• Title/Summary/Keyword: RNA microarray

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Gene Expression Profile in the Liver Tissue of High Fat Diet-Induced Obese Mice

  • Minho Cha;Bongjoo Kang;Kim, Kyungseon;Woongseop Sim;Hyunhee Oh;Yoosik Yoon
    • Nutritional Sciences
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    • v.7 no.1
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    • pp.8-16
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    • 2004
  • The purpose of this study was to investigate the gene profiles that were up- or down-regulated in the livers of high-fat diet-induced obese mice and $db_-/db_-$ mice with deficient leptin receptor. C57/BL6 normal mice and $db_-/db_-$ mice, respectively, were divided into two groups and fed a standard or high-fat diet for four weeks. Liver weight was unchanged in the normal mice but the high-fat diet led to a 10% weight increase in the $db_-/db_-$mice. Adipose tissue mass increased by about 88% in the normal mice that were fed a high-fat diet and by about 17% in the $db_-/db_-$mice on the high-fat diet. In terms of serum lipids, total cholesterol significantly increased in mice on the high-fat diet. Microarray analysis was carried out using total RNA isolated from the livers of standard or high-fat diet-fed mice of the normal and $db_-/db_-$ strains. The change of gene expression was confirmed by RT-PCR. About 1.6% and 6.8% of total genes, respectively, showed different expression patterns in the normal mice fed the high-fat diet and $db_-/db_-$ mice. As a result of microarray, many genes involved in metabolism and signal pathways were shown to have different expression patterns. Expression of Mgst3 gene increased in the livers of normal and $db_-/db_-$ mice that were fed a high-fat diet. Wnt7b and Ptk9l were down-regulated in the livers of the normal mice and $db_-/db_-$ mice that were fed a high-fat diet. In conclusion, a high-fat diet induced obesity and affected gene expression involved in metabolism and signal pathway.

Microarray Analysis of Gene Expression in Rat Hippocampus of Maternal Social Separation Model (모성 및 사회성 분리 백서 모델의 해마에서 유전자 칩을 이용한 유전자 발현 연구)

  • Lee, Hee Jae;Son, Chang Hee;Kwak, Hyong Ryol;Lee, Sang-Hyun;Han, Yoon Hee;Kim, Soo Young;Park, Jong-Ik;Chun, Wanjoo;Kim, Sung-Soo
    • Korean Journal of Biological Psychiatry
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    • v.13 no.2
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    • pp.110-116
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    • 2006
  • Objectives : Alteration of hippocampus was demonstrated in the maternal social separation(MSS) pups, separated from dams on postnatal day(pnd) 14 and placed alone. Therefore, to understand the molecular events involved in the MSS, we have initiated a search for gene profiles that are up or down-regulated in the hippocampus of MSS pups. Methods : Analysis of cDNA microarray was performed by using total RNA extracted from the hippocampus of control and MSS pups on pnd 17. Also, passive-avoidance test was demonstrated on pnd 35. Results : Up-regulation of Nedd4a was observed in the hippocampus of MSS pups. Also, MSS rats showed less elongation of latency in passive avoidance test. Conclusion : We suggest that environmental effects of MSS may be altered the neural and/or glial differentiation and synapse formation-related genes which may lead cognitive alterations in MSS rats.

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Whole Genomic Expression Analysis of Rat Liver Epithelial Cells in Response to Phenytoin

  • Kim, Ji-Hoon;Kim, Seung-Jun;Yeon, Jong-Pil;Yeom, Hye-Jung;Jung, Jin-Wook;Oh, Moon-Ju;Park, Joon-Suk;Kang, Kyung-Sun;Hwang, Seung-Yong
    • Molecular & Cellular Toxicology
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    • v.2 no.2
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    • pp.120-125
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    • 2006
  • Phenytoin is an anti-epileptic. It works by slowing down impulses in the brain that cause seizures. The recent microarray technology enables us to understand possible mechanisms of genes related to compounds which have toxicity in biological system. We have studied that the effect of a compound related to hepatotoxin in vitro system using a rat whole genome microarray. In this study, we have used a rat liver epithelial cell line WB-F344 and phenytoin as a hepatotoxin. WB-F344 was treated with phenytoin for 1 to 24 hours. Total RNA was isolated at times 1, 6 and 24h following treatment of phenytoin, and hybridized to the microarray containing about 22,000 rat genes. After analysis with clustering methods, we have identified a total of 1,455 differentially expressed genes during the time course. Interestingly, about 1,049 genes exhibited differential expression pattern in response to phenytoin in early time. Therefore, the identification of genes associated with phenytoin in early response may give important insights into various toxicogenomic studies in vitro system.

Screening of Ecotoxicant Responsive Genes and Expression Analysis of Benzo[a]pyrene-exposed Rockfish (Sebastes schlgeli)

  • Yum, Seung-Shic;Woo, Seon-Ock;Lee, Taek-Kyun
    • Molecular & Cellular Toxicology
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    • v.2 no.2
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    • pp.114-119
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    • 2006
  • Benzo[a]pyrene is a representative ecotoxicant in marine environment and a model compound of polycyclic aromatic hydrocarbons, which has an ability to bioaccumulate in aquatic organisms. This study aimed to identify molecular biomarkers suitable for assessing environmental pollution using a microarray technique. We examined the effects of benzo[a]pyrene on gene expressions in the rockfish, Sebastes schlegeli. We constructed the subtractive cDNA library with hepatic RNA from benzo[a]pyrene-exposed and non-exposed control fish. From the library 10,000 candidate clones were selected randomly and cDNA microarray was constructed. We determined benzo[a]pyrene-responsive genes using a high-density microarray. Statistical analysis showed that approximately 400 genes are significantly induced or reduced by benzo[a]pyrene treatment ($2\;{\mu}m$). Especially gene expression changes of 4 candidate clones among the up- or down-regulated genes were investigated in 6, 12 and 24 hr BaP-exposed fish groups. Many methods have been developed to monitor marine environmental status, which depend on quantifying the levels of the toxic components in polluted seawater or on ecological accessing, such as species diversity or richness. However, those methods could not provide information on physiological or genetic changes induced by such environmental stresses. Comparing with the conventional methods, these data will propose that benzo[a]pyrene-responsive genes can be useful for biological risk assessment of polycyclic aromatic hydrocarbons on marine organism at molecular level.

Gene Expression Profiling of Non-Hodgkin Lymphomas

  • Zekri, Abdel-Rahman Nabawy;Hassan, Zeinab Korany;Bahnassy, Abeer Ahmed;Eldahshan, Dina Hassan;El-Rouby, Mahmoud Nour Eldin;Kamel, Mahmoud Mohamed;Hafez, Mohamed Mahmoud
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.7
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    • pp.4393-4398
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    • 2013
  • Background: Chromosomal translocations are genetic aberrations associated with specific non-Hodgkin lymphoma (NHL) subtypes. This study investigated the differential gene expression profile of Egyptian NHL cases based on a microarray approach. Materials and Methods: The study included tissue samples from 40 NHL patients and 20 normal lymph nodes used as controls. Total RNA was extracted and used for cDNA microarray assays. The quantitative real time polymerase chain reaction was used to identify the aberrantly expressed genes in cancer. Results: Significant associations of 8 up-regulated and 4 down-regulated genes with NHL were observed. Aberrant expression of a new group of genes not reported previously was apparent, including down-regulated NAG14 protein, 3 beta hydroxy-delta 5-c27 steroid oxi-reductase, oxi-glutarate dehydrogenase (lipo-amide), immunoglobulin lambda like polypeptide 3, protein kinase x linked, Hmt1, and caveolin 2 Tetra protein. The up-regulated genes were Rb binding protein 5, DKFZP586J1624 protein, protein kinase inhibitor gamma, zinc finger protein 3, choline ethanolamine phospho-transferase CEPT1, protein phosphatase, and histone deacetylase-3. Conclusions: This study revealed that new differentially expressed genes that may be markers for NHL patients and individuals who are at high risk for cancer development.

GENE EXPRESSION AFTER THE APPLICATION OF THE FLUID-INDUCED SHEAR STRESS ON THE GINGIVAL FIBROBLAST (유체에 의해 유발된 전단력이 치은 섬유아세포 유전자 발현 변화에 미치는 영향에 관한 연구)

  • Jeong, Mi-Hyang;Choi, Je-Yong;Chae, Chang-Hoon;Kim, Seong-Gon;Nahm, Dong-Seok
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.27 no.5
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    • pp.424-430
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    • 2005
  • The oral cavity is humid environment mainly due to the continuous salivary flow. The reaction of oral mucosa to fluid flow is important for homeostasis and pathogenesis. The objective of this study is the screening the change of gene expression after the application of fluid induced shear stress (FISS) on the gingival fibroblast using cDNA microarray assay. The immortalized human gingival fibroblasts were grown and FISS was applied using a cone viscometer at a rotational velocity of 40 rpm, respectively for periods of 2 and 4 hours. The synthesis of cDNA was done from the extracted total RNA and cDNA microarray assay was done subsequently. The genes that showed over 1.6 in the Cy3/Cy5 or the Cy5/Cy3 value were regarded as genes influenced significantly by the FISS application ion (/M/>0.7). The " RUNX-1" was increased its expression in 2 hours group and " RUN and SH3 domain containing 1" was increased its expression in 4 hours group. The "CC020415", "cyclin L1", "interferon regulatory factor1", "early growth response 1", "immediate early response 2", and "immediate early response 3" genes were increased their expression in 2 and 4 hours after FISS application. In conclusion, we could find many genes that were probably related to the FISS application. Interestingly, most of them were placed in similar molecular pathways and these findings improve the reliability of chip data and usefulness in overall screening. From this experiment, we could find many items for further study and it will make improvement in the understanding of intracellular events in response to FISS.

cDNA Microarray Analysis of the Gene Expression Profile of Swine Muscle

  • Kim, Chul Wook;Chang, Kyu Tae;Hong, Yeon Hee;Jung, Won Yong;Kwon, Eun Jung;Cho, Kwang Keun;Chung, Ki Hwa;Kim, Byeong Woo;Lee, Jung Gyu;Yeo, Jung Sou;Kang, Yang Su;Joo, Young Kuk
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.8
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    • pp.1080-1087
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    • 2005
  • By screening specific genes related to the muscle growth of swine using cDNA microarray technology, a total of 5 novel genes (GF (growth factor) I, II, III, IV and V) were identified. Results of southern blotting to investigate the number of copies of these genes in the genome of swine indicated that GF I, GF III, and GF V existed as one copy and GF II, and GF IV existed as more than two copies. It was suggested that there are many isoforms of these genes in the genome of swine. Also, results of northern blotting to investigate whether these genes were expressed in grown muscle, using GF I, III, and V indicated that all the genes were much more expressed in the muscle of swine with body weight of 90 kg. Expression patterns of these genes in other organs, namely muscle and propagation and fat tissues, were investigated by extracting RNA from the tissues. These genes were not expressed in the propagation and fat tissues, but were expressed in the muscle tissue. To determine the mechanism of muscle growth, further studies should be preceded using the 3 specific genes related to muscle growth, that is GF I, III, and V.

DNA and Proteomic Analysis of Ginseng Radix Rubra Herbal-acupuncture Solution(GRR-HAS) on Gene Expression in HepG2 Carcinomar Cells (홍삼약침액(紅蔘藥鍼液)의 DNA와 단백질 발현(發顯)에 미치는 영향(影響))

  • Won, Eun-Ju;Lee, Bong-Hyo;Lim, Seong-Chul;Jung, Tae-Young;Seo, Jung-Chul;Lee, Kyung-Min
    • Journal of Acupuncture Research
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    • v.23 no.3
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    • pp.177-190
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    • 2006
  • Objectives : It has long been known about the anticancer effect of GRR-HAS, however, it has not been systemically determined the differentially regulated genes by GRR-HAS in cancer cells. The purpose of this study is to screen the GRR-HAS mediated differentially expressed genes in cancer cells such as HepG2 hepatoma cell lines. Oligonucleotide microarray and proteomic approaches were employed to screen the differential expression genes. Methods : GRR~HAS was prepared by boiling and stored at $-70^{\circ}C$ until use. Cells were treated with various concentrations of GRR-HAS (0.1, 0.5, 1.5, 10, $20mg/m{\ell}$) for 24 h. Cell toxicity was tested by MTT assay. To screen the differentially expressed genes in cancer cells, cells were treated with $1.5mg/m{\ell}$ of GRR-HAS. For oligonucleotide microarray assay, total RNA was used for gene expression analysis using oligonucleotide genechip (Human genome Ul33 Plus 2.0., Affimatrix Co.). For proteomic analysis, total protein was analyzed by 2D gel electrophoresis and Q-TOF mass spectrometer. Results : It has no cytotoxic effects on both HepG2 cells in all concentrations(0.1, 0.5, 1.5, 10,$20mg/m{\ell}$). In oligonucleotide microarray assay, the number of more than twofold differentially regulated known genes was 320 with 6 up-regulated and 314 down-regulated genes in HepG2 cells. In proteomic analysis, three spots were identified by 2D-gel electrophoresis and Q-TOF analysis. One down -regulated protein was protein disulfide isomerase and up-regulated proteins were fatty acid binding protein 1 and 14-3-3 gan1lTIa protein by $1.5mg/m{\ell}$ of CRR-HAS. Discussion : This study showed the comprehensive gene expression analysis using oligonucleotide microarray for the screening of GRR-HAS mediated differentially regulated genes. These results will provide a better application of GRR-HAS in cancer field and drug target development.

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GENE EXPRESSION ANALYSIS OF THE DENTAL PULP IN HEALTHY AND CARIES TEETH (치아 우식증에 따른 치수내 유전자 발현 변화에 관한 분석)

  • Oh, So-Hee;Kim, Jong-Soo
    • Journal of the korean academy of Pediatric Dentistry
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    • v.37 no.3
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    • pp.275-287
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    • 2010
  • Deep caries may induce pulpitis and the pulpal tissue interacts with microbial invasion. The immune response to protect the pulpal tissue can be mediated by cellular signal molecules produced by the pulpal cells. The understanding of these processes is important to find future therapeutic method for the diseased pulp. The pulp tissue from sound teeth was set as control group (n=30) and the pulp tissue from decayed teeth was set as test group (n=30). Total RNA was extracted from the pulp of each group and it was used for cDNA microarray and reverse transcriptase-polymerase chain reaction(RT-PCR). The expression of TGF-${\beta}1$ was studied by immunohistochemistry. The results were as follows: 1. cDNA microarray analysis identified 520 genes with 6-fold or greater difference in expression level with 143 genes more abundant in health and 377 genes more abundant in disease. 2. The RT-PCR analysis was done for randomly selected 14 genes and the results supported the result of cDNA microarray assay. 3. TGF-${\beta}1$ was highly expressed in the carious pulp and it was found in odontoblast by immunohistochemistry. In conclusion, many cytokines were found to be significantly changed their expression in the diseased pulp(/M/>1.6).

DNA and Proteomic Expression of Cervi parvum cornu Herbal-acupuncture Solution (CPC-HAS) in HepG2 carcinomar cells (녹용약침액(鹿茸藥鍼液)의 간암세포주(肝癌細胞柱)에 대한 DNA 및 단백질 발현(發顯))

  • Ryu, Sung-Hyun;Lee, Kyung-Min;Lee, Bong-Hyo;Lim, Seong-Chul;Jung, Tae-Young;Seo, Jung-Chul
    • Journal of Pharmacopuncture
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    • v.9 no.2
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    • pp.5-16
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    • 2006
  • Objective : It has long been known about the osteogenic effect of CPC-HAS 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 HepG2 hepatoma cells. Oligonucleotide microarray and proteomics approaches were 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, 20mg/ml) for 24 h. Cell toxicity was tested by MTT assay. To screen the differentially expressed genes in cancer cells, cells were treated with 1.5mg/ml of CPC-HAS. For oligonucleotide microarray assay, total RNA was used for gene expression analysis using oligonucleotide Genechip(Human genome Ul33 Plus 2.0., Affimatrix Co.). For proteomic analysis, total protein was analyzed by 2D gel electrophoresis and Q-TOF mass spectrometer. Results : It has no cytotoxic effects on both HepG2 cell in all concentrations(0.l, 0.5, 1.5, 10, 20mg/ml). In oligonucleotide microarray assay, the number of more than twofold differentially regulated known genes was 23 with 5 up-regulated and 18 down-regulated genes in HepG2 cells. In proteomic analysis, three spots were identified by 2D-gel electrophoresis and Q-TOF analysis. Two down-regulated proteins were aldehyde dehydrogenase 1 and enolase 1, and up-regulated protein was fatty acid binding protein 1 by 1.5mg/ml of CPC-HAS. Discussion : This study showed the screening of CPC-HAS mediated differentially regulated genes using combined approaches of oligonucleotide microarray and proteomic analysis. The screened genes will be used for the better understanding of the therapeutic effects of CPC-HAS on cancer fields.