• 제목/요약/키워드: Microarray assay

검색결과 122건 처리시간 0.031초

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

  • 오소희;김종수
    • 대한소아치과학회지
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    • 제37권3호
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    • pp.275-287
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    • 2010
  • 치아 우식증은 범발성 질환이고 이에 대한 생체반응은 단순하지 않으며 질병 과정과 숙주의 활성 모두를 반영하는 복합적인 반응이다. 이러한 반응을 이해하기 위해서는 질병의 세포학적, 분자학적인 면을 이해하는 것이 필수적이다. 이에 본 연구는 임상적으로 건강한 치아와 우식이 진행된 치아로부터 얻어진 치수 안의 유전자 발현을 규명하고 우식 병소에서 일어나는 치유 및 재생에 관계되는 분자와 면역 세포들 사이의 분자 생화학적 상호작용을 규명하기 위해서 우식치아와 건전치아의 치수를 이용하여 cDNA 미세배열(microarray) 분석과 역전사효소 중합효소 연쇄반응 (RT-PCR) 분석, 그리고 면역화학염색법 (immunohistochemistry)을 시행하여 다음과 같은 결론을 얻었다. 1. cDNA 미세배열 분석 결과, 건전치아군인 대조군에서는 143개의 유전자가, 우식치아군인 실험군에서는 377개의 유전자가 1.6배이상 발현되었다. 2. 역전사효소 중합효소 연쇄반응 분석에서 14개의 유전자를 선택하였고 cDNA 미세배열 분석결과와 동일한 결과를 확인하였다. 3. TGF-${\beta}1$의 면역조직화학적 관찰 결과, 건전치에 비해 우식치의 상아모세포와 치수에서 특히 강하게 발현되었다.

Oligonucleotide chip를 이용한 홍화자약침액(紅花子藥鍼液)이 간암세포주(肝癌細胞柱)의 유전자(遺傳子) 발현(發顯)에 미치는 영향(影響) (Effect of Carthami Tinctorii Fructus Herbal-acupuncture Solution(CTF-HAS) on Gene Expression in HepG2 carcinomar cells)

  • 이경민;임성철;정태영;서정철;한상원
    • Journal of Acupuncture Research
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    • 제22권3호
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    • pp.215-225
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    • 2005
  • 홍화자약침액(紅花子藥鍼液)의 항암효능(抗癌效能)을 밝히고자 간암세포주(肝癌細胞柱)에 최신 oligonucleotide chip assay 법을 통하여 대양(大量)의 유전자(遺傳子) 발현(發顯)을 분석(分析)한 결과(結果) 다음과 같은 결론(結論)을 얻었다. 1. MTT 분석(分析)에서 간암(肝癌) 및 위암세포주(胃癌細胞柱)는 홍화자약침액(紅花子藥鍼液) 1.5, 10, 20m/$m{\ell}$에서 대조군(對照群)에 비해 유의(有意)한 세포활성(細胞活性) 감소(減少)를 보였다. 2. 간암세포주(肝癌細胞柱)에 홍화자약침액(紅花子藥鍼液) 처치(處置) 시(時) 대조군(對照群)에 비해 발현(發顯)이 2배 이상 증가(增加)된 유전자(遺傳子)는 UCIA PMARARA fusion protein, human oral cancer candidate gene mRNA, EPPIN-3 등 19개였다. 3. 간암세포주(肝癌細胞柱)에 홍화자약침액(紅花子藥鍼液) 처치(處置) 시 (時)대조군(對照群)에 비해 유전자(遺傳子)의 발현(發顯)이 2배 이상 감소(減少)된 것은 BTF3, MMP11, paxillin, villinn 등 13개였다. 이상과 같이 홍화자약침액(紅花子藥鍼液)에 대한 간암세포주(肝癌細胞柱)의 유전자(遺傳子) 발현(發顯)을 oligonucleotide 분석(分析)으로 대량(大量) 검소(儉素)할 수 있었고, 심한 발현(發顯) 차이(差異)를 나타내는 각 유전자(遺傳子)는 암화(癌化) 과정(過程)이나, 홍화자약침액(紅花子藥鍼液)에 반응하는 유전자(遺傳子)로 치료제 개발을 위한 기초 자료로 활용할 수 있을 것으로 사료(思料)된다.

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신기술 이용 식중독균 신속검출법 개발 동향 분석 (Trends in Rapid Detection Methods for Food-borne pathogenic Microorganisms by Using New Technologies)

  • 김현주;김용수;정명섭;오덕환;전향숙;하상도
    • 한국식품위생안전성학회지
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    • 제25권4호
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    • pp.376-387
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    • 2010
  • 식품으로부터 식중독세균을 검출하는 대부분의 전통적인 방법들은 분리하고자 하는 미생물에 적합한 선택배지에 의존할 수밖에 없다. 이러한 선택배지들은 높은 경제성에도 불구하고 오래 걸려 때를 놓치게 되는 소요 시간이 항상 걸림돌로 작용하고 있으며, 하나의 선택배지로 한의 균만을 선택적으로 분리해 내는 등의 단점이 있다. 이러한 단점을 최소화하기 위하여 IMS 등 면역학적 또는 분자생물학적 기법들을 접목시키고 있고, 증균에서 분리와 확인을 동시에 할 수 있는 package 형태의 제품들이 시판되고 있다. 또한 이러한 제품 중 일부는 국제표준기구로부터 인증을 받았고 대부분 chromogenic 기질을 함유한 선택배지들이 포함되어 있을 만큼 신뢰도가 높다고 할 수 있다. 그러나 선택배지 자체만은 이들 단점을 해결할 수 없어 항체를 활용한 신속, 간편하고 환경친화적인 ELISA법, 검출장비가 불필요하여 가장 신속, 간편하게 현장에서 사용하도록 기술을 향상시킨 paper-strip kit나 ELISA 분석 시 직면하는 시료의 matrix effect를 최소화할 수 있다는 장점을 지닌 형광면역분석법(fluoroimmunoassay)등 새로운 기술이 용도에 맞게 다양하게 개발되고 있다. 또한 동시진단 능력이 우수한 현장 진단형 시스템이 개발될 경우, 향후 신 시장 창출, 바이오칩 기술발전, 식중독균을 포함한 위해 미생물의 동시진단 및 역학조사 등에 지대한 기여를 할 것으로 판단된다. 식품에서 식품위해미생물 검출에 있어서 가장 이상적인 방법은 검출 과정이 간단하고 경제적이며 정확하여야 한다는 것이다. 또한 정량적 결과 산출이 가능하여야 하고, 자동화된 방법으로 전환이 가능하여야 산업에 적용될 수 있다는 것인데, 이러한 이상적인 검출방법은 현재까지 존재하지 않는다. 다만, 여러 방법들이 각각의 장점을 갖고 있을 뿐이다. 즉, 현실적 대안은 현재까지 개발된 각각의 방법을 목적과 대상에 따라 시의적절하게 병용 사용하여야 한다는 것이다.

cDNA Microarray를 이용한 구강편평세포암종 세포주에서 $Taxol^{(R)}$과 Cyclosporin A로 유도된 유전자 발현양상 (GENE EXPRESSION PATTERNS INDUCED BY $TAXOL^{(R)}$ AND CYCLOSPORIN A IN ORAL SQUAMOUS CELL CARCINOMA CELL LINE USING CDNA MICROARRAY)

  • 김용관;이재훈;김철환
    • Maxillofacial Plastic and Reconstructive Surgery
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    • 제28권3호
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    • pp.202-212
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    • 2006
  • It is well-known that paclitaxel($Taxol^{(R)}$), which is extracted from the pacific and English yew, has been used as a chemotherapeutic agent for ovarian carcinoma and advanced breast carcinoma and Cyclosporin A, which is highly lipophilic cyclic peptide and isolated from a fungus, has been also used as an useful immunosuppressive drug after transplantation and is associated with cellular apoptosis. Since 1953, in which James Watson, Rosalind Franklin and Francis Crick discovered the double helical structure of DNA, a few kinds of techniques for identifying gene expression have been developed. In postgenomic period, many of researchers have used the DNA microarray which is high throughput screening technique to screen large numbers of gene expression simultaneously. In this study, we searched and screened the gene expression in the oral squamous cell carcinoma cell lines treated with $Taxol^{(R)}$, cyclosporin or cyclosporin combined with $Taxol^{(R)}$ using cDNA microarray. The results were as following; 1. It was useful that the appropriate concentration of Cyclosporin A and $Taxol^{(R)}$ used in oral squamous cell carcinoma cell line was under 1${\mu}g/ml$ and 3${\mu}g/ml$. 2. In the experimental group in which $Taxol^{(R)}$ and $Taxol^{(R)}$ + Cyclosporin A were used, the cell growth was extremely decreased. 3. In the group in which Cyclosporin A was used, the MTT assay was rarely decreased which means the activity of succinyl dehydrogenase is remained in mitochondria but in the group in which the mixture of Cyclosporin A and $Taxol^{(R)}$ were used, the MTT assay was extremely decreased. 4. In the each group in which Cyclosporin A(3 ${\mu}g/ml$) and $Taxol^{(R)}$(1 ${\mu}g/ml$) were used, the cell arrest was appeared in $G_2/M$ phase and in the group in which $Taxol^{(R)}$(3 ${\mu}g/ml$) was used, the cell arrest was appeared in both S phase and $G_2/M$ phase. 5. In the oral squamous cell carcinoma cell line treated with $Taxol^{(R)}$, several genes including ANGPTL4, RALBP1 and TXNRD1, associated with apoptosis, SUI1, MAC30, RRAGA and CTGF, related with cell growth, HUS1 and DUSP5, related with cell cycle and proliferation, ATF4 and CEBPG, associated with transcription factor, BTG1 and VEGF, associated with angiogenesis, FDPS, FCER1G, GPA33 and EPHA4 associated with signal transduction and receptor activity and AKR1C2 and UGTA10 related with carcinogenesis were detected in increased levels. The genes that showed increaced expression in the oral squamous cell carcinoma cell line treated with Cyclosporin A were CYR61, SERPINB2, SSR3 and UPA3A which are known as genes associated with cell growth, carcinogenesis, receptor activity and transcription factor. The genes expressed in the HN22 cell line treated with cyclosporin combined with $taxol^{(R)}$ were ALCAM and GTSE1 associated with cancer invasiveness and cell cycle regulation.

Microarray를 이용한 작약(芍藥)의 인간치은섬유모세포 유전자 발현 조절 연구 (Gene expression microarray analysis of Paeoniae radix on IL-$1{\beta}$-stimulated primary human gingival fibroblast)

  • 김경호;최영곤;홍연미;여수정;최지훈;김영홍;이제현;임사비나
    • 대한한의학회지
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    • 제31권2호
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    • pp.91-108
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    • 2010
  • Background & Objective: The aim of this study was to investigate the effect of P. radix on the inflammatory related gene expression in IL-$1{\beta}$-stimulated primary human gingival fibroblast using Whole Transcript Sense Target (WT-ST). Method: Human gingival fibroblast was incubated with P. radix [100 or $200\;{\mu}g/ml$], and IL-$1{\beta}$ [$1ng/m{\ell}$] added an hour later. After 24h, total RNA was extracted using RNeasy Mini Kit and the whole gene expression patterns were performed using WT-ST Labeling $Assay^{(R)}$. Result: In the DEG results, 782 genes were up-regulated in the IL-$1{\beta}$-treated group as compared to control and among those, 43 genes were associated with inflammation. 981 genes were down-regulated after treatment with IL-$1{\beta}$ and of those 7 genes were associated with inflammation. 1439 genes were up-regulated after treatment with P. radix plus IL-$1{\beta}$-treated when compared to IL-$1{\beta}$-treated alone group and 1225 genes were down-regulated in the same condition. Among the down-regulated genes, 5 were associated with inflammation- and inhibitor genes such as GDF15 and LIF. In the analysis of the P. radix plus IL-$1{\beta}$-treated group, the most significant pathways were the cytokine-cytokine receptor interaction, toll-like receptor signaling, JAK-STAT signaling and tyrosine metabolism. The gene expression patterns in the P. radix $200{\mu}g/m{\ell}$ plus IL-$1{\beta}$-treated group appear to be more involved in the metabolism-related pathways than in the $100{\mu}g/m{\ell}$ plus IL-$1{\beta}$-treated group. Conclusion & Discussion: By microarray analysis of gene expression data, we are able to identify gene expression patterns associated with not only anti-inflammation effect but also transcription function of P. radix.

인간 대장암 세포주에서 파이토케미칼 처리에 의한 유전자 발현 변화 (Global Gene Expression Changes by Several Phytochemicals in Human Colorectal Cancer Cell)

  • 박민희;곽은희;손호용;;김종식
    • 생명과학회지
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    • 제21권9호
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    • pp.1219-1225
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    • 2011
  • 파이토케미칼은 식물유래의 비 영양 성분으로서 항염증, 항산화, 및 화학적 암 예방 등의 생리활성을 가지고 있는 물질이다. 본 연구에서, 우리는 다섯 가지의 다른 파이토케미칼(resveratrol, genistein, epicatechin gallate, diallyl disulfide, caffeic acid phenethyl ester)이 대장암 세포주의 성장과 유전자 발현에 미치는 영향을 연구하였다. 세포 생존율 연구결과, 처리한 ECG를 제외한 모든 파이토케미칼에 의해 농도의존적으로 세포생존율이 감소함을 확인하였다. 또한, oligo DNA microarray 실험을 통해 다섯 종류의 파이토케미칼에 의해 공통적으로 증가 되는 유전자 6개와 공통적으로 발현이 감소되는 유전자 7개를 선별하였다. 공통적으로 발현이 증가되는 유전자를 선택하여 RT-PCR 방법을 통해 발현을 증명하였다. 또한, 파이토케미칼에 의한 NAG-1 단백질의 발현 증가도 확인하였다. 이러한 연구결과는 파이토케미칼에 의해 중재되어 지는 화학적 암 예방법의 일반적인 분자 기전을 이해하는데 도움을 줄 것으로 생각된다.

In silica Prediction of Angiogenesis-related Genes in Human Hepatocellular Carcinoma

  • Kang, Seung-Hui;Park, Jeong-Ae;Hong, Soon-Sun;Kim, Kyu-Won
    • Genomics & Informatics
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    • 제2권3호
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    • pp.134-141
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    • 2004
  • Hepatocellular carcinoma (HCC) is one of the most common malignancies worldwide and a typical hypervascular tumor. Therefore, it is important to find factors related to angiogenesis in the process of HCC malignancy. In order to find angiogenesis-related factors in HCC, we used combined methods of in silico prediction and an experimental assay. We analyzed 1457 genes extracted from cDNA microarray of HCC patients by text-mining, sequence similarity search and domain analysis. As a result, we predicted that 16 genes were likely to be involved in angiogenesis and then the effects of these genes were confirmed by hypoxia response element(HRE)-luciferase assay. For instant, we classified osteopontin into a potent angiogenic factor and coagulation factor XII into a significant anti­angiogenic factor. Collectively, we suggest that using a combination of in silico prediction and experimental approaches, we can identify HCC-specific angiogenesis­related factors effectively and rapidly.

Aptamers as Functional Nucleic Acids: in vitro Selection and Biotechnological Applications

  • You, Kyung-Man;Lee, Sang-Hyun;Aesul Im;Lee, Sun-Bok
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제8권2호
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    • pp.64-75
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    • 2003
  • Aptamers are functional nucleic acids that can specially bind to proteins, peptides, amino acids. nucleotides, drugs, vitamins and other organic and inorganic compounds. The aptamers are identified from random DNA or RNA libraries by a SELEX (systematic evolution of ligands by exponential amplification) process. As aptamers have the advantage, and potential ability to be released from the limitations of antibodies, they are attractive to a wide range of therapeutic and diagnostic applications. Aptamers, with a high-affinity and specificity, could fulfil molecular the recognition needs of various fields in biotechnology. In this work, we reviewed some aptamer Selection techniques, properties, medical applications of their molecules and their biotechnological applications, such as ELONA (enzyme linked oligonucleotide assay), flow cytometry, biosensors, electrophoresis, chromatography and microarrays.

Protein Microarrays and Their Applications

  • Lee, Bum-Hwan;Teruyuki Nagamune
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제9권2호
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    • pp.69-75
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    • 2004
  • In recent years, the importance of proteomic works, such as protein expression, detection and identification, has grown in the fields of proteomic and diagnostic research. This is because complete genome sequences of humans, and other organisms, progress as cellular processing and controlling are performed by proteins as well as DNA or RNA. However, conventional I protein analyses are time-consuming; therefore, high throughput protein analysis methods, which allow fast, direct and quantitative detection, are needed. These are so-called protein microarrays or protein chips, which have been developed to fulfill the need for high-throughput protein analyses. Although protein arrays are still in their infancy, technical development in immobilizing proteins in their native conformation on arrays, and the development of more sensitive detection methods, will facilitate the rapid deployment of protein arrays as high-throughput protein assay tools in proteomics and diagnostics. This review summarizes the basic technologies that are needed in the fabrication of protein arrays and their recent applications.

Toxicogenomics and Cell-based Assays for Toxicology

  • Tong, Weida;Fang, Hong;Mendrick, Donna
    • Interdisciplinary Bio Central
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    • 제1권3호
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    • pp.10.1-10.5
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    • 2009
  • Toxicity is usually investigated using a set of standardized animal-based studies which, unfortunately, fail to detect all compounds that induce human adverse events and do not provide detailed mechanistic information of observed toxicity. As an alternative to conventional toxicology, toxicogenomics takes advantage of currently advanced technologies in genomics, proteomics, metabolomics, and bioinformatics to gain a molecular level understanding of toxicity and to enhance the predictive power of toxicity testing in drug development and risk/safety assessment. In addition, there has been a renewed interest, particularly in various government agencies, to prioritize and/or supplement animal testing with a battery of mechanistically informative in vitro assays. This article provides a brief summary of the issues, challenges and lessons learned in these fields and discuss the ways forward to further advance toxicology using these technologies.