• Title/Summary/Keyword: Microarray chip

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Recovery of Genes Epigenetically Altered by the Histone Deacetylase Inhibitor Scriptaid and Demethylating Agent 5-Azacytidine in Human Leukemia Cells

  • Park, Eun-Kyung;Jeon, Eun-Hyung;Kim, In-Ho;Park, Seon-Yang
    • Genomics & Informatics
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    • v.8 no.4
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    • pp.185-193
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    • 2010
  • Histone deacetylation and demethylation are epigenetic mechanisms implicated in cancer. Studies regarding the role of modulation of gene expression utilizing the histone deacetylase inhibitor scriptaid and the demethylating agent 5-azacytidine in HL-60 leukemia cells have been limited. We studied the possibility of recovering epigenetically silenced genes by scriptaid and 5-azacytidine in human leukemia cells by DNA microarray analysis. The first group was leukemia cells that were cultured with 5-azacytidine. The second group was cultured with scriptaid. The other group was cultured with both agents. Two hundred seventy newly developed genes were expressed after the combination of 5-azacytidine and scriptaid. Twenty-nine genes were unchanged after the combination treatment of 5-azacytidine and scriptaid. Among the 270 genes, 13 genes were differed significantly from the control. HPGD, CPA3, CEACAM6, LOC653907, ETS1, RAB37, PMP22, FST, FOXC1, and CCL2 were up-regulated, and IGLL3, IGLL1, and ASS1 were down-regulated. Eleven genes associated with oncogenesis were found among the differentially expressed genes: ETS1, ASCL2, BTG2, BTG1, SLAMF6, CDKN2D, RRAS, RET, GIPC1, MAGEB, and RGL4. We report the results of our leukemia cell microarray profiles after epigenetic combination therapy with the hope that they are the starting point of selectively targeted epigenetic therapy.

Large-Circular Single-stranded Sense and Antisense DNA for Identification of Cancer-Related Genes (장환형 단일가닥 DNA를 이용한 암세포 성장 억제 유전자 발굴)

  • Bae, Yun-Ui;Moon, Ik-Jae;Seu, Young-Bae;Doh, Kyung-Oh
    • Microbiology and Biotechnology Letters
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    • v.38 no.1
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    • pp.70-76
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    • 2010
  • The single-stranded large circular (LC)-sense DNA were utilized as probes for DNA chip experiments. The microarray experiment using LC-sense DNA probes found differentially expressed genes in A549 cells as compared to WI38VA13 cells, and microarray data were well-correlated with data acquired from quantitative real-time RT-PCR. A 5K LC-sense DNA microarray was prepared, and the repeated experiments and dye swap test showed consistent expression patterns. Subsequent functional analysis using LC-antisense library of overexpressed genes identified several genes involved in A549 cell growth. These experiments demonstrated proper feature of LC-sense molecules as probe DNA for microarray and the potential utility of the combination of LC-sense microarray and antisense libraries for an effective functional validation of genes.

Construction and Analysis of a DNA Microarray for the Screening of Biosynthetic Genes of Secondary-Metabolites formation in Streptomyces (방선균 유래 이차대사 생합성 유전자 분석용 DNA Microarray 제작 및 해석)

  • Nam Soo Jung;Kang Dae-Kyung;Rhee Ki Hyeong;Kim Jong-Hee;Kang Sang Sun;Chang Yong Keun;Hong Soon-Kwang
    • Korean Journal of Microbiology
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    • v.41 no.2
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    • pp.105-111
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    • 2005
  • Streptomyces produces many kinds of secondary-metabolites including antibiotics. Screening of a new compound and elucidation of a biosynthetic pathway for the secondary metabolites are very important fields of biology, however, there is a main problem that most of the identified compounds are already researched compounds. To solve these problems, a microarray system that is based on the data related to the biosynthetic genes for secondary-metabolites was designed. For the main contents of DNA microarray, the important genes for the bio-synthesis of aminoglycosides, polyenes group, enediyne group, alpha-glucosidase inhibitors, glycopeptide group, and orthosomycin group were chosen. A DNA microarray with 69 genes that were involved in the bio-synthesis for the antibiotics mentioned above was prepared. The usability of the DNA microarray was confirmed with the chromosomal DNA and total RNA extracted from S. coelicolor whose genomic sequence had already been reported.

어류 rhabdovirus의 신속 진단을 위한 Oligonucleotide chip의 개발 2. Oligonucleotide chip을 위한 기반 조건 구축

  • 김영주;강지희;임재성;김상봉;이명숙
    • Proceedings of the Korean Society of Fisheries Technology Conference
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    • 2003.05a
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    • pp.311-312
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    • 2003
  • Microarray를 해석하는데 있어서 방해가 되는 요인에는 여러 가지가 있을 수 있다. 예를 들면 probe DNA의 농도, spotting 시의 습도, spot의 크기 및 모양, slide blocking 과정, lab리ing technique, hybri야zation 온도와 시간 그리고 background signal 등이 그것이다(Hegde et at., 2000). 본 연구에서는 전보에서 선정된 rhabdovirus의 96개 probe로 실제 oligonucleotide chip을 만들어 spot의 모양이나 크기에 관계되는 spotting 조건을 확립하고, slide blocking과 slide washing에 따른 background signal을 낮추기 위한 과정을 도입하였다. (중략)

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Oligonucleotide Chip Analysis of Cervi parvum cornu Herbal-acupuncture Solution (CPC-HAS) on SNU484 carcinomar cells (녹용약침액(鹿茸藥鍼液)의 위암세포주(胃癌細胞柱)에 대한 Oligonucleotide Chip 분석)

  • Ryu, Sung-Hyun;Lee, Kyung-Min;Lee, Bong-Hyo;Jung, Tae-Young;Seo, Jung-Chul;Lim, Seong-Chul
    • Korean Journal of Acupuncture
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    • v.23 no.2
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    • pp.125-136
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    • 2006
  • Objectives: It has long been known about the osteogenic effect of CPC-HAS(cervi parvum cornu herbal-acupuncture solution) on bone tissues. However, it has not been determined the effect of CPC-HAS on cancer cells. The purpose of this study is to screen the CPC-HAS mediated differentially expressed genes..in cancer cells such as SNU484 gastric cancer cell lines. Oligonucleotide microarray approache was employed to screen the differential expression genes. Methods: CPC-HAS was prepared by boiling and stored at $-70^{\circ}C$ until use. Cells were treated with various concentrations of CPC-HAS (0.1, 0.5, 1.5, 10, 20 mg/ml) for 24 h. Cell toxicity was tested by MTT assay. To screen the differentially expressed genes in cancer cells, cells were treated with 1.5 mg/ml of CPC-HAS. For oligonucleotide microarray assay, total RNA was used for gene expression analysis using oligonucleotide Genechip(Human genome U133 Plus 2.0., Affimatrix Co.). Results: It has no cytotoxic effects on SNU484 cell in all concentrations(0.l, 0.5, 1.5, 10, 20 mg/ml). In oligonucleotide microarray assay, in SNU484 cells, the number of more than twofold up-regulated genes was 5 while, the number of more than twofold down-regulated genes was 10. Conclusions: This study showed the screening of CPC-HAS mediated differentially regulated genes using combined approaches of oligonucleotide microarray. The screened genes will be used for the better understanding of the therapeutic effects of CPC-HAS on cancer fields.

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Learning Graphical Models for DNA Chip Data Mining

  • Zhang, Byoung-Tak
    • Proceedings of the Korean Society for Bioinformatics Conference
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    • 2000.11a
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    • pp.59-60
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    • 2000
  • The past few years have seen a dramatic increase in gene expression data on the basis of DNA microarrays or DNA chips. Going beyond a generic view on the genome, microarray data are able to distinguish between gene populations in different tissues of the same organism and in different states of cells belonging to the same tissue. This affords a cell-wide view of the metabolic and regulatory processes under different conditions, building an effective basis for new diagnoses and therapies of diseases. In this talk we present machine learning techniques for effective mining of DNA microarray data. A brief introduction to the research field of machine learning from the computer science and artificial intelligence point of view is followed by a review of recently-developed learning algorithms applied to the analysis of DNA chip gene expression data. Emphasis is put on graphical models, such as Bayesian networks, latent variable models, and generative topographic mapping. Finally, we report on our own results of applying these learning methods to two important problems: the identification of cell cycle-regulated genes and the discovery of cancer classes by gene expression monitoring. The data sets are provided by the competition CAMDA-2000, the Critical Assessment of Techniques for Microarray Data Mining.

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ALTERED GENE EXPRESSION IN RADIATION INDUCED TUMORIGENESIS OF NIH3T3 CELLS REVEALED BY MICROARRAY

  • Kang, Chang-Mo;Song, Ji-Eun;Cho, Chul-Koo;Lee, Su-Jae;Lee, Yun-Sil
    • Proceedings of the Korean Society of Toxicology Conference
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    • 2002.05a
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    • pp.81-81
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    • 2002
  • The recent development of cDNA microarray or cDNA chip technology has made it possible to analyze the expression of thousands of genes at once. In present study, we made radioresistant clones (#1 and #4) from NIH3T3 cells which are not tumorigenic and we identified 4 genes using microarray system, cdk6, cdc25B, mdm-2 and nidogene, which were altered in radiaiton resistanct NIH3T3 cells.(omitted)

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Normalization of Microarray Data: Single-labeled and Dual-labeled Arrays

  • Do, Jin Hwan;Choi, Dong-Kug
    • Molecules and Cells
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    • v.22 no.3
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    • pp.254-261
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    • 2006
  • DNA microarray is a powerful tool for high-throughput analysis of biological systems. Various computational tools have been created to facilitate the analysis of the large volume of data produced in DNA microarray experiments. Normalization is a critical step for obtaining data that are reliable and usable for subsequent analysis such as identification of differentially expressed genes and clustering. A variety of normalization methods have been proposed over the past few years, but no methods are still perfect. Various assumptions are often taken in the process of normalization. Therefore, the knowledge of underlying assumption and principle of normalization would be helpful for the correct analysis of microarray data. We present a review of normalization techniques from single-labeled platforms such as the Affymetrix GeneChip array to dual-labeled platforms like spotted array focusing on their principles and assumptions.

Genomic Detection using Electrochemical Method (전기화학적 방법에 의한 유전자의 검출)

  • Choi, Yong-Sung;Lee, Kyung-Sup;Park, Dae-Hee
    • Journal of the Korean Institute of Electrical and Electronic Material Engineers
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    • v.18 no.6
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    • pp.560-570
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    • 2005
  • In this paper, a microelectrode away DNA chip was fabricated on glass slide using photolithography technology. Several probe DNAs consisting of mercaptohexyl moiety at their 5' end were immobilized on the gold electrodes by DNA arrayer utilizing the affinity between gold and sulfu. Then target DNAs were hybridized and reacted with Hoechst 33258, which is a DNA minor groove binder and electrochemically active dye. Cyclic voltammetry in 5mA ferricyanide/ferrocyanide solution at 100 mV/s confirmed the immobilization of probe DNA on the gold electrodes. Linear sweep voltammetry or cyclic voltammetry showed a difference between target DNA and control DNA in the anodic peak current values. It was derived from Hoechst 33258 concentrated at the electrode surface through association with formed hybrid. It suggested that this DNA chip could recognize the sequence specific genes. It suggested that multichannel electrochemical DNA microarray is useful to develop a portable device for clinical gene diagnostic system.