• Pediatric Gastroenterology, Hepatology & Nutrition
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• 제8권2호
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• pp.177-193
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• 2005

목 적: 담즙정체를 보이는 환자의 간조직에서 cDNA microarray를 이용한 유전자 발현 분석을 통 하여 담도 폐쇄증의 발생과정에 관여하는 유전자를 규명하고자 연구를 시행하였다. 방 법: 환자의 간조직은 담즙정체를 보이는 11명의 환자로부터 진단적 간생검시 얻은 간조직의 일부를 이용하였다. 이들의 원인 질환은 담도 폐쇄증이 7명, 신생아 간염 증후군이 4명이었다. 정상대조군의 간조직은 생체부분 간이식시 성인 공여자로부터 얻은 간조직의 일부를 이용하였다. 간조직에서 분리된 mRNA를 4.7K 인간유전자 cDNA microarray를 이용하여 유전자 발현의 양상을 분석하였다. 결 과: 분석한 4,700종의 유전자 중 담즙정체 환자 11명 모두의 간조직에서 발현이 증가된 유전자는 17종이 발견되었고, 발현이 감소된 유전자는 20종이 발견되었다. 담도 폐쇄증 환자와 신생아 간염 증후군 환자 사이에 49종의 유전자에서 발현의 차이가 관찰되었고, 특히 담도 폐쇄증에서는 발현이 증가하였으나 신생아 간염 증후군에서는 발현이 감소된 유전자는 24종이었으며 이들 유전자 발현의 차이에 의해서 두 군간의 감별이 가능하였다. 결 론: 신생아 간질환에서 담도 폐쇄증 환자들은 모두 일정한 양상을 보여주어 이에 대한 분석을 통하여 담도 폐쇄증의 조기 감별 진단을 할 수 있을것을 시사하였다. 또한 담도 폐쇄증에서 특이 발현을 보이는 유전자의 추가적인 기능 연구를 시행함으로써 담도 폐쇄증의 원인을 규명하는데 도움을 줄 수 있을 것으로 생각한다.

• Asian Pacific Journal of Cancer Prevention
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• 제13권7호
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• pp.3165-3172
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• 2012

Objective: Genetic polymorphisms in metabolic enzymes are associated with numerous cancers. A large number of single nucleotide polymorphisms (SNPs) in the CYP2D6 gene have been reported to associate with cancer susceptibility. However, the results are controversial. The aim of this Human Genome Epidemiology (HuGE) review and meta-analysis was to summarize the evidence for associations. Methods: Studies focusing on the relationship between CYP2D6 gene polymorphisms and susceptibility to cancer were selected from the Pubmed, Cochrane library, Embase, Web of Science, Springerlink, CNKI and CBM databases. Data were extracted by two independent reviewers and the meta-analysis was performed with Review Manager Version 5.1.6 and STATA Version 12.0 software. Odds ratios (ORs) with 95% confidence intervals (95%CIs) were calculated. Results: According to the inclusion criteria, forty-three studies with a total of 7,009 cancer cases and 9,646 healthy controls, were included in the meta-analysis. The results showed that there was a positive association between heterozygote (GC) of rs1135840 and cancer risk (OR=1.92, 95%CI: 1.14-3.21, P=0.01). In addition, we found that homozygote (CC) of rs1135840 might be a protective factor for cancer (OR=0.58, 95%CI: 0.34-0.97, P=0.04). Similarly, the G allele and G carrier (AG + GG) of rs16947 and heterozygote (A/del) of rs35742686 had negative associations with cancer risk (OR=0.69, 95%CI: 0.48-0.99, P=0.04; OR=0.60, 95%CI: 0.38-0.94, P=0.03; OR=0.50, 95%CI: 0.26-0.95, P=0.03; respectively). Conclusion: This meta-analysis suggests that CYP2D6 gene polymorphisms are involved in the pathogenesis of various cancers. The heterozygote (GC) of rs1135840 in CYP2D6 gene might increase the risk while the homozygote (CC) of rs1135840, G allele and G carrier (AG + GG) of rs16947 and heterozygote (A/del) of rs35742686 might be protective factors.

• Asian Pacific Journal of Cancer Prevention
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• 제15권20호
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• pp.8893-8900
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• 2014

Dysregulated expression of microRNAs (miRNAs) has been shown to be closely associated with tumor development, progression, and carcinogenesis. However, their clinical implications for gastric cancer remain elusive. To investigate the hypothesis that genome-wide alternations of miRNAs differentiate gastric cancer tissues from those matched adjacent non-tumor tissues (ANTTs), miRNA arrays were employed to examine miRNA expression profiles for the 5-pair discovery stage, and the quantitative real-time polymerase chain reaction (qRTPCR) was applied to validate candidate miRNAs for 48-pair validation stage. Furthermore, the relationship between altered miRNA and clinicopathological features and prognosis of gastric cancer was explored. Among a total of 1,146 miRNAs analyzed, 16 miRNAs were found to be significantly different expressed in tissues from gastric cancer compared to ANTTs (p<0.05). qRT-PCR further confirmed the variation in expression of miR-193b and miR-196a in the validation stage. Down-expression of miR-193b was significantly correlated with Lauren type, differentiation, UICC stage, invasion, and metastasis of gastric cancer (p<0.05), while over-expression of miR-196a was significantly associated with poor differentiation (p=0.022). Moreover, binary logistic regression analysis demonstrated that the UICC stage was a significant risk factor for down-expression of miR-193b (adjusted OR=8.69; 95%CI=1.06-56.91; p=0.043). Additionally, Kaplan-Meier survival curves indicated that patients with a high fold-change of down-regulated miR-193b had a significantly shorter survival time (n=19; median survival=29 months) compared to patients with a low fold-change of down-regulated miR-193b (n=29; median survival=54 months) (p=0.001). Overall survival time of patients with a low fold-change of up-regulated miR-196a (n=27; median survival=52 months) was significantly longer than that of patients with a high fold-change of up-regulated miR-196a (n=21; median survival=46 months) (p=0.003). Hence, miR-193b and miR-196a may be applied as novel and promising prognostic markers in gastric cancer.

• Asian-Australasian Journal of Animal Sciences
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• 제31권4호
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• pp.595-606
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• 2018

Objective: The Fusarium mycotoxins of deoxynivalenol (DON) and zerolenone (ZEN) cause health hazards for both humans and farm animals. Therefore, the main intention of this study was to reveal DON and ZEN effects on the mRNA expression of pro-inflammatory cytokines and other immune related genes in the liver of piglets. Methods: In the present study, 15 six-week-old piglets were randomly assigned to the following three different dietary treatments for 4 weeks: control diet, diet containing 8 mg DON/kg feed, and diet containing 0.8 mg ZEN/kg feed. After 4 weeks, liver samples were collected and sequenced using RNA-Seq to investigate the effects of the mycotoxins on genes and gene networks associated with the immune systems of the piglets. Results: Our analysis identified a total of 249 differentially expressed genes (DEGs), which included 99 upregulated and 150 downregulated genes in both the DON and ZEN dietary treatment groups. After biological pathway analysis, the DEGs were determined to be significantly enriched in gene ontology terms associated with many biological pathways, including immune response and cellular and metabolic processes. Consistent with inflammatory stimulation due to the mycotoxin-contaminated diet, the following Kyoto encyclopedia of genes and genomes pathways, which were related to disease and immune responses, were found to be enriched in the DEGs: allograft rejection pathway, cell adhesion molecules, graft-versus-host disease, autoimmune thyroid disease (AITD), type I diabetes mellitus, human T-cell leukemia lymphoma virus infection, and viral carcinogenesis. Genome-wide expression analysis revealed that DON and ZEN treatments downregulated the expression of the majority of the DEGs that were associated with inflammatory cytokines (interleukin 10 receptor, beta, chemokine [C-X-C motif] ligand 9), proliferation (insulin-like growth factor 1, major facilitator superfamily domain containing 2A, insulin-like growth factor binding protein 2, lipase G, and salt inducible kinase 1), and other immune response networks (paired immunoglobulin-like type 2 receptor beta, Src-like-adaptor-1 [SLA1], SLA3, SLA5, SLA7, claudin 4, nicotinamide N-methyltransferase, thyrotropin-releasing hormone degrading enzyme, ubiquitin D, histone $H_2B$ type 1, and serum amyloid A). Conclusion: In summary, our results demonstrated that high concentrations DON and ZEN disrupt immune-related processes in the liver.

• 한국미생물학회:학술대회논문집
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• 한국미생물학회 2002년도 추계학술대회
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• pp.162-172
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• 2002

Schizosaccharomyces pombe is suited for the study of cytokinesis as it divides by forming a septum in the middle of the cell at the end of mitosis. To enhance our understanding of the cytokinesis, we have carried out a genetic screen for temperature-sensitive S. pombe mutants that show defects in septum formation and cell division. Here we present the isolation and characterization of a new temperature-sensitive mutant, sun1(septum uncontrolled), which undergoes uncontrolled septation during cell division cycle at restrictive temperature $(37^{\circ}C)$. In sun1 mutant, actin ring and septum are positioned at random locations and angles, and nuclear division cycle continues. These observations suggest that the sun] gene product is required for the proper placement of the actin ring as well as precise septation. The sun] mutant is monogenic recessive mutation unlinked to previously known various cdc genes of S. pombe. In a screen for $sunl^+$ gene to complement the sun] mutant, we have cloned a gene, $susl^+$(suppressor of sun1 mutant), that encodes a protein of 689 amino acids. The predicted amino acid sequence of $susl^+$ gene is similar to the human hMadlp and Saccharomyces cerevisiae Mad1p, a component of the spindle checkpoint in eukaryotic cells. The null mutant of $susl^+$ gene grows normally at various temperatures and has the increased sensitivity to anti-microtubule drug, while $susl^+$ mutant shows no sensitivity to microtubule destabilizing drugs. The putative S. pombe Sus1p directly interacts with S. pombe Mad2p in yeast two-hybrid assays. These data suggest that the newly isolated susr gene encodes S. pombe Mad1p and suppresses sun] mutant defective in controlled septation in a cell division cycle.

• Journal of Animal Science and Technology
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• 제51권5호
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• pp.413-420
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• 2009

본 연구는 성체우의 FS, MS, C-MS를 배양액에 첨가하여 세포증식과 난포의 성장과 배란에 미치는 영향을 관찰하기 위하여 실시하였다. 세포증식은 세포수와 MTT assay을 실시하여 확인하였다. 그 결과 세포증식은 혈청첨가 배양액에 따라 유의적인 차이를 나타내었다. 특히 근육위성세포의 세포증식은 MS를 첨가한 배양액에서 높은 반면 면역세포의 증식은 FBS에서 배양한 세포에 비해 낮은 것을 확인하였다. 또한 난포의 성장 및 배란을 관찰한 결과 난포의 성장에 유의한 차이가 없었으나 군간 차이를 보였으며 배란율과 비례적이었고, 배란율은 FBS와 C-MS가 첨가된 배양액에서 유의적으로 높은 것을 관찰할 수 있었다(P<0.05). FBS군과 C-MS 군 간에는 차이가 없었다. 3T3-L1 세포에서 창상치유는 FS에서 배양한 세포에서 빠른 회복을 나타냈으며, 증식은 MS에서 높게 나타났다(p<0.001). 따라서 본 결과는 세포배양 과정에서 세포에 따른 혈청의 선택은 매우 중요하다는 근거자료를 제시 하였으며, 분리 된 성별 특이 한우 혈청은 FBS의 대체 물질로서 사용이 가능할 것으로 사료된다.

• 대한바이러스학회지
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• 제28권3호
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• pp.247-265
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• 1998

To determine the sequence and expression of the VP7 gene of Korean isolates (CAU-9), viral RNA was purified and used for cDNA amplification by RT-PCR. The VP7 cDNA was cloned, sequenced, and expressed using baculovirus expression system. The result showed that the sequence homologies CAU-9 compared with foreign isolated strains Wa, 417, TMC-II, 95B and SA11 were ranged from 74.0% to 95.1 % of nucleotide sequence and 35% to 43% of amino acid sequence, respectively. High homology of CAU-9 was observed in Japanease isolates 417 (nucleotide sequence homology was 95.1% and amino acid sequence homology was 43%). To express VP7 gene, the VP7 cDNA was cloned into pCR-Bac vector and inserted into the genome of baculovirus adjacent to the polyhedrin promoter by cotransfection of Spodoptera frugiperda (Sf9) insect cells with wild type baculovirus DNA. In antigenic analysis of Sf9 cells inoculated with the recombinant VP7, immunofluorescence assay revealed positive for viral antigens. In metabolic labeling of Sf9 cell lysates infected with recombinant baculoviruses, it was revealed that the protein of 34 kDa was expressed. The limited study of expressed VP7 protein inoculated with guinea pigs failed to elicit neutalizing antibody. As a results, the sequence analysis and expression of VP7 protein of rotavirus CAU-9 isolated from an infant in Korea could permit the conformation and development of virus like particles which may be useful in designing vaccine strategy.

• Journal of Plant Biotechnology
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• 제34권4호
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• pp.347-354
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• 2007

유전자변형 작물의 상업화를 위해서는 유전자변형 작물이 식품으로서의 안전성과 환경에 미치는 영향에 관한 평가가 이루어져야한다. 이를 위해 개발자는 유전자변형 작물의 방출이전에 유전자변형 작물 개발에 관한 정보를 제출해야만 한다. 본 연구는 유전자변형 작물의 환경 위해성 평가를 위한 분자생물학적 자료를 제공하고자 수행하였다. 아그로 박테리움 형질전환법을 통하여 해충저항성 CryIAc 유전자가 도입된 배추 형질전환체를 획득한 후, 분자생물학적인 분석을 통하여 유전자의 copy수, 안정성, 식물체내에서의 발현을 확인하였고, T-DNA의 배추게놈내의 인접서열을 분석하였다. T-DNA의 게놈내 삽입 인접서열을 바탕으로 유전자변형 배추를 동정할 수 있는 프라이머를 제작하였고, 이를 이용한 검정방법을 수립하였다. 계통 특이 프라이머를 이용한 해충저항성 배추 후대의 PCR 분석결과와 제초체 처리결과가 서로 일치하였다. 본 연구 결과로 환경위해성 평가를 위한 해충저항성 배추의 분자생물학적인 자료를 획득하였으며, 개발된 프라이머는 해충저항성 배추의 검출을 위하여 유용하게 사용할 수 있음을 확인하였다.

• 대한바이러스학회지
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• 제27권1호
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• pp.19-27
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• 1997

본 연구에서는 영장류에서 감염성 면역결핍 증상을 일으키는 type D simian retrovirus (SRV)를 검출하기 위해 SRV env 유전자의 특정 지역을 선정하여 증폭, 검색하는 중합효소 연쇄반응 (PCR)법을 개발하였다. 증폭반응의 대상부위인 SRV env 유전자의 3' 후반부는 서로 다른 3 종류의 SRV subtype 1, 2, 그리고 subtype 3에 걸쳐 높은 보존율을 보이고 있다. 반응 결과, 1차 PCR 만으로 3 종류의 SRV subtypes를 동시에 검출, 증폭하였으며 SRV와 더불어 영장류에서 감염성 면역결핍을 유도하는 주요 바이러스 simian immunodeficiency virus 또는 simian T-Iymphotropic virus type 1에 감염된 영장류의 peripheral blood mononuclear cells (PBMCs)을 비교, 완전한 증폭 특이성이 확인되었고 교차반응으로 인한 위양성반응은 발견되지 않았다. 한편, 위 증폭반응의 검출 민감도 측정을 위해 준정량적 적정 PCR을 수행하였으며 SRV 게놈과 증폭 대상 부위의 분자량을 기준으로 한 본 PCR법의 검출 한계는 단 한번의 증폭과 간단한 ethidium bromide 염색만으로 적어도 $5-7{\times}10^4$ 개의 PBMCs 중 한 개의 감염세포를 검출할 수 있는 것으로 확인되었다. 본 연구에서 확인된 신속성과 반응 특이성, 그리고 높은 민감도의 본 PCR 법은 현재 유일한 AIDS 연구모델인 영장류의 SIV 감염 연구 전후에 반드시 필요한 효과적인 SRV 감염검색에서 ELISA법의 위양성에 의한 약점을 보완하고 보다 높은 검출 민감도를 확보함으로써 연구모델 실험의 오류를 최소화하는 데 중요한 역할을 하게 될 것이다.

• Molecular & Cellular Toxicology
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• 제3권2호
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• pp.98-106
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• 2007

Genotoxic stress triggers a variety of biological responses including the transcriptional activation of genes regulating DNA repair, cell survival and cell death. In this study, we investigated to examine gene expression profiles and genotoxic response in TK6 cells treated with DNA damaging agents MNNG (N-methyl-N'-nitrosoguanidine) and hydrogen peroxide $(H_2O_2)$. We extracted total RNA in three independent experiments and hybridized cRNA probes with oligo DNA chip (Applied Biosystems Human Genome Survey Microarray). We analyzed raw signal data with R program and AVADIS software and identified a number of deregulated genes with more than 1.5 log-scale fold change and statistical significancy. We indentified 14 genes including G protein alpha 12 showing deregulation by MNNG. The deregulated genes by MNNG represent the biological pathway regarding MAP kinase signaling pathway. Hydrogen peroxide altered 188 genes including sulfiredoxins. These results show that MNNG and $H_2O_2$ have both uniquely regulated genes that provide the potential to serve as biomarkers of exposure to DNA damaging agents.