• Title/Summary/Keyword: PC3 cell line

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Mutations of p53 Tumor Suppressor Gene in Human Lung Cancer Cell Lines (사람 폐암세포주에서 p53 종양억제유전자의 변이)

  • Hong, Weon-Seon;Hong, Seok-Il;Lee, Dong-Soon;Son, Young-Sook;Lee, Choon-Taek
    • Tuberculosis and Respiratory Diseases
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    • v.40 no.6
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    • pp.653-658
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    • 1993
  • Background: Recent advancement of molecular genetics has revealed that malignant transformation of a cell may be a complex multistep process and this process is grouped, in general, into two distinct categories, activation of protooncogenes and inactivation of tumor suppressor genes. This study was focused on the mutation of p53 tumor suppressor gene, because p53 gene mutation is now generally accepted to be one of the most frequent genetic changes in a variety of human cancers. Although lung cancer is one of the common cancers in Korea, the genetic change in the carcinogenesis process is not yet known clearly. To investigate the role of p53 gene mutation in lung cancer, we examined the mutations of exon 4-8 of the p53 gene in humna lung cancer cell lines, because most of the mutations of p53 gene have been reported to develop in exon 4-8. Method: Genomic DNA was obtained by the digestion of proteinase K and the extraction by phenol-chloroform-ethanol method from two human pulmonary adenocarcinoma cell lines, PC-9 and PC-14, and one human small cell lung cancer cell line, H69. To detect the mutations of exon 4-8 of the p53 gene, polymerase chain reaction single-strand conformation polymorphism(PCR-SSCP) analysis was performed with the DNA extracted from the cells. Results: The mutation of p53 gene was present in all three cell lines tested. In PC-9, PC-14 and H69, the altered mobility was detected in exon 7, 7 and 5, respectively. Conclusion: These results suggest that p53 gene mutation plays an important role in certain steps of the carcinogenesis of human non-small cell and small cell lung cancer.

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Roles of MicroRNA-21 and MicroRNA-29a in Regulating Cell Adhesion Related Genes in Bone Metastasis Secondary to Prostate Cancer

  • Mohamad, Maisarah;Wahab, Norhazlina Abdul;Yunus, Rosna;Murad, Nor AzianAbdul;Zainuddin, Zulkifli Md;Sundaram, Murali;Mokhtar, Norfilza Mohd
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.7
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    • pp.3437-3445
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    • 2016
  • Background: There is an increasing concern in the role of microRNA (miRNA) in the pathogenesis of bone metastasis (BM) secondary to prostate cancer (CaP). In this exploratory study, we hypothesized that the expression of vinculin (VCL) and chemokine X3C ligand 1 (CX3CL1) might be down-regulated in clinical samples, most likely due to the post-transcriptional modification by microRNAs. Targeted genes would be up-regulated upon transfection of the bone metastatic prostate cancer cell line, PC3, with specific microRNA inhibitors. Materials and Methods: MicroRNA software predicted that miR-21 targets VCL while miR-29a targets CX3CL1. Twenty benign prostatic hyperplasia (BPH) and 16 high grade CaP formalin-fixed paraffin embedded (FFPE) specimens were analysed. From the bone scan results, high grade CaP samples were further classified into CaP with no BM and CaP with BM. Transient transfection with respective microRNA inhibitors was done in both RWPE-1 (normal) and PC3 cell lines. QPCR was performed in all FFPE samples and transfected cell lines to measure VCL and CX3CL1 levels. Results: QPCR confirmed that VCL messenger RNA (mRNA) was significantly down-regulated while CX3CL1 was up-regulated in all FFPE specimens. Transient transfection with microRNA inhibitors in PC3 cells followed by qPCR of the targeted genes showed that VCL mRNA was significantly upregulated while CX3CL1 mRNA was significantly down-regulated compared to the RWPE-1 case. Conclusions: The down-regulation of VCL in FFPE specimens is most likely regulated by miR-21 based on the in vitro evidence but the exact mechanism of how miR-21 can regulate VCL is unclear. Up-regulated in CaP, CX3CL1 was found not regulated by miR-29a. More microRNA screening is required to understand the regulation of this chemokine in CaP with bone metastasis. Understanding miRNA-mRNA interactions may provide additional knowledge for individualized study of cancers.

Antioxidant and Anti-inflammatory Effects of Plant Extracts from Bangladesh (방글라데시 식물 추출물의 항산화 및 항염 효과 연구)

  • You, SoHyeon;Kim, Gun-Hee
    • Journal of the Korean Society of Food Culture
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    • v.35 no.6
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    • pp.605-612
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    • 2020
  • In this study, 11 plant extracts from Bangladesh were used to evaluate the total phenolic and flavonoid content, in vitro antioxidant activities using 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2-azinobis-(3-ethylbenzthiazolin-6-sulfonic acid) (ABTS), and ferric reducing antioxidant power (FRAP) assay. Also, the inhibitory effect of nitric oxide (NO) production in RAW 264.7 macrophage cell line and the neuroprotective effect on H2O2-induced PC12 cells were tested. Our results revealed that Piper betle L. showed the highest total phenolic content (162.2 mg GAE/g extract) among the 11 plants from Bangladesh. Most plants showed strong radical scavenging effects and ferric reducing antioxidant power. Besides, Piper betle L. protected PC12 neuronal cells against H2O2 related oxidative stress in LPS-induced PC12 cells. Regarding the anti-inflammatory effect, Piper betle L. significantly inhibited NO accumulation in LPS-induced RAW 264.7 cells. Our results provide evidence that Piper betle L. could be useful for the development of functional health foods.

Self Production of Radioisotope and Radiopharmaceuticals Divider (방사성동위원소 및 방사성의약품 분주장치의 자체제작)

  • Hong, Sung-Tack;Park, Kwang-Seo;Kim, Seok-Ki;Won, Woo-Jae
    • The Korean Journal of Nuclear Medicine Technology
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    • v.14 no.2
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    • pp.177-180
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    • 2010
  • Purpose: As PET test came to be covered by the pay system of medical insurance (July 1, 2006) and the needs for it becoming increased for laboratory purpose, it became necessary to purchase expensive medical equipments to solve those problems. However, as most of equipments that are operated by cyclotron are very expensive as to amount from tens of millions up to hundreds of millions of won, it is difficult to purchase those equipments from the point of medical organizations. It may be possible to self manufacture those equipments with least costs if their parts functions that meets the operators demands. The Nuclear Medicine department of National Cancer Center (NCC) is trying to manufacture and use equipments that can be made with least costs, including introducing 2 medical equipments that can improves the operator's works. Materials and Methods: Example 1: Self production of radioisotope($^{18}F$) divider was fabricated. The NCC's Nuclear Medicine department acquired one acrylic panel, seven 3-way valve, tubing etc. that can be found in the market to make the main body of divider in cooperation with biomedical engineering, and placed them inside hot cell, and installed switching box outside of hot cell to make it possible to control them from outside. This main body of divider were placed in radioisotope transfer line that are manufactured in the cyclotron. Example 2: Self production of $^{18}F$-FDG automated divider was fabricated. The NCC's Nuclear Medicine department used cavro pump syringe that consists the main body of divider in cooperation with biomedical engineering, biomedical engineering developed programs that divides a certain amount. $^{18}F$-FDG automated divider is placed inside hot cell, and cable chords were used in the equipment, and then it was connected to PC outside hot cell to make it possible to control the $^{18}F$-FDG automated divider. Results: From the NCC's Nuclear Medicine department tests that were carried out from March, 2007 until now, we found out that radioisotope can be sent to radiopharmaceuticals composite module we want, and from the tests that are carried out at NCC's Nuclear Medicine department using $^{18}F$-FDG automated divider since August, 2009 it was possible to distribute radiopharmaceuticals into vial intended. Conclusion: Through the two examples above, we found out that costs can be reduced by self manufacturing expensive equipments from NCC's cyclotron room with least costs. Also, it decreased radiation exposure dose on workers, and set up problem solving processes in cooperation with lots of parties related.

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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 Effects of Antioxidant and Anti-Alzheimer on Hydrogen peroxide and $\beta$-amyloid peptid-induced PC 12 cells by Semen Ziziphi Spinosae water extract ($H_{2}O_2$와 A$\beta$로 유도된 pc12 cell에서 생산조인(生酸棗仁) 수추출물의 항산화 및 항치매 효과)

  • Lee, Sang-Won;Kim, Dae-Hyun;Yun, Jong-Hyun;Kim, Jin-Woo;Jung, Ejun-Young;Lee, Seoung-Geun;Lee, Key-Sang;Kim, Tae-Heon;Lyu, Yeoung-Su;Kang, Hyung-Won
    • Journal of Oriental Neuropsychiatry
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    • v.19 no.3
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    • pp.179-193
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    • 2008
  • Objective: The antioxidant and anti-Alzheimer effects of Semen Ziziphi Spinosae (SZS) water extract against the amyloid beta peptide (1-42) or H202-induced oxidative damage and cell death were investigated in rat pheochromocytoma line PC 12. Methods: The cells were incubated with SZS water extract and oxidative damage-inducing materials, amyloid beta peptide (1-42) or H2O2 for 24 h. The cellular viability was assessed by WST-1 assay, cytotoxic damage by LDH activity assay, oxidative damages of cells by fluorescence spectrophotometric method, and apoptosis by TUNEL staining assay. Results and Conclusions: 1. Preincubation of the cells with SZS water extract prior to amyloid beta peptide (1-42) (2 uM) or H2O2 (30 uM) exposure elevated the cell survival close to the control and decreased the level of LDH activity and the fluorescence from the cell homogenates and TUNEL staining of the cells, compared to only amyloid beta peptide (1-42) (2 uM) or H2O2 (30 uM) treated conditions. 2. Our study suggests that Semen Ziziphi Spinosae (SZS) water extract has protective effects against amyloid beta peptide (1-42) or H2O2-induced cell toxicity through the antioxidation mechanism, which might be beneficial for the treatment of Alzheimer's disease.

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Allium monanthum Flavors, Biological Activity and Characteristics according to collecting in Different Region (달래의 향기성분, 생리활성과 채취 지역별 품질특성)

  • Oh, Tae-Seok;Kim, Chang-Ho;Cho, Yong-Koo;Kim, Sung-Min;Kim, Pom-Ho;Shin, Dong-Il
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.15 no.8
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    • pp.5176-5185
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    • 2014
  • This study was conducted from 2010 to May 2012 to determine the volatile flavor compositions, biological activity and components of A. monanthum from different regions in Korea. The flavors of A. monanthum were extracted by SPME methods and it contained forty-two compounds that included mainly hydrocarbons and acids. The cancer cell growth inhibition activities of A. monanthum on the cancer cell (HaCaT, HepG2, HCT116, PC3) line were increased in a dose-dependent manner and the hexane fraction showed the highest antiproliferation effects. A. monanthum also showed the highest antioxidant activity. The results suggest that A. monanthum can be used as bioactive and functional materials.

?Effects of Duchesnea indica on Several Kinds of Cancer Cells (사매가 수종(數種)의 암세포(癌細胞)에 미치는 영향(影響))

  • Kim, Yun-Kwan;Kim, Jin-Sung;Yoon, Sang-Hyub;Ryu, Ki-Won;Ryu, Bong-Ha
    • The Journal of Internal Korean Medicine
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    • v.26 no.2
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    • pp.320-332
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    • 2005
  • Objectives: The aim of the experiment is to identify any anti-tumor effects of Duchesnea indica(Andr.) Focke on stomach, liver, urinary bladder, prostate and kidney cancer cells. Materials & Methods: For cancer cells, AGS stomach, Hep3B and Hep3G2 liver, HT-1197, HT-1376 urinary bladder, PC3 prostate, and A-704 kidney cancer cells, all obtained from Korean Ce 11 Line Bank, were used. The boiled extract of Duchesnea indica(Andr.) Focke (10 and 20 microliters) was injected into cultures, and the cultures were observed at 0, 6 and 12 hours, and from then on at 12 hours intervals up to 72 hours. The destruction of stomach, liver, urinary bladder, prostate and kidney cancer cells were measured through Trypan blue exclusion testing. The suppresion on viability of stomach, liver, urinary bladder, prostate and kidney cancer cells was measured via MTT assay. Anti-cancer mechanisms were assessed by analyzing the cell cycle. Results: In morphologic change, AGS, Hep3B, HepG2 showed the withdrawn and floating appearance that is typical in cellular impairment. The destruction of AGS, HT-1197, HT-1376, A-704, PC-3, Hep3B and HepG2 cancer cells in each test group was greater than that in the control group to a statistically significant degree. The suppression on viability of AGS, HT-1197 and Hep3G in each test group was greater than that in the control group to a statistically significant degree. Analysis of the cell cycle after injection of D... Focke showed inhibition of cell division in all test groups(AGS, Hep3B, HepG2, HT-1197, HT-1376, PC3, A-704). Conclusions: The results of this experiment suggest that Duchesnea indica(Andr.) Focke has statistically significant anti-tumor effects on stomach, urinary bladder, kidney, prostate and liver cancer, of which stomach and liver cancer are prominently significant. This in vitro experiment supports a role for Duchesnea indica(Andr.) Focke as a potential cancer treatment, but progressive research on Duchesnea indica(Andr.) Focke and its anti-tumor effects is needed to develop a practical application for it in cancer treatment.

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Induction of Apoptosis by Treatment of Human Prostate Cancer LNCaP Cells with Methanol Fractions from Prunus mume (매실(Prunus mume) 메탄올 분획물의 처리에 따른 인체 전립선암세포 LNCaP의 apoptosis 유도 효과)

  • Kim, Hwi-gon;Kim, Jeong-Ho;Heo, Ji-An;Won, Yeong-Seon;Seo, Kwon-Il
    • Journal of Life Science
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    • v.31 no.3
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    • pp.321-329
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    • 2021
  • This study examined the growth inhibitory effect of the methanol fraction of maesil (Prunus mume) extract (MMF) on LNCaP, PC-3, and RC-58T human prostate cancer cell lines. Among these cell lines, LNCaP was the most sensitive to the inhibitory effects of MMF. Observation of the morphology and apoptotic body formation in the LNCaP cells revealed morphological changes, nuclear damage, and condensation in response to MMF treatment. The suppressive effect of MMF was related to the intrinsic apoptosis pathway, as indicated by increased expression of the pro-apoptotic proteins Bax, capase-3, capase-9, and PARP and decreased expression of the anti-apoptotic protein Bcl-2. Combined treatment with MMF and the AIF inhibitor N-phenylmalemide (N-PM) indicated that MMF treatment alone had a significant growth suppression effect. The involvement of the extrinsic apoptosis pathway was also confirmed by increased expression of AIF and Endo G. The growth suppression effect of MMF was also significant when compared to the effects of a combination of the PI3K inhibitor LY294002 and MMF. The reduced expression of PI3K, p-Akt, and p-mTOR confirmed the involvement of the PI3K/Akt/ mTOR signaling pathway in regulating the anti-proliferative properties of MMF. In conclusion, the growth suppression effect of MMF in the LNCaP human prostate cancer cell line shows the possibility of using this natural product in functional foods.

The Effect of Hwangryunhaedoktang on Proliferations of Various Human Cancer Cells (황련해독탕이 수종의 인간 암세포 증식에 미치는 영향)

  • Sung, Hyun Kyung;Min, Sang Yeon;Kim, Jang Hyun
    • The Journal of Pediatrics of Korean Medicine
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    • v.27 no.1
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    • pp.59-68
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    • 2013
  • Objectives The aim of this study is to investigate whether hwang-ryun-haedok-tang (HDT) affect proliferations of androgen-dependent LNCaP prostate cancer cells, androgen-independent PC-3, DU-145 prostate cancer cells, MCF-7 human breast cancer cells, A549, NCI-H292 human pulmonary cancer cells and K-562 human chronic myelogenous leukemia cells. Materials and Methods Effects of HDT on proliferations of each cancer cell line were investigated. 20,000 cells/well were plated in each well of 96-well culture plate. After 24 hrs, 0.01-10% of HDT in culture medium was added to cancer cells. The number of cells was counted by using SRB assay or direct cell counting method after 72 hours from drug treatment. Effect of baicalein or berebrine on proliferation was assessed according to the same method. Results (1) HDT inhibited proliferations of LNCaP, PC-3 and DU-145 prostate cancer cells. (2) HDT inhibited proliferation of MCF-7 breast cancer cells. (3) HDT also inhibited proliferations of A549, NCI-H292 pulmonary cancer cells and K-562 chronic myelogenous leukemia cells. (4) Baicalein and berberine also showed inhibitory effects on proliferations of prostate and breast cancer cells. Conclusion : HDT inhibited proliferations of human prostate, breast, pulmonary and blood cancer cells. These results suggest us the potential use of HDT as a chemopreventive or chemotherapeutic agent. Effect of HDT on human cancer should be further investigated using in vivo experimental models that can reflect pathophysiology of human cancer through another studies.