for NSCLC targeting SF3B4 and UBE4B.

Effects of a Pre-Filter and Electrolysis Systems on the Reuse of Brine in the Chinese Cabbage Salting Process

  • Kim, Dong-Ho;Yoo, Jae Yeol;Jang, Keum-Il
    • Preventive Nutrition and Food Science
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    • 제21권2호
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    • pp.147-154
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    • 2016
  • In this study, the effects of a pre-filter system and electrolysis system on the safe and efficient reuse of brine in the cabbage salting process were investigated. First, sediment filter-electrolyzed brine (SF-EB) was selected as brine for reuse. Then, we evaluated the quality and microbiological properties of SF-EB and Chinese cabbage salted with SF-EB. The salinity (9.4%) and pH (4.63) of SF-EB were similar to those of control brine (CB). SF-EB turbidity was decreased (from 0.112 to 0.062) and SF-EB residual chlorine (15.86 ppm) was higher than CB residual chlorine (0.31 ppm), and bacteria were not detected. Salinity (2.0%), pH (6.21), residual chlorine (0.39 ppm), chromaticity, hardness, and chewiness of cabbage salted with SF-EB were similar to those of cabbage salted with CB. The total bacterial count in cabbage salted with CB was increased as the number of reuses increased (from 6.55 to 8.30 log CFU/g), whereas bacteria in cabbage salted with SF-EB was decreased (from 6.55 to 5.21 log CFU/g). These results show that SF-EB improved the reusability of brine by removing contaminated materials and by sterilization.

Effects of Phloretin, Cytochalasin B, and D-Fructose on 2-deoxy-D-Glucose Transport of the Glucose Transport System Present in Spodoptera frugiperda Clone 21-AE Cells

  • Lee Chong-Kee
    • 대한의생명과학회지
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    • 제12권1호
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    • pp.17-22
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    • 2006
  • The baculovirus expression system is a powerful method for producing large amounts of the human erythrocyte-type glucose transport protein, heterologously. Characterization of the expressed protein is expected to show its ability to transport sugars directly. To achieve this, it is a prerequisite to know the properties of the endogenous sugar transport system in Spodoptera frugiperda Clone 21 (Sf21) cells, which are commonly employed as a host permissive cell line to support the baculovirus replication. The Sf21 cells can grow well on TC-100 medium that contains 0.1% D-glucose as the major carbon source, strongly suggesting the presence of endogenous glucose transport system. However, unlike the human glucose transport protein that has a broad substrate and inhibitor specificity, very little is known about the nature of the endogenous sugar transport system in Sf21 cells. In order to characterize further the inhibitor recognition properties of the Sf21 cell transporter, the ability of phloretin, cytochalasin B and D-fructose to inhibit 2-deoxy-D-glucose (2dGlc) transport was examined by measuring inhibition constants $(K_i)$. The $K_i's$ for reversible inhibitors were determined from plots of uptake versus inhibitor concentration. The 2dGlc transport in the Sf21 cells was very potently inhibited by phloretin, the aglucone of phlorizin with a $K_i$ similar to the value of about $2{\mu}M$ reported for inhibition of glucose transport in human erythrocytes. However, the Sf21 cell transport system was found to differ from the human transport protein in being much less sensitive to inhibition by cytochalasin B (apparent $K_i$ approximately $10\;{\mu}M$). In contrast, It is reported that the inhibitor binds the human erythrocyte counterpart with a $K_d$ of approximately $0.12\;{\mu}M$. Interestingly, the Sf21 glucose transport system also appeared to have high affinity for D-fructose with a $K_i$ of approximately 5mM, contrasting the reported $K_m$ of the human erythrocyte transport protein for the ketose of 1.5M.

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Comparative Characterization of Growth and Recombinant Protein Production among Three Insect Cell Lines with Four Kinds of Serum Free media

  • Kwon, Mi-Sun;Takashi Dojima;Park, Enoch Y.
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제8권2호
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    • pp.142-146
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    • 2003
  • Three insect cell lines, Sf9, Sf21 and Tn5Bl-4, and four different kinds of serum free media (SFM), Sf 900 II, EX-CELL 420, EX-CELL 405 and Express Five, were used to compare the nutrient consumption, byproduct formation, production of recombinant protein and protease activity in suspension cultures. The Sf 900 II SFM was a ppropriate for the cell growth and protein production of the Sf9 and Sf21 cell lines. When the Tn5Bl-4 cell line was grown in the Express Five SFM, the specific growth rate was 1.6 fold higher than those of either the Sf9 or Sf21 cell lines. The glucose and glutamine consumption rates per cells, were 4 and 2.3 times higher than those of the Sf9 cell line, respectively. The overall yield coefficients of the lactate and ammoniumion were 2.8 and 1.5 times higher compared to those of the Sf9 cell line. respectively. The maximum specific ${\beta}$-galactosidase production rate was 4.5 fold that of the Sf9 cell line, a 3 times higher protease activity per cell.

Hexose Uptake and Kinetic Properties of the Endogenous Sugar Transporter(s) in Spodoptera frugiperda Clone 21-AE Cells

  • Lee Chong-Kee
    • 대한의생명과학회지
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    • 제11권3호
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    • pp.327-332
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    • 2005
  • Sf21 cells become popular as the host permissive cell line to support the baculovirus AcNPV replication and protein synthesis. The cells grow well on TC-100 medium that contains $0.1\%$ D-glucose as the major carbon source, strongly suggesting the presence of endogenous glucose transporters. However, unlike human glucose transporters, very little is known about the characteristics of the endogenoussugar transporter(s) in Sf21 cells. Thus, some kinetic properties of the sugar transport system were investigated, involving the uptake of 2-deoxy-D-glucose (2dG1c). In order to obtain a true measure of the initial rate of uptake, the uptake of $[^3H]2dGlc$ from both low $(100{\mu}M)$ and high (10 mM) extracellular concentrations was measured over periods ranging from 30 sec to30 min. The data obtained indicated that the uptake was linear for at least 2 min at both concentrations, suggesting that measurements made over a 1min time course would reflect initial rates of the jexpse uptake. To determine $K_m\;and\;V_{max}$ of the endogenous glucose transporter(s) in Sf21 cells, the uptake of 2dG1c was measured over a range of substrate concentrations $(50{\mu}M\~10mM)$ 2dG1c uptake by the Sf21 cells appeared to involve both saturable and non-saturable (or very low affinity) components. A saturable transport system for 2dG1c was relatively high, the $K_m$ value for uptake being < 0.45 mM. The $V_{max}$ value obtained for 2dG1c transport in the Sf21 cells was about 9.7-folds higher than that reported for Chinese hamster ovary cells, which contain a GLUT1 homologue. Thus, it appeared that the transport activity of the Sf21 cells was very high. In addition, the Sf21 glucose transporter was found to have very low affinity for cytochalasin B, a potent inhibitor of human erythrocyte glucose transporter

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한국 SF의 장르적 특징과 의의 -근대화에 대한 프로파간다부터 포스트휴먼 담론까지 (Characteristics and Meanings of the SF Genre in Korea - From Propaganda of Modernization to Post-Human Discourse)

  • 이지용
    • 대중서사연구
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    • 제25권2호
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    • pp.33-69
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    • 2019
  • 본 고는 한국에서 SF가 장르적으로 어떠한 의미를 지니고 있는지에 대해 밝히고자 하는 목적을 가지고 있다. 이제까지 한국에서 SF가 가지고 있었던 특징에 대한 연구는 주로 SF가 가지고 있는 특징적인 요소들에 대한 의미를 밝히거나 작품에 대한 특징들을 지엽적으로 리뷰하는 형식이 주를 이루었다. 하지만 이러한 방법론은 장르로서의 특징을 가지고 있는 SF 텍스트들에 대한 정체성을 간과하고 기존의 방법론을 통해 분석했다는 한계가 있다. SF라는 장르 텍스트의 가치를 명확하게 규정하기 위해서는 장르의 관습과 코드에 대한 이해가 선결되어야 하기 때문이다. 그러기 때문에 본 고는 우선 한국의 SF가 어떤 역사적 맥락을 가지고 들어와서 한국 사회에 수용됐으며, 그 안에서 어떤 의미와 특징들을 가지고 창작되고 독자들과 관계를 맺었는지에 대해 개괄적으로 다루고자 한다. 이를 통해 그동안 학술담론에서 본격적으로 다루지 않았던 대중 서사 및 장르서사로서의 SF에 대해 본격적으로 조망하고, 21세기를 지나 과학적 상상력이 중요해진 시대에 각광을 받으면서 사회적으로 재조명 되기 시작한 SF가 가지고 있는 현재와 미래의 가능성에 대해 실질적으로 재고해 보고자 한다. 본 고는 한국의 SF 텍스트들이 학술담론 내에서 의미작용을 하는데 기본이 되는 배경을 위한 것이다. 이 작업을 통해 그동안 문학의 영역에서도, 그리고 문화현상의 영역에서도 제대로 다루어지지 않았던 수많은 한국 SF는 이후로 학술담론 내에서 유의미한 가치들을 평가받고 이후로 다양한 연구를 통해 재조명될 것이다. 그리고 이러한 작업을 통해 결과적으로 21세기 이후에 다양하게 변주되어 나타나고 있는 한국 서사 영역의 확장과 담론의 발달에도 도움이 될 수 있을 것이라 기대한다.

A Productive Replication of Hyphantria cunea Nucleopolyhedrovirus in Lymantria dispar Cell Line

  • Demir, Ismail;Demirbag, Zihni
    • Journal of Microbiology and Biotechnology
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    • 제16권10호
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    • pp.1485-1490
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    • 2006
  • In this study, comparative replicational properties of Hyphantria cunea nucleopolyhedrovirus (HycuNPV) in Lymantria dispar (IPLB-LdElta) and Spodoptera frugiperda (IPLB-Sf21) cell lines were investigated. Our microscopic observations showed that cytopathic effects (CPEs) in LdElta cells appeared 12 h later than those in Sf21 cells. Whereas polyhedral inclusion bodies (PIBs) formed at 48 h postinfection (p.i.) in LdElta cells, it formed at 36 h p.i. in Sf21 cells. Extracellular virus production determined according to the 50% tissue culture infective dose ($TCID_{50}$) method in LdElta cells started about 12 h later when compared with Sf21 cells. Titers of extracellular virus in LdElta and Sf21 cells were calculated as $1.77{\times}10^9$ plaque forming units (PFU)/ml and $5.6{\times}10^9PFU/ml$, respectively, at 72 h p.i. We also showed that viral DNA replication began at 12 h p.i. in both cell lines. Viral protein synthesis was determined by SDS-polyacrylamide gel electrophoresis (PAGE) and polyhedrin synthesis was observed at 12 h p.i. in both cell lines. The results indicate that while the synthesis of macromolecules is 12 h later and production of extracellular virus is almost 3-fold lower in LdElta cells compared with those in Sf21 cells, the LdElta cell line is still a productive cell line for infection of HycuNPV.

The geometry change of carbon nanofilaments by SF6 incorporation in a thermal chemical vapor deposition system

  • Kim, Sung-Hoon
    • 한국결정성장학회지
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    • 제21권3호
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    • pp.119-123
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    • 2011
  • Carbon nanotilaments (CNFs) could be synthesized on nickel catalyst layer-deposited silicon oxide substrate using $C_2H_2$ and$H_2$ as source gases under thermal chemical vapor deposition system. By the incorporation of $SF_6$ as a cyclic modulation manner, the geometries of carbon coils-related materials, such as nano-sized coil and wave-like nano-sized coil could be observed on the substrate. The characteristics (formation density and morphology) of as-grown CNFs with or without $SF_6$ incorporation were investigated. Diameter size reduction for the individual CNFs-related shape and the enhancement of the formation density of CNFs-related material could be achieved by the incorporation of $SF_6$ as a cyclic modulation manner. The cause for these results was discussed in association with the slightly increased etching ability by $SF_6$ addition and the sulfur role in SF 6 for the geometry change.

Propagation of Bombyx mori Nucleopolyhedrovirus in Nonpermissive Insect Cell Lines

  • Woo, Soo-Dong;Roh, Jong-Yul;Choi, Jae-Young;Jin, Byung-Rae
    • Journal of Microbiology
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    • 제45권2호
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    • pp.133-138
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    • 2007
  • This study addresses the susceptibility of Spodoptera frugiperda (Sf9 and Sf21), Trichoplusia ni (Hi5), and S. exigua (Se301) cells to the Bombyx mori nucleopolyhedrovirus (BmNPV). Although these cells have classically been considered nonpermissive to BmNPV, the cytopathic effect, an increase in viral yield, and viral DNA synthesis by BmNPV were observed in Sf9, Sf21, and Hi5 cells, but not in Se301 cells. Very late gene expression by BmNPV in these cell lines was also detected via ${\beta}-galactosidase$ expression under the control of the polyhedrin promoter. Sf9 cells were most susceptible to BmNPV in all respects, followed by Sf21 and Hi5 cells in decreasing order, while the Se301 cells evidenced no distinct viral replication. This particular difference in viral susceptibility in each of the cell lines can be utilized for our understanding of the mechanisms underlying the host specificity of NPVs.

파밤나방 세포주에서 Autographa californica 핵다각체병 바이러스의 감염 특성 (Characteristics of Autographa californica Nuclear Polyhedrosis Virus in Spodoptera exigua Cell Line.)

  • 최재영;우수동;홍혜경;강석권
    • 한국미생물·생명공학회지
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    • 제26권2호
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    • pp.161-166
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    • 1998
  • AcNPV에 대해 감수성을 보이는 파밤나방 세포주인 Se301 세포주의 AcNPV를 이용한 발현 벡터계에서의 유용성을 알아보기 위하여 Se301세포주에서 AcNPV의 감염 특성을 Sf-21 세포주에서와 비교하였다. 그 결과, 바이러스의 감염상은 두 세포주에서 거의 유사하였으나 감염 7일 후에 세포로부터 방출된 다각체의 비율은 Se301 세포주에서 훨씬 높았다. 또한 전체 다각체 생성율은 Se301 세포주에서 약간 높았지만 그 차이는 매우 근소하여 대체적으로 두 세포주에서 거의 비슷하였으나 생성된 다각체의 크기는 Se301세포주에서 더욱 컸다. 한편, 세포주나 바이러스의 증식율과는 무관하게 AcNPV에 의한 다각체 단백질의 발현량이 Se301 세포주에서 약 2.4배 높게 나타남으로써 Se301 세포주가 AcNPV를 이용한 발현 벡터계에서 숙주 세포로서 매우 유용하게 이용될 수 있음을 보여 주었다.

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