• 제목/요약/키워드: PCR array

검색결과 109건 처리시간 0.029초

대규모 유전자 분석 기법을 이용한 육미지황원의 유전자 발현 연구 (Studies on Gene Expression of Yukmijihwang-tang using High-throughput Gene Expression Analysis Techniques)

  • 강봉주;김윤택;조동욱
    • 한국한의학연구원논문집
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    • 제8권2호통권9호
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    • pp.95-107
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    • 2002
  • Yukmijihwang-tang(YM) is a noted herbal prescription in Chinese and Korean traditional medicines, and it has been known to reinforce the vital essence and has been widely used for a variety of disease such as stroke, osteoporosis, anti-tumor, and hypothyrodism. Regarding its traditional use, YM has been known to reinforce the Yin (vital essence) of liver and kidney. Also it has been known to reinforce nutrition and biological function in brain. Recently, studies suggested that YM increase antioxidant activities and exert the protective effect against oxidant-induced liver cell injury. We investigated the high-throughput gene expression analysis on the Yukmijihwang-tang administrated in SD rats. Microarray data were validated on a limited number of genes by semiquantitative RT-PCR and Western blot analyses. The recent availability of microarrays provides an attractive strategy for elaborating an unbiased molecular profile of large number of genes in drug discovery This experimental approach offers the potential to identify molecules or cellular pathways not previously associated with herbal medicine. Total RNA from normal control brain and Yukmijihwang-tang administrated brain were hybridized to microarrays containing 10,000 rat genes. The 52 genes were found to be up-regulated(twice or more) excluding EST gene. The nine genes were found to be down-regulated(twice or more) excluding EST gene. Gene array technology was used to identify for the first time many genes expression pathway analysis that arecell cycle pathway, apoptosis pathway, electron transport chain pathway, cytoplasmic ribosomal protein pathway, fatty acid degradation pathway, and TGF-beta signaling pathway. These differentially expressed genes pathway analysis have not previously been iavestigated in the context of herbal medicine efficacy and represent novel factors for further study of the mechanism of herbal medicine efficacy.

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Targeting SHCBP1 Inhibits Cell Proliferation in Human Hepatocellular Carcinoma Cells

  • Tao, Han-Chuan;Wang, Hai-Xiao;Dai, Min;Gu, Cheng-Yu;Wang, Qun;Han, Ze-Guang;Cai, Bing
    • Asian Pacific Journal of Cancer Prevention
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    • 제14권10호
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    • pp.5645-5650
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    • 2013
  • Src homology 2 domain containing (SHC) is a proto-oncogene which mediates cell proliferation and carcinogenesis in human carcinomas. Here, the SHC SH2-domain binding protein 1 (SHCBP1) was first established to be up-regulated in human hepatocellular carcinoma (HCC) tissues by array-base comparative genome hybridization (aCGH). Meanwhile, we examine and verify it by quantitative real-time PCR and western blot. Our current data show that SHCBP1 was up-regulated in HCC tissues. Overexpression of SHCBP1 could significantly promote HCC cell proliferation, survival and colony formation in HCC cell lines. Furthermore, knockdown of SHCBP1 induced cell cycle delay and suppressed cell proliferation. Furthermore, SHCBP1 could regulate the expression of activate extracellular signal-regulated kinase 1/2 (ERK1/2) and cyclin D1. Together, our findings indicate that SHCBP1 may contribute to human hepatocellular carcinoma by promoting cell proliferation and may serve as a molecular target of cancer therapy.

E1/E2 of Hepatitis C Virus Genotype-4 and Apoptosis

  • Zekri, Abdel-Rahman N;Sobhy, Esraa;Hussein, Nehal;Ahmed, Ola S;Hussein, Amira;Shoman, Sahar;Soliman, Amira H;El-Din, Hanaa M Alam
    • Asian Pacific Journal of Cancer Prevention
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    • 제17권7호
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    • pp.3131-3138
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    • 2016
  • Several studies have addressed the possible role of hepatitis C virus genotype-4 (HCV GT4) in apoptosis. However, this still not fully understood. In the current study a re-constructed clone of E1/E2 polyprotein region of the HCV GT4 was transfected into the Huh7 cell line and a human apoptotic PCR array of 84 genes was used to investigate its possible significance for apoptosis. Out of the 84 genes, only 35 showed significant differential expression, 12 genes being up-regulated and 23 down-regulated. The highest-up regulated genes were APAF1 (apoptotic peptidase-activating factor 1), BID (BH3 interacting domain death agonist) and BCL 10 (B-cell CLL/lymphoma protein 10) with fold regulation of 33.2, 30.1 and 18.9, respectively. The most down-regulated were FAS (TNF receptor super family), TNFRSF10B (tumor necrosis factor receptor super-family member 10b) and FADD (FAS-associated death domain) with fold regulation of -30.2, -27.7 and -14.9, respectively. These results suggest that the E1/E2 proteins may be involved in HCV-induced pathogenesis by modulating apoptosis through the induction of the intrinsic apoptosis pathway and disruption of the BCL2 gene family.

Construction of a Bacterial Artificial Chromosome Library Containing Large BamHI Genomic Fragments from Medicago truncatula and Identification of Clones Linked to Hypernodulating Genes

  • Park So-Yeon;Nam Young-Woo
    • Journal of Microbiology and Biotechnology
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    • 제16권2호
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    • pp.256-263
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    • 2006
  • In the model legume Medicago truncatula, two mutants, sickle and sunn, exhibit morphologically and genetically distinct hypernodulation phenotypes. However, efforts to isolate the single recessive and single semidominant genes for sickle and sunn, respectively, by map-based cloning have so far been unsuccessful, partly due to the absence of clones that enable walks from linked marker positions. To help resolve these difficulties, a new bacterial artificial chromosome (BAC) library was constructed using BamHI-digested genomic fragments. A total of 23,808 clones were collected from ligation mixtures prepared with double-size-selected high-molecular-weight DNA. The average insert size was 116 kb based on an analysis of 88 randomly selected clones using NotI digestion and pulsed-field gel electrophoresis. About 18.5% of the library clones lacked inserts. The frequency of the BAC clones carrying chloroplast or mitochondrial DNA was 0.98% and 0.03%, respectively. The library represented approximately 4.9 haploid M. truncatula genomes. Hybridization of the BAC clone filters with a $C_{0}t-l$ DNA probe revealed that approximately 37% of the clones likely carried repetitive sequence-enriched DNA. An ordered array of pooled BAC DNA was screened by polymerase chain reactions using 13 sequence-characterized molecular markers that belonged to the eight linkage groups. Except for two markers, one to five positive BAC clones were obtained per marker. Accordingly, the sickle- and sunn-linked BAC clones identified herein will be useful for the isolation of these biotechnologically important genes. The new library will also provide clones that fill the gaps between preexisting BAC contigs, facilitating the physical mapping and genome sequencing of M. truncatula.

새우 양식장 주변 생물의 흰점바이러스 보유율 및 두토막눈썹참갯지렁이의 대하 Fenneropenaeus chinensis에 대한 흰점바이러스 전달 효과 (Prevalence of White Spot Syndrome Virus (WSSV) in Marine Organisms around the Shrimp Farm and Polychaete Worm-Mediated Transmission of WSSV to Fenneropenaeus chinensis)

  • 김근식;박상용;이일로;남윤권;방인철
    • 한국어병학회지
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    • 제22권1호
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    • pp.15-21
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    • 2009
  • White spot syndrome virus (WSSV) is a strong causative agent for high mortality in cultured and wild shrimps. From this study, the WSSV prevalence in marine organisms around shrimp farm as well as live feed-mediated transmission of WSSV to farmed shrimps were investigated. Based on nested-PCR method, WSSV was detected in wide array of marine organisms including Perinereis aibuhitensis (81.3% of prevalence rate, 13/16), Enedrias fangi (100%, 16/16), Ruditapes philippinarum (20%, 2/10), crab larvae (100%, 10/10), copepoda (30%, 3/10), Periophthalmus modestus (50%, 5/10), Pachygrapsus crassipes (10%, 1/10), Helice tridens (20%, 2/10) and Neomysis sp. (70%, 7/10). On the other hand, WSSV was not detected in Bullacta exarata, Uca arcuata, and Reishia clavigera. The percent prevalence of WSSV in wild shrimps, Fenneropenaeus chinensis was only 6%, but markedly increased up to 56% after a feeding trial using polychaete worms for one month, indicating that the live feed is one of significant carriers of WSSV to shrimps under practical farming conditions.

Specificity Protein 1 Expression Contributes to Bcl-w-Induced Aggressiveness in Glioblastoma Multiforme

  • Lee, Woo Sang;Kwon, Junhye;Yun, Dong Ho;Lee, Young Nam;Woo, Eun Young;Park, Myung-Jin;Lee, Jae-Seon;Han, Young-Hoon;Bae, In Hwa
    • Molecules and Cells
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    • 제37권1호
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    • pp.17-23
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    • 2014
  • We already had reported that Bcl-w promotes invasion or migration in gastric cancer cells and glioblastoma multiforme (GBM) by activating matrix metalloproteinase-2 (MMP-2) via specificity protein 1 (Sp1) or ${\beta}$-cateinin, respectively. High expression of Bcl-w also has been reported in GBM which is the most common malignant brain tumor and exhibits aggressive and invasive behavior. These reports propose that Bcl-w-induced signaling is strongly associated with aggressive characteristic of GBM. We demonstrated that Sp1 protein or mRNA expression is induced by Bcl-w using Western blotting or RT-PCR, respectively, and markedly elevated in high-grade glioma specimens compared with low-grade glioma tissues using tissue array. However, relationship between Bcl-w-related signaling and aggressive characteristic of GBM is poorly characterized. This study suggested that Bcl-w-induced Sp1 activation promoted expression of glioma stem-like cell markers, such as Musashi, Nanog, Oct4 and sox-2, as well as neurosphere formation and invasiveness, using western blotting, neurosphere formation assay, or invasion assay, culminating in their aggressive behavior. Therefore, Bcl-w-induced Sp1 activation is proposed as a putative marker for aggressiveness of GBM.

Bordetella bronchiseptica bateriophage suppresses B. bronchiseptica-induced inflammation in swine nasal turbinate cells

  • Park, Ga Young;Lee, Hye Min;Yu, Hyun Jin;Son, Jee Soo;Park, Sang Joon;Song, Kyoung Seob
    • Genes and Genomics
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    • 제40권12호
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    • pp.1383-1388
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    • 2018
  • The development of therapeutic bacteriophages will provide several benefits based on an understanding the basic physiological dynamics of phage and bacteria interactions for therapeutic use in light of the results of antibiotic abuse. However, studies on bacteriophage therapeutics against microbes are very limited, because of lack of phage stability and an incomplete understanding of the physiological intracellular mechanisms of phage. The major objective of this investigation was to provide opportunity for development of a novel therapeutic treatment to control respiratory diseases in swine. The cytokine array system was used to identify the secreted cytokines/chemokines after Bordetella bronchiseptica infection into swine nasal turbinate cells (PT-K75). We also performed the real-time quantitative PCR method to investigate the gene expression regulated by B. bronchiseptica infection or bacteriophage treatment. We found that B. bronchiseptica infection of PT-K75 induces secretion of many cytokines/chemokines to regulate airway inflammation. Of them, secretion and expression of IL-$1{\beta}$ and IL-6 are increased in a dose-dependent manner. Interestingly, membrane-bound mucin production via expression of the Muc1 gene is increased in B. bronchiseptica-infected PT-K75 cells. However, cytokine production and Muc1 gene expression are dramatically inhibited by treatment with a specific B. bronchiseptica bacteriophage (Bor-BRP-1). The regulation of cytokine profiles in B. bronchiseptica-induced inflammation by B. bronchiseptica bacteriophage is essential for avoiding inappropriate inflammatory responses. The ability of bacteriophages to downregulate the immune response by inhibiting bacterial infection emphasizes the possibility of bacteriophage-based therapies as a novel anti-inflammatory therapeutic strategy in swine respiratory tracts.

Ginsenoside Rg3 and Korean Red Ginseng extract epigenetically regulate the tumor-related long noncoding RNAs RFX3-AS1 and STXBP5-AS1

  • Ham, Juyeon;Jeong, Dawoon;Park, Sungbin;Kim, Hyeon Woo;Kim, Heejoo;Kim, Sun Jung
    • Journal of Ginseng Research
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    • 제43권4호
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    • pp.625-634
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    • 2019
  • Background: Ginsenoside Rg3, a derivative of steroidal saponins abundant in ginseng, has a range of effects on cancer cells, including anti-cell proliferation and anti-inflammation activity. Here, we investigate two long noncoding RNAs (lncRNAs), STXBP5-AS1 and RFX3-AS1, which are hypomethylated and hypermethylated in the promoter region by Rg3 in MCF-7 cancer cells. Methods: The lncRNAs epigenetically regulated by Rg3 were mined using methylation array analysis. The effect of the lncRNAs on the apoptosis and proliferation of MCF-7 cells was monitored in the presence of Rg3 or Korean Red Ginseng (KRG) extract after deregulating the lncRNAs. The expression of the lncRNAs and their target genes was examined using qPCR and Western blot analysis. The association between the expression of the target genes and the survival rate of breast cancer patients was analyzed using the Kaplan-Meier Plotter platform. Results: STXBP5-AS1 and RFX3-AS1 exhibited anti- and pro-proliferation effects, respectively, in the cancer cells, and the effects of Rg3 and KRG extract on apoptosis and cell proliferation were weakened after deregulating the lncRNAs. Of the genes located close to STXBP5-AS1 and RFX3-AS1 on the chromosome, STXBP5, GRM1, RFX3, and SLC1A1 were regulated by the lncRNAs on the RNA and protein level. Breast cancer patients that exhibited a higher expression of the target genes of the lncRNAs had a higher metastasis-free survival rate. Conclusion: The current study is the first to identify lncRNAs that are regulated by the presence of Rg3 and KRG extract and that subsequently contribute to inhibiting the proliferation of cancer cells.

Estimation and Classification of Flow Regimes for South Korean Streams and River

  • Park, Kyug Seo;Choi, Ji-Woong;Park, Chan-Seo;An, Kwang-Guk;Wiley, Michael J.
    • 한국수자원학회:학술대회논문집
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    • 한국수자원학회 2015년도 학술발표회
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    • pp.106-106
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    • 2015
  • The information of flow regimes continues to be norm in water resource and watershed management, in that stream flow regime is a crucial factor influencing water quality, geomorphology, and the community structure of stream biota. The objectives of this study were to estimate Korean stream flows from landscape variables, classify stream flow gages using hydraulic characteristics, and then apply these methods to ungaged biological monitoring sites for effective ecological assessment. Here I used a linear modeling approach (MLR, PCA, and PCR) to describe and predict seasonal flow statistics from landscape variables. MLR models were successfully built for a range of exceedance discharges and time frames (annual, January, May, July, and October), and these models explained a high degree of the observed variation with r squares ranging from 0.555 (Q95 in January) to 0.899 (Q05 in July). In validation testing, predicted and observed exceedance discharges were all significantly correlated (p<0.01) and for most models no significant difference was found between predicted and observed values (Paired samples T-test; p>0.05). I classified Korean stream flow regimes with respect to hydraulic and hydrologic regime into four categories: flashier and higher-powered (F-HP), flashier and lower-powered (F-LP), more stable and higher-powered (S-HP), and more stable and lower-powered (S-LP). These four categories of Korean streams were related to with the characteristics of environmental variables, such as catchment size, site slope, stream order, and land use patterns. I then applied the models at 684 ungaged biological sampling sites used in the National Aquatic Ecological Monitoring Program in order to classify them with respect to basic hydrologic characteristics and similarity to the government's array of hydrologic gauging stations. Flashier-lower powered sites appeared to be relatively over-represented and more stable-higher powered sites under-represented in the bioassessment data sets.

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파브리병의 바이오마커 발굴을 위한 파브리 마우스와 세포모델에서의 microRNA 발현 분석 (MicroRNA Expression Profiling in Cell and Mouse Models of Fabry Disease to Identify Biomarkers for Fabry Disease Nephropathy)

  • 정남희;박세영;전여진;최윤영;정성철
    • 대한유전성대사질환학회지
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    • 제15권3호
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    • pp.127-137
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    • 2015
  • 본 연구에서는 파브리병의 마우스 모델과 세포모델을 대상으로 miRNA expression microarray를 적용시켜 질환 모델과 정상 대조군 간의 전체 miRNA의 발현 차이를 조사하였고, 발현량에서 차이를 보인 특정 miRNA를 선별한 후, 해당 miRNA의 표적 유전자의 발현량 변화를 살펴보아 파브리병의 신장병변에 대한 바이오마커 발굴과 발병기전을 알아보고자 하였다. MicroRNA array 결과, 파브리 마우스 신장 조직의 경우, 1,247개의 분석 대상 miRNA 중 5개가 발현이 증가되어 있으며 3개가 발현이 감소되어 있음을 확인하였다. 그 중에서 miR-149-5p의 발현이 파브리 마우스의 신장에서 2배 이상 감소되어 있으며, 특히 35주령 이하의 파브리 마우스에서 이러한 감소현상이 나타남을 확인하였고, 또한 lyso-Gb3를 처리하여 배양한 SV40 MES 13 세포에서도 miR-149-5p의 발현이 감소됨을 알 수 있었다. miR-149-5p의 발현감소는 EMT와 관련된 유전자의 발현을 증가시킴을 확인하였다. 본 연구를 통해 miR-149-5p의 생체지표로서의 가능성과 함께 miR-149-5p의 발현감소가 EMT를 통한 파브리병에서의 사구체 섬유화에 관여할 것이라는 가능성을 제시하고 있다.