• 제목/요약/키워드: cDNA microarray chip

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

  • 김상헌;김광연;유선녕;박슬기;곽인석;이문수;방병호;전성식;안순철
    • 생명과학회지
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    • 제22권11호
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    • pp.1552-1557
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    • 2012
  • Pipernonaline은 후추나무과에 속하는 필발(Piper longum Linn.)의 유도체로서 전립선 암세포에 대한 항암활성이 보고되고 있다. 하지만 실제 암세포 내에서 생물학적 정보를 가진 수 많은 유전자들에 대한 발현이 어떻게 이루어지고 있는지 알려진 바가 없다. 본 연구에 사용된 microarray 분석은 동시에 수 만개 이상의 유전자 발현양상을 한번에 관찰할 수 있는 기술로서 특정 질병의 유전학적 특성과 기전 연구를 더 광범위하게 연구 할 수 있는 기술이다. 본 연구에서는 전립선 암세포인 PC-3 세포에 pipernonaline을 처리하여 cDNA microarray를 실시하였다. 이후, DAVID database를 이용하여 gene ontology의 Biological Process를 분석하여 세포사멸과 세포주기, 세포성장 및 증식에 관련된 유전자들을 우선적으로 분석하였다. 그 결과, 세포주기관련 256개, 세포사멸관련 197개, 세포성장 및 증식관련에 154개의 유전자가 확인 되었다. 이러한 결과는 pipernonaline은 전립선 암세포 내에 존재하는 생물학적 신호전달체계에 관련된 유전자 발현을 조절함으로써 항암활성을 나타내 것을 알 수 있었고, 이후 이러한 microarray의 추가적인 분석은 암세포 내 새로운 유전자의 탐색 및 메커니즘을 규명하는데 유용하게 사용할 수 있을 것으로 사료된다.

cDNA Microarray Analysis of Differential Gene Expression in Boar Testes during the Prepubertal Period

  • Lee, Dong-Mok;Lee, Ki-Ho;Choi, Jin Ho;Hyun, Jin Hee;Lee, Eun Ju;Bajracharya, Prati;Lee, Yong Seok;Chang, Jongsoo;Chung, Chung Soo;Choi, Inho
    • Asian-Australasian Journal of Animal Sciences
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    • 제22권8호
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    • pp.1091-1101
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    • 2009
  • In an attempt to understand the biochemical mechanism for the synthesis of the anabolic steroid, 19-nortestosterone, produced by prepubertal boar testes and its physiological role, normalized complementary DNA (cDNA) from boar testes was generated. DNA sequencing of 2,016 randomly selected clones yielded 794,116 base pairs of high quality nucleotide sequence. Computer-assisted assembly of the nucleotide sequence of each clone resulted in 423 contigs and 403 singletons including several genes for steroidogenic enzymes and molecules related to steroid metabolism. Analysis of gene expression pattern by use of the presently-fabricated cDNA microarray identified a number of genes that were differentially expressed during the postnatal development period in boar testes. Two genes of unknown function were identified to be highly expressed in the testis of 2-weeks-old neonatal boar. In addition, the sequencing of open reading frames of these genes revealed their homology with human alpha hemoglobin and Homo sapiens hypothetical LOC643669, transcript variant 1. Moreover, the transcripts of these genes were also detected in porcine muscle and adipocytes, in addition to Leydig cells of pigs.

방기(防己)의 투여가 비만 유발 쥐의 생리기능과 DNA Chip을 통한 유전자 발현에 미치는 영향에 대한 연구 (Effects of Sinomenium acutum Extract on Body Weight Gains and the DNA Chip Expression of Obese Rats.)

  • 조호근;김동일
    • 대한한방부인과학회지
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    • 제20권4호
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    • pp.41-55
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    • 2007
  • Purpose: This study is to examine anti-obesity effect and cytotoxicity of the long-term oral administration of Sinomenium acutum (Bang-gi, SA) Methods: Using diet-induced obesity C57BL/6 mouse model, anti-obesity effect and DNA chip expression and cytotoxicity of the long-term oral administration of this herbal extract were investigated. Results: The herbal extract treated groups were arrested in weight increment only when they were lodged together. Such effects were abolished when they kept individually. SA fed mice behaved very rudely and violently. On the basis of histological studies of liver tissues and also in vitro cytotoxicity tests of the liver and kidney cell lines, no significant toxicity was found by 14 weeks of SA treatments. However, we found significant changes in gene expression profile in SA treated group by micro-array analysis. In case of SA group, up-regulated genes were 1,213 and down-regulated ones were 2,558. Some of lipid metabolism related genes also significantly changed in both treatment groups. Conclusion: SA had effects of increasing the basal metabolic rate by stimulating the sympathetic nervous systems.

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마황(麻黃)의 투여가 비만 유발 쥐의 생리기능과 DNA Chip을 통한 유전자 발현에 미치는 영향에 대한 연구 (Effects of the oral administration of Epedra Sinica Extract on suppression of body weight gains and the DNA chip expression of obese rats.)

  • 조호근;양정민;김동일
    • 대한한방부인과학회지
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    • 제20권3호
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    • pp.65-80
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    • 2007
  • Purpose: This study is to examine anti-obesity effect and cytotoxicity of the long-term oral administration of Ephedra Sinica(Ma-hwang, ES) Methods: Using diet-induced obesity C57BL/6 mouse model, anti-obesity effect and DNA chip expression and cytotoxicity of the long-term oral administration of this herbal extract were investigated. Results: The herbal extract treated groups were arrested in weight increment only when they were lodged together. Such effects were abolished when they kept individually. ES fed mice behaved very rudely and violently. On the basis of histological studies of liver tissues and also in vitro cytotoxicity tests of the liver and kidney cell lines, no significant toxicity was found by 14 weeks of ES treatments. However, we found significant changes in gene expression profile in ES treated group by micro-array analysis. In case of ES group, up-regulated genes were 113 and down-regulated were 120. Some of lipid metabolism related genes also significantly changed in treatment groups. Conclusion: ES had effects of increasing the basal metabolic rate by stimulating the sympathetic nervous systems.

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인간 전립선암 PC-3 세포에서 Compound K에 의한 세포주기 조절 및 세포사멸 유전자 발현 변화 (Profile of Gene Expression Changes Treated with Compound K Induced Cell Cycle Arrest and Cell Death of Prostate Cancer PC-3 Cell Line)

  • 김광연;박광일;안순철
    • 대한한의학방제학회지
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    • 제29권4호
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    • pp.267-275
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    • 2021
  • Objectives : Previously, we reported that compound K isolated from fermented ginseng by Aspillus oryzae has a wide biochemical and pharmacological effect, including anti-cancer activity in prostate cancer PC-3 cells. Despite these findings, its signaling pathway and gene expression pattern are not clearly understood. Methods : To confirm the gene expression study of treated with compound K in PC-3 cells, a cDNA microarray chip composed of 44K human cDNA probes was used. MTT assay, western blot analysis, propidium iodide staining, and annexin V/propidium iodide staining were analyzed. Results : We confirmed the differences of gene expression profiles. Then, we analyzed with the cell cycle arrest, cell death and cell proliferation related genes using DAVID database. Conclusions : Our finding should be useful for understanding genome-wide expression patterns of compound K-mediated cell cycle arrest toward induction of cell death and be helpful for finding future cancer therapeutic targets for prostate cancer cells.

Bioinformatics for the Korean Functional Genomics Project

  • Kim, Sang-Soo
    • 한국생물정보학회:학술대회논문집
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    • 한국생물정보시스템생물학회 2000년도 International Symposium on Bioinformatics
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    • pp.45-52
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    • 2000
  • Genomic approach produces massive amount of data within a short time period, New high-throughput automatic sequencers can generate over a million nucleotide sequence information overnight. A typical DNA chip experiment produces tens of thousands expression information, not to mention the tens of megabyte image files, These data must be handled automatically by computer and stored in electronic database, Thus there is a need for systematic approach of data collection, processing, and analysis. DNA sequence information is translated into amino acid sequence and is analyzed for key motif related to its biological and/or biochemical function. Functional genomics will play a significant role in identifying novel drug targets and diagnostic markers for serious diseases. As an enabling technology for functional genomics, bioinformatics is in great need worldwide, In Korea, a new functional genomics project has been recently launched and it focuses on identi☞ing genes associated with cancers prevalent in Korea, namely gastric and hepatic cancers, This involves gene discovery by high throughput sequencing of cancer cDNA libraries, gene expression profiling by DNA microarray and proteomics, and SNP profiling in Korea patient population, Our bioinformatics team will support all these activities by collecting, processing and analyzing these data.

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자궁내막증 환자와 대조군에서의 자궁내막 유전자 발현의 차이: Microarray를 이용한 연구 (Comparison of Gene Expression Profile in Eutopic Endometria with or without Endometriosis: A Microarray Study)

  • 정민지;정은정;이신제;김문규;전상식;이택후
    • Clinical and Experimental Reproductive Medicine
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    • 제34권1호
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    • pp.19-31
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    • 2007
  • 목 적: 자궁내막증은 자궁내부에 존재하여야 할 자금내막조직이 자궁 외에 존재하는 질환으로 그 발생기전은 아직 명확하게 밝혀져 있지 않다. 이에 저자들은 자궁내막증 환자와 정상 대조군의 자궁내막조직 간의 유전자 발현의 차이가 자궁내막증의 발병과 관련이 있을 것이라는 가정 하에 DNA microarray 기술을 도입하여 연구를 시행하였다. 연구방법: 2002년 1월부터 2002년 12월까지의 기간 동안 본원 산부인과에서 자궁내막증 환자와 자궁내막증 이외의 다른 부인과적 질환으로 수술을 시행한 환자들을 대상으로 채취한 자궁내막 조직으로 KNU 4.8K cDNA chip을 이용하여 유전자 발현을 비교 연구하였다. 유전자칩으로 자궁내막증 조직에서 발현의 증감을 보였던 유전자 중에서 8종의 유전자를 대상으르 RT-PCR이나 real time RT-PCR 법을 통하여 그 발현 양상을 검증하였다. 결 과: 자궁내막증에 이환된 여성의 자궁내막조직에서 대조군에 비하여 높게 발현되고 있는 것으로 나타난 유전자들은 ATP synthase H transporting F1 (ATP5B), eukaryotic translation elongation factor 1, isocitrate dehydrogenase 1 (NADP+), mitochondrial ribosomal protein L3, ATP synthase H+ trarsporting (ATP5C1), LPS induced TNF-$\alpha$ factor 등으로 세포의 에너지 생성과 대사과정 및 신호전달에 관여하는 유전자들이었다. 한편 자궁내막중 환자의 자궁내막조직에서 대조군에 비하여 낮게 발현된 유전자들은 insulin like growth factor II associated protein, EGF-containing fibulin-like EMP1, matrix Gla protein, TGF beta-induced, TGF beta receptor 1(activin A receptor type II-like kinase), cystallin alpha B, fibulin 5, tissue inhibitor of metalloproteinase 3, collage type XII, alpha 1, tissue inhibitor of metalloproteinase 1, decorin 등으로 세포외기질의 구성 및 기능에 관련이 있었다. 결 론: 이상의 DNA mirroarry 및 RT-PCR을 통해 얻어진 결과에서 자궁내막증의 자궁내막조직에서 대조군에 비하여 유전자들의 발현에 차이가 있음을 확인하였다.

Array-based Nano-amplification Technique Was Applied in Detection of Hepatitis E Virus

  • Liu, Hui-Hui;Cao, Xuan;Yang, Yong;Liu, Ming-Gui;Wang, Ye-Fu
    • BMB Reports
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    • 제39권3호
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    • pp.247-252
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    • 2006
  • A rapid method for the detection of Hepatitis E Virus (HEV) was developed by utilizing nano-gold labeled oligonucleotide probes, silver stain enhancement and the microarray technique. The 5'-end -$NH_2$ modified oligonucleotide probes were immobilized on the surface of the chip base as the capture probe. The detection probe was made of the 3'-end -SH modified oligonucleotide probe and nano-gold colloid. The optimal concentrations of these two probes were determined. To test the detection sensitivity and specificity of this technique, a conservative fragment of the virus RNA was amplified by the RT-PCR/PCR one step amplification. The cDNA was hybridized with the capture probes and the detection probes on microarray. The detection signal was amplified by silver stain enhancement and could be identified by naked eyes. 100 fM of amplicon could be detected out on the microarray. As the results, preparation of nano-gold was improved and faster. Development time also was shortened to 2 min. Thus, considering high efficiency, low cost, good specificity and high sensitivity, this technique is alternative for the detection of HEV.

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

  • 유대은;강미선;박은정;김일찬;이재성;박광식
    • Environmental Analysis Health and Toxicology
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    • 제20권2호
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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.

cDNA Microarray Analysis of Phytophthora Resistance Related Genes Isolated from Pepper

  • Kim, Hyounjoung;Lee, Mi-Yeon;Kim, Ukjo;Lee, Sanghyeob;Park, Soon-Ho;Her, Nam-Han;Lee, Jing-Ha;Yang, Seung-Gyun;Harn, Chee-Hark
    • 한국식물병리학회:학술대회논문집
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    • 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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    • pp.67.1-67
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    • 2003
  • Phytophthora blight is a devastating disease of pepper and occurs almost anywhere peppers are grown. Phytophthora blight is caused by Phytophthora capsici and this pathogen can infect every part of the plant by moving inoculum in the soil, by infecting water on surface, by aerial dispersal to sporulating lesions. Management of Phytophthora blight currently relies on cultural practices, crop rotation, and use of selective fungicides. Since these treatments are a short-term management, a classical breeding for development of resistant pepper against the Phytophthora is an alternative. So far some of the resistant cultivars have been on the market, but those are limited regionally and commercially. Therefore, ultimately an elite line resistant against this disease should be developed, if possible, by biotechnology. We have set out a series of work recently in order to develop Phytophthora resistant pepper cultivar. For the first time, the cDNA microarray analysis was peformed using an EST chip that holds around 5000 pepper EST clones to identify genes responsive to Phytophthora infection. Total RNA samples were obtained from Capsicum annuum PI201234 after inoculating P. capsici to roots and soil and exposed to the chip. .Around 900 EST clones were up-regulated and down-regulated depending on the two RNA sample tissues, leaf and root. From those, we have found 55 transcription factors that may be involved in gene regulation of the disease defense mechanism. Further and in detail information will be provided in the poster.

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