• Title/Summary/Keyword: GeneChip analysis

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Molecular Biomarkers of Octachlorostyrene Exposure in Medaka, Oryzias latipes, using Microarray Technique (Microarray를 이용한 Octachlorostyrene-노출 송사리(Oryzias latipes)에서의 분자생물학적 지표연구)

  • You Dae-Eun;Kang Misun;Park Eun-Jung;Kim IL-Chan;Lee Jae-Seong;Park Kwangsik
    • Environmental Analysis Health and Toxicology
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    • v.20 no.2 s.49
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    • pp.187-194
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    • 2005
  • Octachlorostyrene (OCS) is a primarily concerning chemical in many countries because of its persistent and bioaccumulative properties in the environment. OCS is not commercially manufactured or used but it may be produced during incineration or chemical synthetic processes involving chlorinated compounds. There are several reports that OCS was found in the waters, sediments, fish, mussels, and also in human tissues. However, systematic studies on the OCS toxicities are scarce in literature. In this study, we tried to get the gene expression data using medaka DNA chip to identify biomarkers of OCS exposure. Medaka (Oryzias latipes.) was exposed to OCS 1 ppm for 2 days and 10 days, respectively. Total RNA was extracted and purified by guanidine thiocyanate method and the Cy3- and Cy5-labelled cDNAs produced by reverse trancription of the RNA were hybridized to medaka microarray. As results, eighty five genes were found to be down-or up regulated by OCS. Some of the genes were listed and confirmed by real-time PCR.

DNA Chip을 이용한 Transcriptional Activation Mechanism 분석

  • 김영준
    • Proceedings of the Korean Society for Bioinformatics Conference
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    • 2001.10a
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    • pp.45-60
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    • 2001
  • . Mediator of transcriptional regulation is the evolutionary conserved coactivator complex that plays He central role in the integration and recruitment of diverse regulatory signals and transcription machinery to certain promoters. In yeast, each Mediator subunit is required for transcriptional regulation of a distinct group of genes. In order to decipher the mechanistic roles of Mediator proteins in regulating developmental specific gene expression, we isolated, and analyzed a multiprotein complex containing Drosophila Mediate. homologs (dMediato.). dMediato. interacts with several sequence-sperific transcription factors and basal transcription machinery, and is critical for activated transcription in response to diverse transcriptional activators. In order to elucidate the function of Mediator in metazoan development, we isolated mutants of a conserved Mediate. subunit, Drosophila Med6 (dMed6). dMed6 null homozygotes failed to pupate and died in the third larval instar. Larval mitotic cells and most imaginal discs showed severe defects in proliferation, but no apparent morphological defect was observed in other larval tissues. Clonal analysis of dMed6 mutant cells revealed that dMed6 is essential for cell viability and proliferation of most adult cell types. Drosophila cDNA microarray, quantitative RT-PCR, and in situ expression analyses of developmentally regulated genes in dMed6 mutants showed that transcriptional activation of a subset of genes involved in neuroblast proliferation in the larval brain were most affected. Our results suggest that dMed6 is required in most for transcriptional regulation of a subset of genes important for cell proliferation and metabolism.

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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.

Effects of Lonicerae Caulis (LC) on Gene Expression of Human melanoma cells (인동등(忍冬藤)이 인간 유래 악성 흑색종 세포의 유전자 발현에 미치는 영향)

  • Kim, Dae-Su;Choi, Jeong-Hwa;Kim, Jong-Han;Park, Soo-Yeon;kang, Seong-In
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.22 no.1
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    • pp.11-32
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    • 2009
  • Objective : This study was designed to investigate anti-cancer and whitening activities (LC). So it was investigated the effects of LC on proliferation rates of melanoma genetic profile by LC. Methods : The genetic profile for the effect of LC on human derived melanoma cell, SK-MEL-2, was measured using microarray technique, and the functional analysis on these genes were conducted. Total 441 genes were up-regulated and 830 genes down-regulated in cells treated with LC. Genes induced or suppressed by LC were all mainly concerned with basic signalling pathways, which are involved in cell growth, differentiation and migration. Especially, many genes, which are related in apoptosis and cell cycle arrest were up-regulated by treatment with LC, and genes related in cell cycle were down-regulated. Result : The network of total protein interactions were identified by using cytoscape program, and some key molecules, such as BCL2L1, SIN3A, SMAD2 and c-myc that can be used for elucidation of therapeutical mechanism of medicine in the future. Conclusion : These results suggest possibility of LC as addition drug and whitening cosmetics. In addition, it was also suggested that related mechanisms are involved in BCL2L1, SIN3A, SMAD2 and c-myc related signalling pathways.

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Detection and Analysis of DNA Hybridization Characteristics by using Thermodynamic Method (열역학법을 이용한 DNA hybridization 특성 검출 및 해석)

  • Kim, Do-Gyun;Gwon, Yeong-Su
    • The Transactions of the Korean Institute of Electrical Engineers C
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    • v.51 no.6
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    • pp.265-270
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    • 2002
  • The determination of DNA hybridization reaction can apply the molecular biology research, clinic diagnostics, bioengineering, environment monitoring, food science and application area. So, the improvement of DNA hybridization detection method is very important for the determination of this hybridization reaction. Several molecular biological techniques require accurate predictions of matched versus mismatched hybridization thermodynamics, such as PCR, sequencing by hybridization, gene diagnostics and antisense oligonucleotide probes. In addition, recent developments of oligonucleotide chip arrays as means for biochemical assays and DNA sequencing requires accurate knowledge of hybridization thermodynamics and population ratios at matched and mismatched target sites. In this study, we report the characteristics of the probe and matched, mismatched target oligonucleotide hybridization reaction using thermodynamic method. Thermodynamic of 5 oligonucleotides with central and terminal mismatch sequences were obtained by measured UV-absorbance as a function of temperature. The data show that the nearest-neighbor base-pair model is adequate for predicting thermodynamics of oligonucleotides with average deviations for $\Delta$H$^{0}$ , $\Delta$S$^{0}$ , $\Delta$G$_{37}$ $^{0}$ and T$_{m}$, respectively.>$^{0}$ and T$_{m}$, respectively.

The Study of X Chromosome Inactivation Mechanism in Klinefelter's Syndrome by cDNA Microarray Experiment

  • Jeong, Yu-Mi;Chung, In-Hyuk;Park, Jung Hoon;Lee, Sook-Hwan;Chung, Tae-Gyu;Kim, Yong Sung;Kim, Nam-Soon;Yoo, Hyang-Sook;Lee, Suman
    • Genomics & Informatics
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    • v.2 no.1
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    • pp.30-35
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    • 2004
  • To investigate the XIST gene expression and its effect in a Klinefelter's patient, we used Klinefelter's syndrome (XXY) patient with azoospermia and also used a normal male (XY) and a normal female (XX) as the control, We were performed cytogenetic analysis, Y chromosomal microdeletion assay (Yq), semi-quantitative RT-PCR, and the Northern blot for Klinefelter's syndrome (KS) patient, a female and a male control, We extracted total RNA from the KS patient, and from the normal cells of the female and male control subjects using the RNA prep kit (Qiagen), cDNA microarray contained 218 human X chromosome-specific genes was fabricated. Each total RNA was reverse transcribed to the first strand cDNA and was labeled with Cy-3 and Cy-5 fluorescein, The microarray was scanned by ScanArray 4000XL system. XIST transcripts were detected from the Klinefelters patient and the female by RT-PCR and Northern blot analysis, but not from the normal male, In the cDNA microarray experiment, we found 24 genes and 14 genes are highly expressed in KS more than the normal male and females, respectively. We concluded that highly expressed genes in KS may be a resulted of the abnormal X inactivation mechanism.

Assessment of Relationship between Fyn-related Kinase Gene Polymorphisms and Overweight/Obesity in Korean Population

  • Jung, Mi-Young;Kim, Bum-Shik;Kim, Youn-Jung;Koh, In-Song;Chung, Joo-Ho
    • The Korean Journal of Physiology and Pharmacology
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    • v.12 no.2
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    • pp.83-87
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    • 2008
  • The fyn-related kinase (FRK) belongs to the tyrosine kinase family of protein kinases. Recent studies have shown that Frk affects pancreatic beta cell number during embryogenesis and promotes beta cell cytotoxic signals in response to streptozotocin. To investigate the genetic association between FRK polymorphisms and the risk of obesity in Korean population, single nucleotide polymorphisms (SNPs) in the FRK gene region were selected and analyzed. The body mass index (BMI) was calculated, and biochemical data (systolic blood pressure, diastolic blood pressure, hemoglobin A1C, triglyceride, total cholesterol, high density lipoprotein, and low density lipoprotein) of blood sample from each subject were also measured. One hundred fifty five healthy control and 204 overweight/obesity subjects were recruited. Genotype frequencies of six SNPs [rs6568920 (+8391G>A), rs3756772 (+56780A>G), rs3798234 (+75687C>T), rs9384970 (+68506G>A), rs1933739 (+72978G>A), and rs9400883 (+75809A>G)] in the FRK gene were determined by Affymetrix Targeted Genotyping Chip data. According to the classification of Korean Society for the Study of Obesity, control (BMI 18 to < 23) and overweight/obesity (BMI$\geq$23) subjects were recruited. For the analysis of genetic data, EM algorithm, SNPStats, Haploview, HapAnalyzer, SNPAnalyzer, and Helixtree programs were used. Multiple logistic regression analysis (codominant, dominant, and recessive models) was performed. Age and gender as covariates were adjusted. For biochemical data, Student's t test was used. The mean value of BMI in the control and overweigh/obesity groups was 21.1${\pm}$1.2 (mean${\pm}$SD) and 25.6${\pm}$2.0, respectively. All biochemical data of the overweight/obesity group were statistically significance, compared with the control group. Among six SNPs, two linkage disequilibrium (LD) blocks were discovered. One block consisted of rs1933739 and rs9400883, and the other comprised rs3756772 and rs3798234. One SNP (rs9384970, +68506G>A) showed an association with overweight/obesity in the codominant model (p=0.03). Interestingly, the AA genotype distribution in the overweight/obesity group (n=7, 3.5%) was higher than those in the control group (n=1, 0.6%), which is not found in either Japanese or Chinese subjects. Therefore, the AA genotype of rs9384970 may be a risk factor for development of obesity in Korean population. The results suggest that FRK may be associated with overweight/obesity in Korean population.

Ulcerative Colitis is Associated with Novel Polymorphisms in the Promoter Region of MIP-3${\alpha}$/CCL20 Gene

  • Choi, Suck-Chei;Lee, Eun-Kyung;Lee, Sung-Ga;Chae, Soo-Cheon;Lee, Myeung-Su;Seo, Geom-Seog;Kim, Sang-Wook;Yeom, Joo-Jin;Jun, Chang-Duk
    • IMMUNE NETWORK
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    • v.5 no.4
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    • pp.205-214
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    • 2005
  • Background: We examined global gene expression profiles of peripheral blood mononuclear cells (PBMCs) in patients with ulcerative colitis (DC), and tested whether the identified genes with the altered expression might be associated with susceptibility to UC. Methods: PBMCs from 8 UC and 8 normal healthy (NH) volunteers were collected, and total RNAs were subjected to the human 8.0K cDNA chip for the micro array analysis. Real time-PCR (RT-PCR) was performed to verify the results of micro array. One hundred forty UC patients and 300 NH controls were recruited for single nucleotide polymorphism (SNP) analysis. Results: Twenty-five immune function-related genes with over 2-fold expression were identified. Of these genes, two chemokines, namely, CXCL1 and CCL20, were selected because of their potential importance in the evocation of host innate and adaptive immunity. Four SNPs were identified in the promoter and coding regions of CXCL1, while there was no significant difference between all patients with UC and controls in their polymorphisms, except minor association at g.57A>G (rs2071425, p=0.02). On the other hand, among three novel and one known SNPs identified in the promoter region of CCL20, g. -1,706 G>A (p=0.000000055), g. -1,458 G>A (p=0.0048), and g. -962C>A (p=0.0006) were found to be significantly associated with the susceptibility of Uc. Conclusion: Altered gene expression in mononuclear cells may contribute to IBD pathogenesis. Although the findings need to be confirmed in other populations with larger numbers of patients, the current results demonstrated that polymorphisms in the promoter region of CCL20 are positively associated with the development of Uc.

Regulation of Pipernonaline on Biological Functions of Human Prostate Cancer Cells Based on Microarray Analysis (Microarray를 이용한 pipernonaline의 인간 전립선 암세포에 대한 기능 조절 분석)

  • Kim, Sang-Hun;Kim, Kwang-Youn;Yu, Sun-Nyoung;Park, Seul-Ki;Kwak, In-Seok;Rhee, Moon-Soo;Bang, Byung-Ho;Chun, Sung-Sik;Ahn, Soon-Cheol
    • Journal of Life Science
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    • v.22 no.11
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    • pp.1552-1557
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    • 2012
  • It has been reported that pipernonaline isolated from Piper longum Linn. has a wide biochemical and pharmacological effect, including antitumor activity in prostate cancer PC-3 cells. However, its mechanism and expression pattern of many genes involved in biological functions are not clearly understood. To perform the gene expression study in PC-3 cells treated with pipernonaline, a cDNA microarray chip composed of 44,000 human cDNA probes was used. As a result, cell cycle-related genes, apoptosis-related genes, and cell proliferation/growth-related genes have been identified in gene ontology of the DAVID database. These results suggest that pipernonaline has antitumor activity by regulating the expression pattern of genes involved in biological signaling pathway in prostate cancer PC-3 cells. Further, additional analysis of these microarray data can be a useful tool to identify the mechanism and discovery of novel genes in cancer therapy.

Microarray analysis of gene expression in raw cells treated with scolopendrae corpus herbal-acupuncture solution (蜈蚣(오공) 약침액(藥鍼液)이 LPS로 처리된 RAW 세포주(細胞柱)의 유전자(遺傳子) 발현(發顯)에 미치는 영향(影響))

  • Bae, Eun-Hee;Lee, Kyung-Min;Lee, Bong-Hyo;Lim, Seong-Chul;Jung, Tae-Young;Seo, Jung-Chul
    • Korean Journal of Acupuncture
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    • v.23 no.3
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    • pp.133-160
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    • 2006
  • Objectives : Scolopendrae Corpus has a broad array of clinical applications in Korean medicine, including treatment of inflammatory conditions such as arthritis. To explore the global gene expression profiles in human Raw cell lines treated with Scolopendrae Corpus herbal-acupuncture solution (SCHAS), cDNA microarray analysis was performed. Methods : The Raw 264.7 cells were treated with lipopolysaccharide (LPS), SCHAS, or both. The primary data was normalized by the total spots of intensity between two groups, and then normalized by the intensity ratio of reference genes such as housekeeping genes in both groups. The expression ratio was converted to log2 ratio. Normalized spot intensities were calculated into gene expression ratios between the control and treatment groups. Greater than 2 fold changes between two groups were considered to be of significance. Results : Of the 8 K genes profiled in this study, with a cut-off level of two-fold change in the expression, 20 genes (BCL2-related protein A1, MARCKS-like 1, etc.) were upregulated and 5 genes (activated RNA polymerase II transcription cofactor 4, calcium binding atopy-related autoantigen 1, etc.) downregulated following LPS treatment. 139 genes (kell blood group precursor (McLeod phenotype), ribosomal protein S7, etc.) were upregulated and 42 genes (anterior gradient 2 homolog (xenopus laevis), phosphodiesterase 8B, etc.) were downregulated following SCHAS treatment. And 10 genes (yeast saccharomyces cerevisiae intergeneic sequence 4-1, mitogen-activated protein kinase 1, etc.) were upregulated and 8 genes (spermatid perinuclear RNA binding protein, nuclear receptor binding protein 2, etc.) were downregulated following co-stimulation of SCHAS and LPS. Discussions : It is thought that microarrays will play an ever-growing role in the advance of our understanding of the pharmacological actions of SCHAS in the treatment of arthritis. But further studies are required to concretely prove the effectiveness of SCHAS.

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