• Title/Summary/Keyword: Sf cell

Search Result 192, Processing Time 0.035 seconds

SF3B4 as an early-stage diagnostic marker and driver of hepatocellular carcinoma

  • Shen, Qingyu;Nam, Suk Woo
    • BMB Reports
    • /
    • v.51 no.2
    • /
    • pp.57-58
    • /
    • 2018
  • An accurate diagnostic marker for detecting early-stage hepatocellular carcinoma (eHCC) is clinically important, since early detection of HCC remarkably improves patient survival. From the integrative analysis of the transcriptome and clinicopathologic data of human multi-stage HCC tissues, we were able to identify barrier-to-autointegration factor 1 (BANF1), procollagen-lysine, 2-oxoglutarate 5-dioxygenase 3 (PLOD3) and splicing factor 3b subunit 4 (SF3B4) as early HCC biomarkers which could be detected in precancerous lesions of HCC, with superior capabilities to diagnose eHCC compared to the currently popular HCC diagnostic biomarkers: GPC3, GS, and HSP70. We then showed that SF3B4 knockdown caused G1/S cell cycle arrest by recovering $p27^{kip1}$ and simultaneously suppressing cyclins, and CDKs in liver cancer cells. Notably, we demonstrated that aberrant SF3B4 overexpression altered the progress of splicing progress of the tumor suppressor gene, kruppel like factor 4 (KLF4), and resulted in non-functional skipped exon transcripts. This contributes to liver tumorigenesis via transcriptional inactivation of $p27^{kip1}$ and simultaneous activation of Slug genes. Our results suggest that SF3B4 indicates early-stage HCC in precancerous lesions, and also functions as an early-stage driver in the development of liver cancer.

Functional Properties of Human Muscarinic Receptors Hm1, Hm2 and Hm3 Expressed in a Baculovirus/Sf9 Cell System

  • Woo, Hyun-Ae;Woo, Yae-Bong;Bae, Seung-Jin;Kim, Hwa-Jung
    • Biomolecules & Therapeutics
    • /
    • v.7 no.4
    • /
    • pp.307-314
    • /
    • 1999
  • The human muscarinic acetylcholine receptor (mAChR) subtypes Hml, Hm2 and Hm3 have been expressed in insect cells (Spodoptera frugiperda, Sf9) using the baculovirus expression system. Expression of relevant DNA, transcript and receptor proteins was identified by PCR, Northern blotting and [$^{3}H$]QNB binding, respectively. As assessed by [$^{3}H$]QNB binding sites, yields of muscarinic receptors in membrane preparations in this study were as about 5-20 times high as those in mammalian cells reported in previous studies. The [$^{3}H$]QNB competition binding studies with well-known subtype-selective mAChR antagonists showed that the receptors expressed in Sf9 cells retain the pharmacological characteristics expected for the ml , m2 and m3 muscarinic receptors. The ml-selective antagonist, pirenzepine, displayed a considerably higher affinity for Hml by 110-fold and 35-fold than for Hm2 and Hm3, respectively, The m2-selective methoctramine displayed a significantly higher affinity for Hm2 than for Hml and Hm3 (10- and 26-fold, respectively). p-F-HHSiD exhibited high affinity for Hm3 that is not significantly different from those for Hml, but 66-fold higher than its affinity for Hm2. The functional coupling of the recombinant receptors to second messenger systems was also examined. While both Hml and Hm3 stimulated phosphoinositide hydrolysis upon activation by carba-chol, Hm2 produced no response. On the other hand, activation of mAChRs induced the inhibition of forsko-lin-stimulated cyclic AMP formation in Hm2-expressing cells, whereas the significant dose-dependent increase in or poor response on cyclic AMP formation were produced in Hml or Hm3-expressing cells, respectively. These results indicate the differential coupling of recombinant Hml, Hm2 and Hm3 receptors expressed in SF9 cells to intracellular signalling system.

  • PDF

Relationship between Radiosensitivity and Repair Capacity in Human Epithelial Cancer Cell Lines (인체 상피암 세포주에서 방사선감수성과 손상회복의 상관관계에 관한 연구)

  • Koh, Kyoung-Hwan;Ha, Sung-Whan;Park, Charn-Il
    • Radiation Oncology Journal
    • /
    • v.11 no.1
    • /
    • pp.17-27
    • /
    • 1993
  • To investigate the relationship between radiosensitivity and postirradiation recovery in human cancer cells, a study was performed using human cancer cell lines-A549, CaSki, SNU-C5 and PCI-13. For the study of radiosensitivity, single doses of 2, 4, 6, 8, 10, 12, and 14 Gy were given and for postirradiation recovery, two fractions of 4 Gy were separated with a time interval of 0, 0.5, 1, 1.5, 2, 2.5, 3, 4, 5, or 6 hours. Surviving fraction was estimated using colony forming ability. Surviving fractions at 2 Gy (SF2) were 0.496 (0.570-0.412) for A549, 0.496 (0.660-0.332) for CaSki, 0.386 (0.576-0.216) for SNU-C5, and 0.185 (0.247-0.123) for PCI-13. By statistical analysis the SF2 of PCI-13 was lower significantly than those of others (p<0.05). This difference was also observed at 4, 6 and 8 Gy dose levels. At 6 and 8 Gy the surviving fractions of SNU-C5 were also lower significantly than A549 and CaSki (p<0.05). By the analysis with linear quadratic model, the values of ${\alpha}$ for A549, CaSki, SNU-C5 and PCI-13 were 0.3016, 0.3212, 0.4327 and 0.8423, respectively, and those of ${\beta}$ were 0.02429, 0.02009, 0.03349 and 0.00059, respectively. So, the value of ${\alpha}$ showed increasing tendency with decreasing SF2. By the multitarget single hit model the values of Do for A549, CaSki, SUN-C5 and PCI-13 were 1.97, 1.97, 1.46 and 0.81, respectively, and those of n were 1.53, 1.50, 1.56 and 2.28, respectively. So, the value of Do decreased with decreasing SF2. Post-irradiation recovery reached plateau at around 2 hours. Recovery ratio at plateau phase ranged from 1.2 to 4.2; the value were 1.2 for PCI-13, 3.2 for CaSki, 3.3 for SNU-C5, and 4.2 for A549. Recovery rate well correlated with SF2, and increased with increasing Do and decreasing ${\alpha}$. According to above results, the intrinsic radiosensitivity was quite different among the tested cell lines; PCI-13 was the most sensitive and A549 and CaSki was similar. This difference of radiosensitivity is thought to be partly due to the difference in amount of postirradiation recovery. By linear quadratic model the difference of ${\alpha}$ values was very high, and by multitarget single hit model the difference of Do value was significantly high among four cell lines.

  • PDF

Study on Spectrum of Partial Discharge under Air, $SF_6$ and Oil (기중, $SF_6$ 및 절연유 하에서의 방전신호 주파수 특성분석)

  • Goo, Sun-Geun;Ju, Hyung-Jun;Han, Ki-Seon;Park, Ki-Jun;Yoon, Jin-Yul
    • Proceedings of the KIEE Conference
    • /
    • 2006.10a
    • /
    • pp.149-150
    • /
    • 2006
  • 기중, $SF_6$ 가스 및 절 연유에서 부분방전시 방사되는 전자파의 주파수 특성을 분석하였다. 비슷한 구성의 부분방전 발생장치(PD Cell)에서 절연유에서의 부분방전이 가장 최고 높은 주파수의 신호가 발생하였으며 $SF_6$ 가스 하에서 비교적 높은 주파수의 부분방전 신호가 발생하였다. 또한 부분 방전의 종류에 따라 전자파 방전신호가 수직 편파되어 있음을 알 수 있었나. 본 결과는 전자파를 이용한 변전기기의 부분방전 측정기술 개발에 응용할 예정이다.

  • PDF

The Molecular Biological Marker in Bombyx mori and Spodoptera frugiperda Cells (Bombyx mori세포주와 Spodoptera frugiperda세포주의 분자생물학적 표식자)

  • Jin, Byeong-Rae;Je, Yeon-Ho;Gang, Seok-Gwon
    • Journal of Sericultural and Entomological Science
    • /
    • v.38 no.1
    • /
    • pp.53-56
    • /
    • 1996
  • To investigate the molecular biological marker in insect cells, BmN-4 and Sf-0 cells were analysed by SDS-PAGE and random amplification of polymorphic DNA. The results showed that the patterns of total cell protein and random amplification of polymorphic DNA were distinguished between BmN-4 and Sf-9 cells, suggesting that the unique major bands were useful as molecular biological marker in BmN-4 and Sf-9 cells.

  • PDF

Efficient Symbol Detection Algorithm for Space-frequency OFDM Transmit Diversity Scheme (공간-주파수 OFDM 전송 다이버시티 기법을 위한 효율적인 심볼 검출 알고리즘)

  • Jung Yun ho;Kim Jae seok
    • The Journal of Korean Institute of Communications and Information Sciences
    • /
    • v.30 no.4C
    • /
    • pp.283-289
    • /
    • 2005
  • In this paper, we propose two efficient symbol detection algorithms for space-frequency OFDM (SF-OFDM) transmit diversity scheme. When the number of sub-carriers in SF-OFBM scheme is small, the interference between adjacent sub-carriers may be generated. The proposed algorithms eliminate this interference in a parallel or sequential manlier and achieve a considerable performance improvement over the conventional detection algorithm. The bit error rate (BER) performance of the proposed detection algorithms is evaluated by the simulation. In the case of 2 transmit and 2 receive antennas, at $BER=10^{-4}$ the proposed algorithms achieve the gain improvement of about 3 dB. The symbol detectors with the proposed algorithms are designed in a hardware description language and synthesized to gate-level circuits with the $0.18{\mu}m$ 1.8V CMOS standard cell library. With the division-free architecture, the proposed SF-OFDM-PIC and SF-OFDM-SIC symbol detectors can be implemented using 140k and 129k logic gates, respectively.

Anti-inflammatory activity of AP-SF, a ginsenoside-enriched fraction, from Korean ginseng

  • Baek, Kwang-Soo;Hong, Yong Deog;Kim, Yong;Sung, Nak Yoon;Yang, Sungjae;Lee, Kyoung Min;Park, Joo Yong;Park, Jun Seong;Rho, Ho Sik;Shin, Song Seok;Cho, Jae Youl
    • Journal of Ginseng Research
    • /
    • v.39 no.2
    • /
    • pp.155-161
    • /
    • 2015
  • Background: Korean ginseng is an ethnopharmacologically valuable herbal plant with various biological properties including anticancer, antiatherosclerosis, antidiabetic, and anti-inflammatory activities. Since there is currently no drug or therapeutic remedy derived from Korean ginseng, we developed a ginsenoside-enriched fraction (AP-SF) for prevention of various inflammatory symptoms. Methods: The anti-inflammatory efficacy of AP-SF was tested under in vitro inflammatory conditions including nitric oxide (NO) production and inflammatory gene expression. The molecular events of inflammatory responses were explored by immunoblot analysis. Results: AP-SF led to a significant suppression of NO production compared with a conventional Korean ginseng saponin fraction, induced by both lipopolysaccharide and zymosan A. Interestingly, AP-SF strongly downregulated the mRNA levels of genes for inducible NO synthase, tumor necrosis factor-${\alpha}$, and cyclooxygenase) without affecting cell viability. In agreement with these observations, AP-SF blocked the nuclear translocation of c-Jun at 2 h and also reduced phosphorylation of p38, c-Jun N-terminal kinase, and TAK-1, all of which are important for c-Jun translocation. Conclusion: Our results suggest that AP-SF inhibits activation of c-Jun-dependent inflammatory events. Thus, AP-SF may be useful as a novel anti-inflammatory remedy.

Plant Regeneration from Protoplasts of Suspension Cultured Cells in Arabidopsis thaliana (애기장대(Arabidopsis thaliana) 현탁배양세포의 원형질체로부터 식물체 재분화)

  • 김명덕;김준철;진창덕;임창진;한태진
    • Korean Journal of Plant Tissue Culture
    • /
    • v.27 no.2
    • /
    • pp.125-131
    • /
    • 2000
  • Protoplasts of Arabidopsis thaliana were easily isolated from the shoot-forming (SF) suspension-cultured cell clusters with 4 hours-shaking condition (40 rpm) on CPD enzyme solution containing 1% cellulase R-10, 0.25% pectolyase Y-23 and 0.5% driselase. Protoplasts were cultured on liquid KAO medium supplemented with 1 mg/L 2,4-D, 0.5 mg/L kinetin, 200 mg/L spermidine and 68 g/L glucose. Also, protoplasts were cultured on 0.2 $\mu$M membrane filter placed onto CP solid medium containing the suspension cells as feeder cells in the dark at $25^{\circ}C$ for 4 weeks. Protoplast-derived-SF calli were cultured on MS medium containing 0.05 mg/L IAA, 7 mg/L 2 ip and 30 g/L sucrose under the continuous illumination for four weeks. The frequency of shoot formation was about 60%. The regenerants were transferred into potting soil to grow mature plants. The regenerants formed the silques with seeds after 8 weeks of cultures.

  • PDF

The Modulation of Radiosensitivity by Combined Treatment of Selective COX-2 Inhibitor, NS 398 and EGF Receptor Blocker AG 1478 in HeLa Cell Line (선택적 COX-2 억제제 NS 398과 EGF 수용체 차단제 AG 1478의 복합투여가 HeLa 세포주의 방사선 감수성에 미치는 영향)

  • Youn Seon Min;Oh Young Kee;Kim Joo Heon;Park Mi Ja;Seong In Ock;Kang Kimun;Chai Gyuyong
    • Radiation Oncology Journal
    • /
    • v.23 no.1
    • /
    • pp.51-60
    • /
    • 2005
  • Purpose : Selective inhibition of multiple molecular targets may improve the antitumor activity of radiation. Two specific inhibitors of selective cyclooxygenase-2 (COX-2) and epidermal growth factor receptor (EGFR) were combined with radiation on the HeLa cell line. To investigate cooperative mechanism with selective COX-2 inhibitor and EGFR blocker, in vitro experiments were done. Materials and Methods : Antitumor effect was obtained by growth inhibition and apoptosis analysis by annexin V-Flous method. Radiation modulation effects were determined by the clonogenic cell survival assay. Surviving fractions at 2 Gy ($SF_2$) and dose enhancement ratio at a surviving fraction of 0.25 were evaluated. To investigate the mechanism of the modulation of radiosensitivity, the cell cycle analyses were done by flow cytometry. The bcl-2 and bax expressions were analyzed by western blot. Results : A cooperative effect were observed on the apoptosis of the HeLa ceil line when combination of the two drugs, AG 1478 and NS 398 with radiation at the lowest doses, apoptosis of $22.70\%$ compare with combination of the one drug with radiation, apoptosis of $8.49\%$. In cell cycle analysis, accumulation of cell on $G_0/G_l$ phase and decrement of S phase fraction was observed from 24 hours to 72 hours after treatment with radiation, AG 1478 and NS 398. The combination of NS 398 and AG 1478 enhanced radiosensitivity on a concentration-dependent manner in HeLa cells with dose enhancement ratios of 3.00 and $SF_2$ of 0.12 but the combination of one drug with radiation was not enhanced radlosensitivity with dose enhancement ratios of 1.12 and SF2 of 0.68 (p=0.005). The expression levels of bcl-2 and bax were reduced when combined with AG 1478 and NS 398. Conclusion : Our results indicate that the selective COX-2 inhibitor and EGFR blocker combined with radiation have potential additive or cooperative effects on radiation treatment and may act through various mechanisms including direct inhibition of tumor cell proliferation, suppression of tumor cell cycle progression and inhibition of anti-apoptotic proteins.

Large-Scale Production of Rotavirus VLP as Vaccine Candidate Using Baculovirus Expression Vector System (BEVS)

  • Park, Jin-Yong;Kim, Hun;Hwang, Hi-Ku;Lee, Su-Jeen;Kim, Hyun-Sung;Hur, Byung-Ki;Ryu, Yeon-Woo;An, Chang-Nam;Kim, Jong-Soo
    • Journal of Microbiology and Biotechnology
    • /
    • v.14 no.1
    • /
    • pp.35-40
    • /
    • 2004
  • Rotavirus virus-like particle (VLP) composed of VP2, VP6, and VP7 was expressed in the Baculovirus Expression Vector System (BEVS). Sf9 cell, a host of the baculovirus, was cultured from a 0.5-1 spinner flask to the 50-1 bioreactor system. Sf9 cell was maintained at cell density between 3.0E+05 and 3.0E+06 cells/ml and grew up to 1.12E+07 cells/ml in the bioreactor. Growth kinetics was compared under different culture systems and showed similar growth kinetics with 20.1-25.2 h of doubling time. Early exponentially growing cell culture was infected with three recombinant baculoviruses expressing VP2, VP6, and VP7 protein at 1.0, 2.0, and 0.2 moi, respectively. The expression of rotavirus proteins was confirmed by Western blot analysis and its three-layered virus-like structure was observed under an electron microscope. Rotavirus VLP was semipurified and immunized in ICR mice intramuscularly. Rotavirus-specific serum antibody was detected from 2 weeks after the immunization and lasted at least 21 weeks of the post-immunization, indicating its possible use as a vaccine candidate.