• Title/Summary/Keyword: cell cycling

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RNAi-based Knockdown of Multidrug Resistance-associated Protein 1 is Sufficient to Reverse Multidrug Resistance of Human Lung Cells

  • Shao, Shu-Li;Cui, Ting-Ting;Zhao, Wei;Zhang, Wei-Wei;Xie, Zhen-Li;Wang, Chang-He;Jia, Hong-Shuang;Liu, Qian
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.24
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    • pp.10597-10601
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    • 2015
  • Up-regulation of multidrug resistance-associated protein 1 (MRP1) is regarded as one of the main causes for multidrug resistance (MDR) of tumor cells, leading to failure of chemotherapy-based treatment for a multitude of cancers. However, whether silencing the overexpressed MRP1 is sufficient to reverse MDR has yet to be validated. This study demonstrated that RNAi-based knockdown of MRP1 reversed the increased efflux ability and MDR efficiently. Two different short haipin RNAs (shRNAs) targeting MRP1 were designed and inserted into pSilence-2.1-neo. The shRNA recombinant plasmids were transfected into cis-dichlorodiamineplatinum-resistant A549 lung (A549/DDP) cells, and then shRNA expressing cell clones were collected and maintained. Real time PCR and immunofluorescence staining for MRP1 revealed a high silent efficiency of these two shRNAs. Functionally, shRNA-expressing cells showed increased rhodamine 123 retention in A549/DDP cells, indicating reduced efflux ability of tumor cells in the absence of MRP1. Consistently, MRP1-silent cells exhibited decreased resistance to 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) and DDP, suggesting reversal of MDR in these tumor cells. Specifically, MRP1 knockdown increased the DDP-induced apoptosis of A549/DDP cells by increased trapping of their cell cycling in the G2 stage. Taken together, this study demonstrated that RNAi-based silencing of MRP1 is sufficient to reverse MDR in tumor cells, shedding light on possible novel clinical treatment of cancers.

Study on Nucleo-Cytoplasmic Interaction by Somatic Cell Nuclear Transfer in Bovine (소 체내포 핵이식에 의한 핵-세포질 상호작용에 관한 연구)

  • 정희태;최종엽;박춘근;김정익;민동미
    • Journal of Embryo Transfer
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    • v.15 no.1
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    • pp.23-31
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    • 2000
  • This study was conducted to investigate the effects of quiescent treatment of donor cells and activation treatment time of recipient cytoplasm on nuclear remodeling and in vitro development of somatic cell-cloned bovine embryos. Serum starved, confluent and nonquiescent cycling adult skin cells were teansferred into enucleated oocytes. Nuclear transfer oocytes were activated at 30 min, 1 and 2 hrs after electrofusion. Some nuclear transfer embryos(23% to 35%) extruded a polar body, which was not affected by quiescent treatment of donor cells and activiation time of recipient cytoplasm. About 68% of nuclear transfer embryos fused with a serum starved cells has a chromatin clump, but which was not different from embryos fused with confluent(51%) and nonquiescent(47%) cells. The proportion of embryos with a single chromatin clump was sightly increased when nuclear transfer embryos were activated within 30 min after fusion(69%) compared to those were activated at 1 and 2 hrs after fusion, but there was not significantly different. Development rates to the blastocyst stage were 8.6% and 15.9% when serum starved and confluent cells were transferred, which were higher than that of control group. Developmental rate to the blastocyst stage was higher in embryos were activated within 30 min after fusion (17.3%) compared to those of embryos were activated at 1 and 2 hrs after fusion (P<0.05). From the present result, it is suggested that quiescent treatment of donor cells and activation time of recipient cytoplasm can affect the in vitro development. Quiescent plasm activation within 30 min after fusion could increase the number of embryos with a normal chromation structure, which results in increased in vitro development.

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Surface Modification of Synthetic Graphite as an Electrode by Fluidized-bed Chemical Vapor Deposition for Lithium Secondary Batteries (유동상 화힉증착에 의한 리튬이차전지 전극용 탄소재료의 표면개질)

  • Ryu D. H.;Lee Joong Kee;Park D. G.;Yun K. S.;Cho B. W.;Shul Y. G.
    • Journal of the Korean Electrochemical Society
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    • v.3 no.3
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    • pp.173-177
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    • 2000
  • The synthetic carbon was coated with tin oxide and copper by fluidized-bed chemical vapor deposition method. $(CH_3)_4Sn\;and\;Cu(hfac)_{2s}$ were employed as the metallic organic precursor, respectively. The modified synthetic carbons were used for lithium secondary battery anode to investigate their coating effects on electrochemical characteristics as alternative anode materials for lithium secondary batteries. The electrode which prepared by the synthetic carbons(MCMB) coated with tin oxide gave the higher capacity than that of raw material. Their capacity decreased with the progress of cycling possibly due to severe volume changes. But the cyclability was improved by coating with copper on the surface of the tin oxide coated carbon, which plays an important role as an inactive matrix buffering volume changes.

Effects of Nickel and Iron Oxide Addition by Milling under Hydrogen on the Hydrogen-Storage Characteristics of Mg-Based Alloys

  • Song, Myoung Youp;Baek, Sung Hwan;Park, Hye Ryoung;Mumm, Daniel R.
    • Korean Journal of Metals and Materials
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    • v.50 no.1
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    • pp.64-70
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    • 2012
  • Samples of pure Mg, 76.5 wt%Mg-23.5 wt%Ni, and 71.5 wt%Mg-23.5 wt%Ni-5 wt%$Fe_2O_3$ were prepared by reactive mechanical grinding and their hydriding and dehydriding properties were then investigated. The reactive mechanical grinding of Mg with Ni is considered to facilitate nucleation and to shorten diffusion distances of hydrogen atoms. After hydriding-dehydriding cycling, the 76.5 wt%Mg-23.5 wt%Ni and 71.5 wt%Mg-23.5 wt%Ni-5 wt%$Fe_2O_3$ samples contained $Mg_2Ni$ phase. In addition to the effects of the creation of defects and the decrease in particle size, the addition of Ni increases the hydriding and dehydriding rates by the formation of $Mg_2Ni$. Expansion and contraction of the hydride-forming materials (Mg and $Mg_2Ni$) with the hydriding and dehydriding reactions are also considered to increase the hydriding and dehydriding rates of the mixture by forming defects and cracks leading to the fragmentation of particles. The reactive mechanical grinding of Mg-Ni alloy with $Fe_2O_3$ is considered to decrease the particle size.

Suppression of Protein Kinase C and Nuclear Oncogene Expression as Possible Action Mechanisms of Cancer Chemoprevention by Curcumin

  • Lin, Jen-Kun
    • Archives of Pharmacal Research
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    • v.27 no.7
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    • pp.683-692
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    • 2004
  • Curcumin (diferuloylmethane) is a major naturally-occurring polyphenol of Curcuma species, which is commonly used as a yellow coloring and flavoring agent in foods. Curcumin has shown anti-carcinogenic activity in animal models. Curcumin possesses anti-inflammatory activity and is a potent inhibitor of reactive oxygen-generating enzymes such as lipoxygenase/cyclooxygenase, xanthine dehydrogenase/oxidase and inducible nitric oxide synthase; and an effective inducer of heme oxygenase-1. Curcumin is also a potent inhibitor of protein kinase C(PKC), EGF(Epidermal growth factor)-receptor tyrosine kinase and LĸB kinase. Subsequently, curcumin inhibits the activation of NF(nucleor factor)KB and the expressions of oncogenes including c-jun, c-fos, c-myc, NIK, MAPKs, ERK, ELK, PI3K, Akt, CDKs and iNOS. It is proposed that curcumin may suppress tumor promotion through blocking signal transduction path-ways in the target cells. The oxidant tumor promoter TPA activates PKC by reacting with zinc thiolates present within the regulatory domain, while the oxidized form of cancer chemopreventive agent such as curcumin can inactivate PKC by oxidizing the vicinal thiols present within the catalytic domain. Recent studies indicated that proteasome-mediated degradation of cell proteins playa pivotal role in the regulation of several basic cellular processes including differentiation, proliferation, cell cycling, and apoptosis. It has been demonstrated that curcumin-induced apoptosis is mediated through the impairment of ubiquitin-proteasome pathway. Curcumin was first biotransformed to dihydrocurcumin and tetrahydrocurcumin and that these compounds subsequently were converted to monoglucuronide conjugates. These results suggest that curcumin-glucuronide, dihydrocurcumin-glucuronide, tetrahydrocurcumin-glucuronide and tetrahydrocurcumin are the major metabolites of curcumin in mice, rats and humans.

Surface Morphology Changes of Lithium/Sulfur Battery using Multi-walled carbon nanotube added Sulfur Electrode during Cyclings (탄소나노튜브가 첨가된 유황전극을 사용한 리튬/유황 전지의 사이클링에 의한 표면형상변화)

  • Park, Jin-Woo;Yu, Ji-Hyun;Kim, Ki-Won;Ryu, Ho-Suk;Ahn, Jou-Hyeon;Jin, Chang-Soo;Shin, Kyung-Hee;Kim, Young-Chul;Ahn, Hyo-Jun
    • Korean Journal of Metals and Materials
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    • v.49 no.2
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    • pp.174-179
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    • 2011
  • We investigated the surface morphology changes of a lithium/sulfur battery using multi-walled canbon nanotube added sulfur electrode during charge-discharge cycling. The Li/S cell showed the first discharge capacity of 1286 mAh/g-S, which utilized is 71% of the theoretical value. It decreased to 328 mAh/g-S at the 100th cycle, which corresponds to about 19% utilization of the total sulfur in the cathode. The spherical lumps of the reaction product were observed on the surface of the sulfur electrode. This material was verified as lithium sulfide by X-ray diffraction measurement. The pores in the separator were filled with reaction product. Thus the diffusion of the $Li^+$ ion decreased, which resulted in the decreased capacity of the Li/S cell.

Y2O3-stabilized ZrO2, Ni, and graphene-added Mg by reactive mechanical grinding processing for hydrogen storage and comparison with Ni and Fe2O3 or MnO-added Mg

  • Song, Myoung Youp;Choi, Eunho;Kwak, Young Jun
    • Journal of Ceramic Processing Research
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    • v.20 no.6
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    • pp.609-616
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    • 2019
  • The optimum powder to ball ratio was examined, which is one of the important conditions in reactive mechanical grinding processing. Yttria (Y2O3)-stabilized zirconia (ZrO2) (YSZ), Ni, and graphene were chosen as additives to enhance the hydriding and dehydriding rates of Mg. Samples with a composition of 92.5 wt% Mg + 2.5 wt% YSZ + 2.5 wt% Ni + 2.5 wt% graphene (designated as Mg-2.5YSZ-2.5Ni-2.5graphene) were prepared by grinding in hydrogen atmosphere. Mg-2.5YSZ-2.5Ni-2.5graphene had a high effective hydrogen-storage capacity of almost 7 wt% (6.85 wt%) at 623 K in 12 bar H2 at the second cycle (n = 2). Mg-2.5YSZ-2.5Ni-2.5graphene contained Mg2Ni phase after hydriding-dehydriding cycling. Mg-2.5YSZ-2.5Ni-2.5graphene had a larger quantity of hydrogen absorbed for 60 min, Ha (60 min), than Mg-2.5Ni-2.5graphene and Mg-2.5graphene. The addition of YSZ also increased the initial dehydriding rate and the quantity of hydrogen released for 60 min, Hd (60 min), compared with those of Mg-2.5Ni-2.5graphene. Y2O3-stabilized ZrO2, Ni, and graphene-added Mg had a higher initial hydriding rate and a larger Ha (60 min) than Fe2O3, MnO, or Ni and Fe2O3-added Mg at n = 1.

UV-cured Polymer Solid Electrolyte Reinforced using a Ceramic-Polymer Composite Layer for Stable Solid-State Li Metal Batteries

  • Hye Min Choi;Su Jin Jun;Jinhong Lee;Myung-Hyun Ryu;Hyeyoung Shin;Kyu-Nam Jung
    • Journal of Electrochemical Science and Technology
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    • v.14 no.1
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    • pp.85-95
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    • 2023
  • In recent years, solid-state Li metal batteries (SSLBs) have attracted significant attention as the next-generation batteries with high energy and power densities. However, uncontrolled dendrite growth and the resulting pulverization of Li during repeated plating/stripping processes must be addressed for practical applications. Herein, we report a plastic-crystal-based polymer/ceramic composite solid electrolyte (PCCE) to resolve these issues. To fabricate the one-side ceramic-incorporated PCCE (CI-PCCE) film, a mixed precursor solution comprising plastic-crystal-based polymer (succinonitrile, SN) with garnet-structured ceramic (Li7La3Zr2O12, LLZO) particles was infused into a thin cellulose membrane, which was used as a mechanical framework, and subsequently solidified by using UV-irradiation. The CI-PCCE exhibited good flexibility and a high room-temperature ionic conductivity of over 10-3 S cm-1. The Li symmetric cell assembled with CI-PCCE provided enhanced durability against Li dendrite penetration through the solid electrolyte (SE) layer than those with LLZO-free PCCEs and exhibited long-term cycling stability (over 200 h) for Li plating/stripping. The enhanced Li+ transference number and lower interfacial resistance of CI-PCCE indicate that the ceramic-polymer composite layer in contact with the Li anode enabled the uniform distribution of Li+ flux at the interface between the Li metal and CI-PCCE, thereby promoting uniform Li plating/stripping. Consequently, the Li//LiFePO4 (LFP) full cell constructed with CI-PCCE demonstrated superior rate capability (~120 mAh g-1 at 2 C) and stable cycle performance (80% after 100 cycles) than those with ceramic-free PCCE.

Effects of Donor Cell Treatments on the Production of Transgenic Cloned Piglets (공여세포 처리 조건이 형질전환 복제돼지 생산에 미치는 영향)

  • Kwon, Dae-Jin;Kwak, Tae-Uk;Oh, Keon-Bong;Kim, Dong-Hoon;Yang, Byoung-Chul;Im, Gi-Sun;Kim, Jin-Hoi;Park, Jin-Ki;Hwang, Seong-Soo
    • Reproductive and Developmental Biology
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    • v.35 no.3
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    • pp.197-201
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    • 2011
  • This study was conducted to investigate the effects of donor cell treatments on the production of transgenic cloned piglets. Ear fibroblast cell obtained from NIH MHC Inbred minipig was used as control. The GalT knock-out/CD45 knock-in (GalT/CD46) transgenic cell lines were established and used as donor cells. The reconstructed GalT/CD46 embryos were surgically transferred into oviduct of naturally cycling surrogate sows (Landrace ${\times}$ Yorkshire) on the second day of standing estrus. Unlike control (1.2 kV/cm, 75.4%), the fusion rate of the GalT/CDl6 donor cells was significantly higher in 1.5 kV/cm, (84.5%) than that of 1.25 kV/cm, (20.2%) (p<0.01). When the number of the transferred embryos were more than 129, the pregnancy and delivery rates were increased to 13/20 (65%) and 5/20 (25%) compared to less then 100 group [1/6 (16.7%) and 0/6 (0%)], respectively. To analyze the effect of donor cell culture condition on pregnancy and delivery rates, the GalT/CD46 donor cells were cultured with DMEM or serum reduced medium. In serum reduced medium group, the pregnancy and delivery rates were improved to 8/12 (66.7%) and 5/12 (41.7%) compared to DMEM group [3/7 (42.9%) and 0/7 (0%)], respectively. In conclusion, it can be postulated that an appropriate fusion condition and culture system is essential factors to increase the efficiency of the production of transgenic cloned piglets.

Effect of Trichostatin A on Anti HepG2 Liver Carcinoma Cells: Inhibition of HDAC Activity and Activation of Wnt/β-Catenin Signaling

  • Shi, Qing-Qiang;Zuo, Guo-Wei;Feng, Zi-Qiang;Zhao, Lv-Cui;Luo, Lian;You, Zhi-Mei;Li, Dang-Yang;Xia, Jing;Li, Jing;Chen, Di-Long
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.18
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    • pp.7849-7855
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    • 2014
  • Purpose: To investigate the effect of deacetylase inhibitory trichostatin A (TSA) on anti HepG2 liver carcinoma cells and explore the underlying mechanisms. Materials and Methods: HepG2 cells exposed to different concentrations of TSA for 24, 48, or 72h were examined for cell growth inhibition using CCK8, changes in cell cycle distribution with flow cytometry, cell apoptosis with annexin V-FTIC/PI double staining, and cell morphology changes under an inverted microscope. Expression of ${\beta}$-catenin, HDAC1, HDAC3, H3K9, CyclinD1 and Bax proteins was tested by Western blotting. Gene expression for ${\beta}$-catenin, HDAC1and HDAC3 was tested by q-PCR. ${\beta}$-catenin and H3K9 proteins were also tested by immunofluorescence. Activity of Renilla luciferase (pTCF/LEF-luc) was assessed using the Luciferase Reporter Assay system reagent. The activity of total HDACs was detected with a HDACs colorimetric kit. Results: Exposure to TSA caused significant dose-and time-dependent inhibition of HepG2 cell proliferation (p<0.05) and resulted in increased cell percentages in G0/G1 and G2/M phases and decrease in the S phase. The apoptotic index in the control group was $6.22{\pm}0.25%$, which increased to $7.17{\pm}0.20%$ and $18.1{\pm}0.42%$ in the treatment group. Exposure to 250 and 500nmol/L TSA also caused cell morphology changes with numerous floating cells. Expression of ${\beta}$-catenin, H3K9and Bax proteins was significantly increased, expression levels of CyclinD1, HDAC1, HDAC3 were decreased. Expression of ${\beta}$-catenin at the genetic level was significantly increased, with no significant difference in HDAC1and HDAC3 genes. In the cytoplasm, expression of ${\beta}$-catenin fluorescence protein was not obvious changed and in the nucleus, small amounts of green fluorescence were observed. H3K9 fluorescence protein were increased. Expression levels of the transcription factor TCF werealso increased in HepG2 cells following induction by TSA, whikle the activity of total HDACs was decreased. Conclusions: TSA inhibits HDAC activity, promotes histone acetylation, and activates Wnt/${\beta}$-catenin signaling to inhibit proliferation of HepG2 cell, arrest cell cycling and induce apoptosis.