• Title/Summary/Keyword: 2D cell lines

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Radical Scavenging Activity and Cytotoxicity of Maysin(C-glycosylflavone) isolated from Silks of Zea mays L.

  • Kim, Sun-Lim;Snook, Maurice-E;Lee, Jong-Ock
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.48 no.5
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    • pp.392-396
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    • 2003
  • Maysin, a C-glycosylflavone, was isolated from the silks of maize, Zea mays L. The ESI mass spectrum indicates that molecular weight of maysin is $577\textrm{M}^+$m/z, and the ether-linked sugar is rhamnose, $431\textrm{M}^+$m/z (MW$^{+}$-146). The DPPH (1,1-Diphenyl-2-picrylhydrazyl) radical scavenging activity of maysin was higher than that of rutin. However, as compared with its aglycon luteolin, maysin showed the relatively moderate DPPH scavenging activity mainly due to the glycosylation of two sugars moieties, keto-fucose and rhamnose. In the in vitro cytotoxicity test against the five human tumor cell lines such as lung (A549), ovarian (SK-OV-3), melanoma (SK-MEL-2), central nerve system (XF-489), and colon (HCT-15), maysin exhibited the relatively weaker activities than cisplatin. The $\textrm{ED}_{50}$ values of maysin were 62.24, 43.18, 16.83, 37.22, and 32.09/$m\ell$, respectively. Result suggests that maysin is a potential cytotoxicity compound, particularly for human colon, central nerve system, and melanoma tumors.s.

A Cytotoxic Secocycloartenoid from Abies koreana

  • Kim, Hyun-Jung;Le, Quoc-Khanh;Lee, Mi-Hyun;Kim, Tae-Sung;Lee, Hyeong-Kyu;Kim, Young-Ho;Bae, Ki-Hwan;Lee, Ik-Soo
    • Archives of Pharmacal Research
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    • v.24 no.6
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    • pp.527-531
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    • 2001
  • Two triterpenoids, 24-methylene-3,4-seco-cycloart-4(28)-en-3-oic acid (1) and 3-oxo-$9{\beta}$-lanosta-7,22Z,24-trien-26,23-olive (6) were isolated from Abies koreana, together with $\beta$-sitosterol (2), maltol (3), ${\beta}-sitosterol-O-{$\beta}-D-glucoside$ (4), and hexacosylferulate (5). The structures of the compounds were established based on the spectroscopic data. The cytotoxic activities of triterpenoids have been evaluated using the sulforhodamine B (SRB) method. Compound 1 showed moderate cytotoxicities against human lung carcinoma (A549), ovarian carcinoma (SK-OV-3), malignant melanoma (SK-MEL-2), and colon carcinoma (HCT-15) cell lines.

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Cytotoxic Isoflavanones from Uraria clarkei

  • Huang, Xiang-Zhong;Bai, Xi-Shan;Liang, Hui;Wang, Chao;Li, Wen-Juan;Guo, Jun-Ming;Jiang, Zhi-Yong
    • Bulletin of the Korean Chemical Society
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    • v.34 no.5
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    • pp.1421-1424
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    • 2013
  • Two new isoflavanones, (3R) 5,7,3',4'-tetrahydroxy-2'-methoxyisoflavanone (1) and (3R) 5',8-di-(${\gamma}$,${\gamma}$-dimethylallyl)-2',5-dihydroxyl-4',7-dimethoxyl-isoflavanone (2), were isolated from Uraria clarkei, together with two known compounds dalbergioidin (3), 5,7-dihydroxy-2',4'-dimethoxyisoflavanone (4). The structures involving the absolute configuration of the new compounds were well elucidated by MS, IR, UV, CD, 1D and 2D NMR analyses. Cytotoxicity of the four compounds were assessed, results suggested that compound 2 possessed well cytotoxic activity, against the Hela, K562, and HL60 cell lines with $IC_{50}$ values of 28.0, 40.6 and $35.1{\mu}M$, respectively.

The transposition pattern of the Ac element and its use for targeted transposition in Arabidopsis thaliana

  • Machida, Yasunori;Onouchi, Hitoshi;Tanaka, Hirokazu;Hamada, Susumu;Ishikawa, Takaaki;Semiarti, Endang;Iwakawa, Hidekazu;Nomura, Kiyohito;Machida, Chiyoko
    • Proceedings of the Botanical Society of Korea Conference
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    • 1999.07a
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    • pp.11-15
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    • 1999
  • In order to evlauate feasibility of the gene tagging by the maize transposable element Ac in heterologous plant systems, we have investigated physical distances and directions of transposition of the element in Arabidopsis thaliana and tobacco cultured cell line BY-2. We prepared a T-DNA construct that carried a non-autonomous derivative of Ac with a site for cleavage by endonuclease I-Scel (designated dAc-I-RS element). Another cleavage site was also introduced into the T-DNA region outside dAc-I-RS. A number of transgenic Arabidopsis plants were generated, each of which had a single copy of the T-DNA at a different chromosomal location. To examine the pattern of transposition, three out of these transgenic plants were crossed with the Arabidopsis plant that carried the gene for Ac transposase and progeny in which dAc-I-RS had been transposed were isolated. After digestion of the genomic DNA of these progeny with I-SceI, sizes of segment of DNA were determined byd pulse-field gel electrophoresis. We also performed linkage analysis for the transposed elements and sites of mutations near the elements. Our results with three transgenic lines showed that 50% of all transposition events had occurred within 1,700 kilo-base pairs (kb) on the same chromosome, with 35% within 200 kb, and that the elements transposed in both directions on the chromosome with roughly equal probability. The data thus indicate that the Ac-Ds system is most useful for tagging of genes that are present within 200 kb of the chromosomal site of Ac in Arabidopsis. In addition, determination of the precise localization of the transposed dAc-I-RS element should definitely assist in map-based cloning of genes around insertion sites. In the present paper, we report typical examples of such gene isolation studies.

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세포내의 물의 상태

  • 강사욱
    • Proceedings of the Botanical Society of Korea Conference
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    • 1985.08b
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    • pp.51-57
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    • 1985
  • In order to evlauate feasibility of the gene tagging by the maize transposable element Ac in heterologous plant systems, we have investigated physical distances and directions of transposition of the element in Arabidopsis thaliana and tobacco cultured cell line BY-2. We prepared a T-DNA construct that carried a non-autonomous derivative of Ac with a site for cleavage by endonuclease I-Scel (designated dAc-I-RS element). Another cleavage site was also introduced into the T-DNA region outside dAc-I-RS. A number of transgenic Arabidopsis plants were generated, each of which had a single copy of the T-DNA at a different chromosomal location. To examine the pattern of transposition, three out of these transgenic plants were crossed with the Arabidopsis plant that carried the gene for Ac transposase and progeny in which dAc-I-RS had been transposed were isolated. After digestion of the genomic DNA of these progeny with I-SceI, sizes of segment of DNA were determined byd pulse-field gel electrophoresis. We also performed linkage analysis for the transposed elements and sites of mutations near the elements. Our results with three transgenic lines showed that 50% of all transposition events had occurred within 1, 700 kilo-base pairs (kb) on the same chromosome, with 35% within 200 kb, and that the elements transposed in both directions on the chromosome with roughly equal probability. The data thus indicate that the Ac-Ds system is most useful for tagging of genes that are present within 200 kb of the chromosomal site of Ac in Arabidopsis. In addition, determination of the precise localization of the transposed dAc-I-RS element should definitely assist in map-based cloning of genes around insertion sites. In the present paper, we report typical examples of such gene isolation studies.

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Two New Diphenylethylenes from Arundina graminifolia and Their Cytotoxicity

  • Li, Yin-Ke;Zhou, Bin;Ye, Yan-Qing;Du, Gang;Niu, De-Yun;Meng, Chun-Yang;Gao, Xue-Mei;Hu, Qiu-Fen
    • Bulletin of the Korean Chemical Society
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    • v.34 no.11
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    • pp.3257-3260
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    • 2013
  • Two new diphenylethylenes, gramniphenols H and I (1 and 2), together with six known diphenylethylenes (3-8), were isolated from Arundina graminifolia. The structures of 1-8 were elucidated by spectroscopic methods including extensive 1D- and 2D-NMR techniques. Compounds 1 and 2 were evaluated for their cytotoxicity against five human tumor cell lines. Compound 1 showed cytotoxicity against PC3 cells with $IC_{50}$ value of 3.5 ${\mu}M$. Compound 2 showed cytotoxicity against NB4 and PC3 cells with $IC_{50}$ values of 3.6 and 3.8 ${\mu}M$, respectively.

Real-Time PCR for Validation of Minute Virus of Mice Safety during the Manufacture of Mammalian Cell Culture-Derived Biopharmaceuticals (세포배양 유래 생물의약품 생산 공정에서 Minute Virus of Mice 안전성 검증을 위한 Real-Time PCR)

  • Lee, Dong-Hyuck;Cho, Hang-Mee;Kim, Hyun-Mi;Lee, Jung-Suk;Kim, In-Seop
    • Microbiology and Biotechnology Letters
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    • v.36 no.1
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    • pp.12-20
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    • 2008
  • Validation of viral safety is essential in ensuring the safety of mammalian cell culture-derived biopharmaceuticals, because numerous adventitious viruses have been contaminated during the manufacture of the products. Mammalian cells are highly susceptible to minute virus of mice(MVM), and there are several reports of MVM contamination during the manufacture of biopharmaceuticals. In order to establish the validation system for the MVM safety, a real-time PCR method was developed for quantitative detection of MVM in cell lines, raw materials, manufacturing processes, and final products as well as MVM clearance validation. Specific primers for amplification of MVM DNA was selected, and MVM DNA was quantified by use of SYBR Green I. The sensitivity of the assay was calculated to be $6{\times}10^{-2}TCID_{50}/mL$. The real-time PCR method was proven to be reproducible and very specific to MVM. The established real-time PCR assay was successfully applied to the validation of Chinese hamster ovary (CHO) cell artificially infected with MVM. MVM DNA could be Quantified in CHO cell as well as culture supernatant. When the real-time PCR assay was applied to the validation of virus removal during a virus filtration process, the result was similar to that of virus infectivity assay. Therefore, it was concluded that this rapid, specific, sensitive, and robust assay could replace infectivity assay for detection and clearance validation of MVM.

Apoptosis Induced by Adenosine 5'-triphosphate in Mouse Leukemic Cells (Mouse Leukemia 세포에서 Adenosine 5'-triphosphate에 의한 Apoptosis)

  • Joo, Nan-Young;Park, Kyu-Sang;Chung, Hae-Sook;Kong, In-Deok;Lee, Joong-Woo
    • The Korean Journal of Physiology and Pharmacology
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    • v.1 no.6
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    • pp.817-824
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    • 1997
  • Extracellular ATP elicits various biological responses and plays a significant role in physiological regulation. Recently, ATP-induced growth inhibitions were reported in some tumor cell lines, but these effects and mechanisms are not well hewn. This study was conducted to investigate ATP-induced growth inhibition in mouse $leukemic(P388D_1)$ cells. ATP inhibited cell growth in a dose-dependent manner as analyzed by MTS assay$(IC_{50}: 33.1\;{\mu}M)$. Nucleotides other than ATP, such as ADP$(37.5;{\mu}M)$ and AMP$(33.2;{\mu}M)$ had the same effects as ATP but adenosine$(57.8;{\mu}M)$ showed less effect than ATP. ATP attenuated the cells in $G_0/G_l\;and\;G_2/M$ phases but increased those in S phase in flow cytometric analysis. Hypodiploid cells$(A_0)$, the presumptive findings of apoptosis, were found among the ATP-treated cells. ATP induced DNA fragmentation into $180{\mu}200\;bps $as measured by electrophoresis. some apoptotic cells were stained by TUNEL method. ATP increased the intracellular free $Ca^{++}$concentration$([Ca^{++}]_i)$ and the increment of $([Ca^{++}]_i)$ was caused by influx from the extracellular space. These results suggest that extracellular ATP induces growth inhibition through apoptosis.

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Identifying Differentially Expressed Genes and Screening Small Molecule Drugs for Lapatinib-resistance of Breast Cancer by a Bioinformatics Strategy

  • Zhuo, Wen-Lei;Zhang, Liang;Xie, Qi-Chao;Zhu, Bo;Chen, Zheng-Tang
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.24
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    • pp.10847-10853
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    • 2015
  • Background: Lapatinib, a dual tyrosine kinase inhibitor that interrupts the epidermal growth factor receptor (EGFR) and HER2/neu pathways, has been indicated to have significant efficacy in treating HER2-positive breast cancer. However, acquired drug resistance has become a very serious clinical problem that hampers the use of this agent. In this study, we aimed to screen small molecule drugs that might reverse lapatinib-resistance of breast cancer by exploring differentially expressed genes (DEGs) via a bioinformatics method. Materials and Methods: We downloaded the gene expression profile of BT474-J4 (acquired lapatinib-resistant) and BT474 (lapatinib-sensitive) cell lines from the Gene Expression Omnibus (GEO) database and selected differentially expressed genes (DEGs) using dChip software. Then, gene ontology and pathway enrichment analyses were performed with the DAVID database. Finally, a connectivity map was utilized for predicting potential chemicals that reverse lapatinib-resistance. Results: A total of 1, 657 DEGs were obtained. These DEGs were enriched in 10 pathways, including cell cycling, regulation of actin cytoskeleton and focal adhesion associate examples. In addition, several small molecules were screened as the potential therapeutic agents capable of overcoming lapatinib-resistance. Conclusions: The results of our analysis provided a novel strategy for investigating the mechanism of lapatinib-resistance and identifying potential small molecule drugs for breast cancer treatment.

Novel SIRT Inhibitor, MHY2256, Induces Cell Cycle Arrest, Apoptosis, and Autophagic Cell Death in HCT116 Human Colorectal Cancer Cells

  • Kim, Min Jeong;Kang, Young Jung;Sung, Bokyung;Jang, Jung Yoon;Ahn, Yu Ra;Oh, Hye Jin;Choi, Heejeong;Choi, Inkyu;Im, Eunok;Moon, Hyung Ryong;Chung, Hae Young;Kim, Nam Deuk
    • Biomolecules & Therapeutics
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    • v.28 no.6
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    • pp.561-568
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    • 2020
  • We examined the anticancer effects of a novel sirtuin inhibitor, MHY2256, on HCT116 human colorectal cancer cells to investigate its underlying molecular mechanisms. MHY2256 significantly suppressed the activity of sirtuin 1 and expression levels of sirtuin 1/2 and stimulated acetylation of forkhead box O1, which is a target protein of sirtuin 1. Treatment with MHY2256 inhibited the growth of the HCT116 (TP53 wild-type), HT-29 (TP53 mutant), and DLD-1 (TP53 mutant) human colorectal cancer cell lines. In addition, MHY2256 induced G0/G1 phase arrest of the cell cycle progression, which was accompanied by the reduction of cyclin D1 and cyclin E and the decrease of cyclin-dependent kinase 2, cyclin-dependent kinase 4, cyclin-dependent kinase 6, phosphorylated retinoblastoma protein, and E2F transcription factor 1. Apoptosis induction was shown by DNA fragmentation and increase in late apoptosis, which were detected using flow cytometric analysis. MHY2256 downregulated expression levels of procaspase-8, -9, and -3 and led to subsequent poly(ADP-ribose) polymerase cleavage. MHY2256-induced apoptosis was involved in the activation of caspase-8, -9, and -3 and was prevented by pretreatment with Z-VAD-FMK, a pan-caspase inhibitor. Furthermore, the autophagic effects of MHY2256 were observed as cytoplasmic vacuolation, green fluorescent protein-light-chain 3 punctate dots, accumulation of acidic vesicular organelles, and upregulated expression level of light-chain 3-II. Taken together, these results suggest that MHY2256 could be a potential novel sirtuin inhibitor for the chemoprevention or treatment of colorectal cancer or both.