• Title/Summary/Keyword: PC3 cells

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Global Histidine Phosphoproteomics in Human Prostate Cancer Cells

  • Gao, Yan;Kim, Doeun;Sung, Eunji;Tan, Minjia;Kwon, Tae Gyun;Lee, Jun Nyung;Lee, Sangkyu
    • Mass Spectrometry Letters
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    • v.11 no.3
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    • pp.52-58
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    • 2020
  • Histidine phosphorylation (pHis) is increasingly recognized as an important post translational modification (PTM) in regulating cellular functions in eukaryotes. In order to clarify the role of pHis in mammalian cell signaling system, a global phosphorylation study was performed in human prostate cancer cells, PC-3M, using a TiO2 affinity chromatography. A total number of 307 pHis sites were identified on the 268 proteins among total identified 9,924 phosphorylation sites on 3,316 proteins. In addition, 22 pHis proteins were classified in enzyme category. This report provides the first database for the study of pHis in prostate cancer cells.

Bcl-2 Overexpression Inhibits Generation of Intracellular Reactive Oxygen Species and Blocks Adriamycin-induced Apoptosis in Bladder Cancer Cells

  • Kong, Chui-Ze;Zhang, Zhe
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.2
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    • pp.895-901
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    • 2013
  • Resistance to induction of apoptosis is a major obstacle for bladder cancer treatment. Bcl-2 is thought to be involved in anti-apoptotic signaling. In this study, we investigated the effect of Bcl-2 overexpression on apoptotic resistance and intracellular reactive oxygen species (ROS) generation in bladder cancer cells. A stable Bcl-2 overexpression cell line, BIU87-Bcl-2, was constructed from human bladder cancer cell line BIU87 by transfecting recombinant Bcl-2 [pcDNA3.1(+)-Bcl-2]. The sensitivity of transfected cells to adriamycin (ADR) was assessed by MTT assay. Apoptosis was examined by flow cytometry and acridine orange fluorescence staining. Intracellular ROS was determined using flow cytometry, and the activities of superoxide dismutase (SOD) and catalase (CAT) were also investigated by the xanthinoxidase and visible radiation methods using SOD and CAT detection kits. The susceptibility of BIU87-Bcl-2 cells to ADR treatment was significantly decreased as compared with control BIU87 cells. Enhanced expression of Bcl-2 inhibited intracellular ROS generation following ADR treatment. Moreover, the suppression of SOD and CAT activity induced by ADR treatment was blocked in the BIU87-Bcl-2 case but not in their parental cells. The overexpression of Bcl-2 renders human bladder cancer cells resistant to ADR-induced apoptosis and ROS might act as an important secondary messenger in this process.

The role of the voltage-gated potassium channel, Kv2.1 in prostate cancer cell migration

  • Park, Hyun Woo;Song, Min Seok;Sim, Hun Ju;Ryu, Pan Dong;Lee, So Yeong
    • BMB Reports
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    • v.54 no.2
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    • pp.130-135
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    • 2021
  • Voltage-gated potassium (Kv) channels are involved in many important cellular functions and play pivotal roles in cancer progression. The expression level of Kv2.1 was observed to be higher in the highly metastatic prostate cancer cells (PC-3), specifically in their membrane, than in immortalized prostate cells (WPMY-1 cells) and comparatively less metastatic prostate cancer cells (LNCaP and DU145 cells). However, Kv2.1 expression was significantly decreased when the cells were treated with antioxidants, such as N-acetylcysteine or ascorbic acid, implying that the highly expressed Kv2.1 could detect reactive oxygen species (ROS) in malignant prostate cancer cells. In addition, the blockade of Kv2.1 with stromatoxin-1 or siRNA targeting Kv2.1 significantly inhibited the migration of malignant prostate cancer cells. Our results suggested that Kv2.1 plays an important role as a ROS sensor and that it is a promising therapeutic molecular target in metastasis of prostate cancer.

Effects of Opuntia ficus-indica var. saboten Ripe Fruits on Protection of Neuronal PC-12 Cells and Cholinesterase Inhibition (백년초의 PC-12 신경세포 보호 및 콜린가수분해효소(cholinesterase) 저해 효과)

  • Hwang, Jeong-Seung;Im, Sungbin;Lee, Inil;Kim, Tae-Rahk;Kim, Dae-Ok
    • Korean Journal of Food Science and Technology
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    • v.48 no.1
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    • pp.86-91
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    • 2016
  • Oxidative stress caused by reactive oxygen species is ascribed to many neurodegenerative diseases like Alzheimer's disease. Phenolic antioxidants can reduce the oxidative stress. In this study, ripe fruits of Opuntia ficus-indica var. saboten (OFS) were extracted using 80% (v/v) aqueous ethanol. Total phenolic and flavonoid contents of the OFS fruits (100 g) were 409.9 mg gallic acid equivalents and 72.2 mg catechin equivalents, respectively. The OFS fruits had antioxidant capacity at 381.2, 298.2, and 3,219.9 mg vitamin C equivalents/100 g in ABTS, DPPH, and ORAC assays, respectively. The OFS fruits showed protective effects on PC-12 cells against oxidative stress in a dose-dependent manner, partly due to decrease of intracellular oxidative stress. Furthermore, the OFS fruits inhibited both acetylcholinesterase and butyrylcholinesterase. Consequently, these results suggest that the OFS fruits might be served as a source of functional materials to reduce oxidative stress in neuronal cells and to inhibit cholinesterases.

A Cyclin-Dependent Kinase Inhibitor, p16^{INK4A}, Induces Apoptosis in The Human Cancer Cells. (Cyclin-dependent Kinase저해 단백질 p16^{INK4A}의 인체 암세포에서의 세포사멸 유도 활성)

  • 김민경;이철훈
    • Microbiology and Biotechnology Letters
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    • v.32 no.1
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    • pp.72-77
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    • 2004
  • Previously, we synthesized a novel Cyclin-dependent kinase inhibitor, MCS-5A. Also, we investigated the involvement of cell cycle regulatory events during MCS-5A-mediated apoptosis in HL-60(+p16/-p53) cells with up-regulation of p16 protein expression. In contrast, apoptosis was not observed in A549(-p16/+p53) cells. Therefore we propose that $p16^{INK4A}$ is a key enzyme for inducing apoptosis. In the present studies, we have explored the mechanism of $p16^{INK4A}$ -mediated cytotoxicity and the role of p16.sup INK4A/ overexpression in the induction of apoptosis in human tumor cells. The tumor suppressor gene $p16^{INK4A}$ is known as a cyclin-dependent kinase inhibitor (CKI) and cell cycle regulator. We expressed wild type $p16^{INK4A}$ in pcDNA3.1 vector and then transfected into non-small cell lung cancer (NSCLC) cell expressing different statue of p16$^{INK4A}$, p53 gene〔A549(-p16/+p53), H1299(-p16/-p53) and HeLa(+pl6/+p53) cell line〕. TUNEL assay (including propidium iodide staining following transfection of these cell line with pcDNA3.1-pl6) indicate that p16$^{INK4A}$-mediated cytotoxicity was associated with apoptosis. This is supported by studies demonstrating an induction of caspase 3 cleavage due to the transfection of A549, H1299 and HeLa cells with pcDNA3.1-pl6. These results suggest that p16$^{INK4A}$ has a new function of inducing apoptosis which is not related with the function of tumor suppressor gene p53.

Deodorizing Effect of Several Antibacterial Medicinal Herbs on Oral Malodor (항균작용을 가진 수종 한약재의 구취억제 효과)

  • Kim, Hyun-Kyung;Park, Jae-Woo;Yoon, Seong-Woo;Ryu, Bong-Ha;Kim, Jin-Sung
    • The Journal of Korean Medicine
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    • v.31 no.4
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    • pp.151-163
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    • 2010
  • Objective: We investigated the oral malodor inhibitory effect of Scutellariae Radix (SR), Phellodendri Cortex (PC), Moutan Cortex (MTC) and Magnoliae Cortex (MGC) for the development of a gargle solution. Methods: 1. Against P. gingivalis and Pr. intermedia, the minimal bactericidal concentration (MBC) and the change of viable cells that were exposed to 1% each herbal extract were observed. 2. Deodorizing activity of 2% herbal extract and Garglin $Mint^{(R)}$ against methyl mercaptan were evaluated by gas chromatography (GC). 3. We used the salivary sediment system (SSS) as the malodor model. 4. The clinical examination was repeated 3 times by 2 subjects by $Halimeter^{(R)}$. Baseline VSC of each subject was measured. Then, the control subject gargled with cysteine for 30 sec. After 4 min, subjects would gargle for 30 seconds with herbal extracts (2%) and Garglin $Mint^{(R)}$. Subsequently, the concentration of VSC was measured at 0, 4, 8, 12, 16, 20, 40 and 60 minutes. Results: 1. Against P. gingivalis, MBC of SR, PC and MTC was 0.1%, and MBC of MGC was 1%. Removal time of P. gingivalis was as follows; 5 hr in MGC, 24 hr in SR and PC, and 48 hr in MTC. Against Pr. intermedia, MBC of SR and PC was 0.5%, and MBC of MTC, MGC was 1%. Removal time of Pr. intermedia was as follows; 5 hr in MTC and 24 hr in SR, PC and MGC. 2. Deodorizing effect of herbal extracts against methyl mercaptan was as follows; MGC and MTC had 100%, SR had 82.22%, PC had 66.60%, Garglin $Mint^{(R)}$ had 40.54%. 3. In the experiment using SSS, PC and MTC had statistically significant malodor-inhibitory effects (p<.05). 4. In the clinical examination, PC and MGC had statistically significant inhibitory effects at every elapsed time compared to the control subject. MTC had that until 40 min. SR had that at 0, 4, 8, 20, and 60 min. Conclusions: SR, PC, MTC and MGC have an antibacterial effect and the chemical removable activity of the oral malodor caused by VSC. These four herbs could have potential as effective anti-malodor agents.

Characteristics of Lithium Metal Secondary Battery Using PAN Gel-electrolyte Mixed with TiO2 Ceramic Filler (TiO2 Ceramic Filler가 혼합된 젤상의 PAN 고분자 전해질을 이용한 리튬금속 이차전지의 특성)

  • Lim, Hyo-Sung;Kim, Hyung-Sun;Cho, Byung-Won;Lee, Tae-Hee
    • Journal of the Korean Electrochemical Society
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    • v.5 no.3
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    • pp.106-110
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    • 2002
  • Gel-type polyacrylonitrile(PAN) polymer electrolytes have been prepared using ethylene carbonate(EC), propylene carbonate(PC) and dimethyl carbonate(DMC) plasticizer, $LiPF_6$ salt and $TiO_2$ ceramic filler. Electrochemical properties, such as electrochemical stability, ionic conductivity and compatibility with lithium metal and mechanical properly of polymer electrolytes were investigated. Charge/discharge performance of lithium secondary battery using these polymer electrolytes were investigated. The maximum load that the polymer electrolyte resists increased about two times as a result of adding $TiO_2$ in the polymer electrolyte containing EC and PC. Polymer electrolyte containing EC, PC and $TiO_2$ also showed ionic conductivity of $2\times10^{-3} S/cm$ at room temperature and electrochemical stability window up to 와 4.5V. Polymer electrolyte containing EC, PC, and $TiO_2$ showed the most stable interfacial resistance of $130\Omega$ during 20 days in the impedance spectra of the cells which were constructed by lithium metals as electrodes. Lithium metal secondary battery which employed $LiCoO_2$ cathode, lithium metal anode and $TiO_2$-dispersed polymer electrolyte showed $90\%$ of charge/discharge efficiency at the 1C rate of discharge.

BRI3 associates with SCG10 and attenuates NGF-induced neurite outgrowth in PC12 cells

  • Gong, Yanhua;Wu, Jing;Qiang, Hua;Liu, Ben;Chi, Zhikai;Chen, Tao;Yin, Bin;Peng, Xiaozhong;Yuan, Jiangang
    • BMB Reports
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    • v.41 no.4
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    • pp.287-293
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    • 2008
  • In a yeast two-hybrid screen, we identified the microtubule-destabilizing protein SCG10 as a potential effector protein of $BRI_3$. The association was verified using GST pull-down, Co-IP, and their perinuclear co-localization. The analysis of in vitro microtubule polymerization/depolymerization showed that the binding of $BRI_3$ to SCG10 effectively blocked the ability of SCG10 to induce microtubule disassembly, as determined by turbidimetric assays. In intact PC12 cells, $BRI_3$ exhibited the ability to stabilize the microtubule network and attenuate the microtubule-destabilizing activity of SCG10. Furthermore, co-expression of $BRI_3$ with SCG10 attenuated SCG10-mediated PC12 cell neurite outgrowth induced by NGF. These results identify a novel connection between a neuron-specific BRI protein and the cytoskeletal network, suggesting possible roles of BRI3 in the process of neuronal differentiation.

Effect of Mycelial Extract of Clavicorona pyxidata on Acetylcholinesterase and ${\beta}$-Secretase Activity in vitro

  • Lee, Tae-Hee;Park, Young-Il;Han, Yeong-Hwan
    • Journal of Microbiology
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    • v.44 no.5
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    • pp.502-507
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    • 2006
  • In a previous study, an extract of Clavicorona pyxidata DGUM 29005 mycelia demonstrated an inhibitory effect against enzyme-associated perceptual disorders. We have attempted to determine whether this mycelial extract is also capable of inhibiting the activities of acetylcholinesterase (AChE) and ${\beta}$-secretase (BACE) activity. Butanol, ethanol, and water extracts of C. pyxidata DGUM 29005 mycelia were shown to inhibit AChE activity by 99.3%, 93.7%, and 91.7%, respectively. The inhibitory value of the butanol extract was more profound than that of tacrine (95.4%). The ethanol extract also exerted an inhibitory effect against BACE activity; this fraction may harbor the potential for development into a pharmocotherapeutic modality for the treatment of Alzheimer's disease (AD) patients. Rat pheochromocytoma PC12 cells in culture were not determined to be susceptible to the cytotoxic activity evidenced by the mycelial extract. The ethanol extract inhibited endogenous AChE activity in PC12 cellular homogenates, with an $IC_{50}\;of\;67.5{\mu}g/ml$, after incubation with intact cells, and also inhibited BACE activity in a dose-dependent fashion. These results suggest that the C. pyxidata mycelial extract has the potential to enhance cholinergic function and, therefore, may perform a function in the amelioration of the cholinergic deficit observed in cases of AD, as well as other types of age-associated memory impairment.

Enantio-Selective Inhibition of (1R,9S)- and (1S,9R)-$\beta$-Hydrastines on Dopamine Biosynthesis in PC12 Cells

  • Yin, Shou-Yu;Kim, Yu-Mi;Lee, Jae-Joon;Jin, Chun-Mei;Yang, You-Jong;Kang, Min-Hee;Lee, Myung-Koo
    • Proceedings of the PSK Conference
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    • 2003.04a
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    • pp.138.2-139
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    • 2003
  • The inhibitory effects of (1R, 9S)- and (1S, 9R)-enantiomers of $\beta$-hydrastine (BHS) on dopamine biosynthesis in PC12 cells were investigated. (1R, 9S)-BHS decreased the intracellular dopamine content with the $IC_{50}$ value of 14.3 $\mu\textrm{M}$ at 24 h, but (1S, 9R)-BHS did not. In these conditions, (1R, 9S)-BHS inhibited TH activity mainly in a concentration-dependent manner(33% inhibition at 20 $\mu\textrm{M}$) and decreased TH mRNA level. (omitted)

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