• 생명과학회지
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• 제19권3호
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• pp.362-365
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• 2009

Hepatocyte nuclear factor-$4{\alpha}$ (HNF-$4{\alpha}$)는 췌장 베타세포의 발생, 분화에 중요한 역할을 하는 전사인자로서, 그 유전자 변이와 MODY와의 연관성이 보고된 바 있다. 특히, 일본인의 경우에는 HNF-$4{\alpha}$ 12352 C>T가 제2형 당뇨병과의 연관성이 있다고 보고되어 있다. 이에 본 연구에서는 HNF-$4{\alpha}$의 유전자 변이와 제2형 당뇨병 사이에 어떠한 연관성이 한국인에서도 있는지 알아보았다. 그 결과, HNF-$4{\alpha}$의 유전자 변이인 12352 C>T와 제2형 당뇨병사이에는 구체적인 연관성이 없는 것으로 확인되었다. 이는 제2형 당뇨병과의 연관성이 있다고 보고된 이전의 일본인이나 유럽인을 대상으로 한 연구결과와 다소 차이가 있는 것이며, 한국인을 위한 고유의 통계적 data로서 의미가 있는 것이라 하겠다. 이러한 차이를 보이는 원인을 유추해본다면, 인종에 따라 서로 다른 유전적 그리고 환경적인 요인의 차이에 의해 다형성이 미치는 영향력에 차이가 있기 때문으로 생각된다. 이는 당뇨병과 같은 질환이 특정한 한 두 개의 유전자 이상에 의해서 발생되기 보다는, 여러 유전자들이 상호 작용함으로써 발생할 뿐만 아니라 환경적인 요인의 영향을 많이 받기 때문에 인종이나 민독과 같은 인구 집단 사이에 유전인자의 차이가 생기는 것으로도 판단된다. 따라서 연구결과의 임상적인 의의를 확실하게 확인하기 위하여서는 향후 대상 환자 수와 대조군을 더욱 늘리고 다양한 HNF-$4{\alpha}$ 유전자 다형성을 후보로 하여 추가적인 연구를 할 필요가 있을 것으로 생각된다.

• Childhood Kidney Diseases
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• 제15권1호
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• pp.38-48
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• 2011

목 적: 국소성 분절성 사구체 경화증(Focal segmental glomerulosclerosis, 이하 FSGS)은 소아신부전의 원인 중 가장 흔한 사구체 질환이다. 일차성 FSGS의 병인은 아직 알려져 있지 않으므로, 저자들은 FSGS의 동물 모델을 대상으로 cDNA 마이크로어레이를 이용한 유전자 발현 양상 분석을 통하여 유전자 발현 수준에서의 FSGS의 질환의 특성을 밝히고자 하였다. 방 법: 사람의 일차성 FSGS와 유사한 질병경과를 보이는 동물모델인 FGS/kist 생쥐의 신피질 조직을 대조군 생쥐(FGS/kist 생쥐의 조상 strain인 RFM/kist 생쥐)와 AB 1700 mouse chip을 이용한 마이크로어레이 실험으로 비교하였다. 결 과: FGS 질병특이 유전자가 62개 추출되었다. 이들은 세포주기/사멸, 면역반응과 지질 대사/혈관 질환과 관련된 유전자들로써, 유전자간 network의 중심유전자가 면역반응(TNF, IL-6/4, IFNg)과 세포사멸 조절 유전자(TP 53), 그리고 지질대사의 중요 유전자인 PPARG이었다. 결 론: 이 연구에서 저자들은 자발적인 FSGS의 임상경과를 보이는 FGS/Kist 생쥐의 신장조직의 유전자 발현의 분석을 통하여 신장세포사멸과 면역반응에 뒤따르는 기질 섬유화, 그리고 지질 대사의 이상과 조기 혈관 질환이 FSGS의 병태생리에 기여할 것임을 다시 확인할 수 있었다. 추가적인 연구가 계속된다면 global transcriptome profiling 기법으로 병인 탐색 및 치료방법 개발 에 의미 있는 결과를 도출할 수 있을 것이다.

• 동의신경정신과학회지
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• 제18권1호
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• pp.95-110
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• 2007

Objective : The purpose of this study was to investigate molecular basis of neuroprotective effect in total ginsenosides. After H202 induced neurotoxicity, gene expression profiling of SH-SY5Y neuroblastoma cells treated by total ginsenosides is analyzed. Method : After SH-SY5Y cells were cultured, they were damaged by H202 induced oxidative stress. After twenty four hours, experimental group is treated by total ginsenosides and control group is treated by 0.9% saline. A high density cDNA microarray chip is used to analyze the gene expression profiling of SH-SY5Y cells. The Significance Analysis of Microarray method is used for identifying genes on a microarray. Results : 1. According to the results of microarray experiment, 17 genes were up-regulated, 38 genes were down-regulated. 2. Expression of OPHNl, KTANl, ATM, PRKCE, MAPKs genes associated with cell proliferation, neural growth, and the prevention of apoptosis were increased. 3. Change of EPX gene was the greatest among all genes. EPX gene associated with oxidative stress, and tumor suppressor gene ADAM11 were decreased. Conclusion : According to this study, molecular basis of neuroprotective effect of total ginsenosides is as followings: the increase of gene expression associated with cell proliferation, neuron growth, the prevention of apoptotsis and decrease of gene expression associated with oxidative stress and tumor suppressor.

• Reproductive and Developmental Biology
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• 제37권1호
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• pp.1-8
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• 2013

In order to investigate genetic stability and gene expression profile after cloning procedure, two groups of cloned pigs were used for swine leukocyte antigen (SLA) gene nucleotide alteration and microarray analyses. Each group was consist of cloned pigs derived from same cell line (n=3 and 4, respectively). Six SLA loci were analyzed for cDNA sequences and protein translations. In total, 16 SLA alleles were identified and there were no evidence of SLA nucleotide alteration. All SLA sequences and protein translations were identical among the each pig in the same group. On the other hand, microarray assay was performed for profiling gene expression of the cloned pigs. In total, 43,603 genes were analyzed and 2,150~4,300 reliably hybridized spots on the each chip were selected for further analysis. Even though the cloned pigs in the same group had identical genetic background, 18.6~47.3% of analyzed genes were differentially expressed in between each cloned pigs. Furthermore, on gene clustering analysis, some cloned pigs showed abnormal physiological phenotypes such as inflammation, cancer or cardiomyopathy. We assumed that individual environmental adaption, sociality and rank in the pen might have induced these different phenotypes. In conclusion, the results of the present study indicate that SLA locus genes appear to be stable following SCNT. However, gene expressions and phenotypes between cloned pigs derived from the same cell line were not identical even under the same rearing conditions.

• Clinical and Experimental Reproductive Medicine
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• 제39권2호
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• pp.87-93
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• 2012

Objective: Lin28 has been known to control the proliferation and pluripotency of embryonic stem cells. The purpose of this study was to determine the downstream effectors of Lin28 in mouse embryonic stem cells (mESCs) by RNA interference and microarray analysis. Methods: The control siRNA and Lin28 siRNA (Dharmacon) were transfected into mESCs. Total RNA was prepared from each type of transfected mESC and subjected to reverse transcription-polymerase chain reaction (RT-PCR) analysis to confirm the downregulation of Lin28. The RNAs were labeled and hybridized with an Affymetrix Gene-Chip Mouse Genome 430 2.0 array. The data analysis was accomplished by GenPlex 3.0 software. The expression levels of selected genes were confirmed by quantitative real-time RT-PCR. Results: According to the statistical analysis of the cDNA microarray, a total of 500 genes were altered in Lin28-downregulated mESCs (up-regulated, 384; down-regulated, 116). After differentially expressed gene filtering, 31 genes were selected as candidate genes regulated by Lin28 downregulation. Among them, neuropeptide Y5 receptor and oocyte-specific homeobox 5 genes were significantly upregulated in Lin28-downregulated mESCs. We also showed that the families of neuropeptide Y receptor (Npyr) and oocyte-specific homeobox (Obox) genes were upregulated by downregulation of Lin28. Conclusion: Based on the results of this study, we suggest that Lin28 controls the characteristics of mESCs through the regulation of effectors such as the Npyr and Obox families.

• Parasites, Hosts and Diseases
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• 제49권4호
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• pp.341-347
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• 2011

Acanthamoeba infection is difficult to treat because of the resistance property of Acanthamoeba cyst against the host immune system, diverse antibiotics, and therapeutic agents. To identify encystation mediating factors of Acanthamoeba, we compared the transcription profile between cysts and trophozoites using microarray analysis. The DNA chip was composed of 12,544 genes based on expressed sequence tag (EST) from an Acanthamoeba ESTs database (DB) constructed in our laboratory, genetic information of Acanthamoeba from TBest DB, and all of Acanthamoeba related genes registered in the NCBI. Microarray analysis indicated that 701 genes showed higher expression than 2 folds in cysts than in trophozoites, and 859 genes were less expressed in cysts than in trophozoites. The results of real-time PCR analysis of randomly selected 9 genes of which expression was increased during cyst formation were coincided well with the microarray results. Eukaryotic orthologous groups (KOG) analysis showed an increment in T article (signal transduction mechanisms) and O article (posttranslational modification, protein turnover, and chaperones) whereas significant decrement of C article (energy production and conversion) during cyst formation. Especially, cystein proteinases showed high expression changes (282 folds) with significant increases in real-time PCR, suggesting a pivotal role of this proteinase in the cyst formation of Acanthamoeba. The present study provides important clues for the identification and characterization of encystation mediating factors of Acanthamoeba.

• 대한전기학회:학술대회논문집
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• 대한전기학회 2006년도 제37회 하계학술대회 논문집 C
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• pp.1645-1646
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• 2006

Micro ElectroMechanical Systems (MEMS) 기술을 이용한 초미세 유체 제어 시스템 (마이크로 펌프, 마이크로 밸브, 마이크로 채널, 마이크로 믹서 등)은 화학, 생명분야의 DNA 분석, 항원-항체 분석, 질병의 진단 등에 사용되는 lab-on-a-chip, micro total analysis system ($\mu$-TAS) 등에서 화학 및 바이오 유체를 제어하는 분석 시스템의 일부분으로서 사용되며 필수적으로 요구된다. 본 논문에서는 이러한 microchip을 구현하기 위해 초미세 유체 제어 소자인 마이크로 펌프와 밸브를 같은 기관 위에 polydimethylsiloxane (PDMS)와 indium tin oxide (ITO)-Glass를 사용하여 동일한 구조로 집적 하였다. 마이크로 펌프의 pumping rate은 인가 직류 펄스 전력의 주파수와 duty 비를 변화시켜 최적화하였다. 직류 펄스 전력 500 mW를 인가하였을 때 주파수 2 Hz, duty 비 7 %에서 약 $1.05{\mu}l/min$의 최대 유량이 측정되었다. 마이크로 밸브는 ITO 히터에 전력을 인가함으로서 유량의 on/off 제어가 잘 됨을 확인할 수 있었고 유체를 closing하기 위해 필요한 전력은 약 300 mW이다.

• Clinical and Experimental Pediatrics
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• 제48권2호
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• pp.197-203
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• 2005

목 적 : MMP 활동의 변화는 류마치스 관절염과 암 전위를 비롯한 여러 질환에서 보고되고 있다. 최근에 확장성 심근증에서 MMP 활동의 증가가 발표되었다. 아드리아마이신으로 유도된 심근증에서 MMP에 대한 보고는 없는 실정이다. 아드리아마이신으로 유도된 심근증에서 MMP, TIMP 유전자 발현을 연구하고 cytokine과의 관련성을 알아보고자 본 연구를 실시하였다. 방 법 : Sprague Dawley 쥐에 아드리아마이신 5 mg/kg을 1주일에 2번씩 2주간(누적 용량 : 20 mg/kg) 복강내 주사하였고, 정상쥐를 대조군으로 하였다. 2주 후에 쥐를 희생시켜서 혈청과 심장조직을 얻었다. 혈청에서 ELISA 원리로 MMP-2, TIMP-3, IL-6, TNF-${\alpha}$를 측정하였다. 심장에서 total RNA를 추출하였고, MMP-2, TIMP-3 IL-6, TNF-${\alpha}$ primer를 이용하여 PCR로 증폭하였다. 증폭된 DNA는 1% agarose gel에서 전기 영동하였고 UV light 아래에서 필름으로 촬영하였다. 결 과 : 혈청 MMP-2와 TIMP-3는 두 군 간에 유의한 차이가 없었다. 아드리아마이신군에서 IL-6은 $36.8{\pm}2.8pg/mL$, TNF-${\alpha}$은 $2.2{\pm}2.7pg/mL$로 정상군에 비해 유의한 증가를 보였다. 혈청 MMP-2와 TNF-${\alpha}$와는 r=0.41로 유의한 상관관계가 있었다. 심근 조직에서 MMP-2, IL-6, TNF-${\alpha}$는 발현되지 않았고, TIMP-3는 아드리아마이신군에서 대조군에 비해 유전자 발현이 감소되었다. 결 론 : 아드리아마이신으로 유도된 급성 심근증 모델에서는 심근에서 MMP, IL-6, TNF-${\alpha}$가 발현되지 않았고, TIMP 발현이 감소함을 알 수 있었다. 혈청 MMP와 TNF-${\alpha}$와의 상관성이 유의하게 높았으므로 TNF-${\alpha}$가 MMP 발현을 조절함을 시사해 준다. 앞으로 만성 심근증 모델에서 MMP, TIMP 발현에 대하여 연구할 예정이다.

• Korean Journal of Acupuncture
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• 제23권3호
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• pp.133-160
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• 2006

Objectives : Scolopendrae Corpus has a broad array of clinical applications in Korean medicine, including treatment of inflammatory conditions such as arthritis. To explore the global gene expression profiles in human Raw cell lines treated with Scolopendrae Corpus herbal-acupuncture solution (SCHAS), cDNA microarray analysis was performed. Methods : The Raw 264.7 cells were treated with lipopolysaccharide (LPS), SCHAS, or both. The primary data was normalized by the total spots of intensity between two groups, and then normalized by the intensity ratio of reference genes such as housekeeping genes in both groups. The expression ratio was converted to log2 ratio. Normalized spot intensities were calculated into gene expression ratios between the control and treatment groups. Greater than 2 fold changes between two groups were considered to be of significance. Results : Of the 8 K genes profiled in this study, with a cut-off level of two-fold change in the expression, 20 genes (BCL2-related protein A1, MARCKS-like 1, etc.) were upregulated and 5 genes (activated RNA polymerase II transcription cofactor 4, calcium binding atopy-related autoantigen 1, etc.) downregulated following LPS treatment. 139 genes (kell blood group precursor (McLeod phenotype), ribosomal protein S7, etc.) were upregulated and 42 genes (anterior gradient 2 homolog (xenopus laevis), phosphodiesterase 8B, etc.) were downregulated following SCHAS treatment. And 10 genes (yeast saccharomyces cerevisiae intergeneic sequence 4-1, mitogen-activated protein kinase 1, etc.) were upregulated and 8 genes (spermatid perinuclear RNA binding protein, nuclear receptor binding protein 2, etc.) were downregulated following co-stimulation of SCHAS and LPS. Discussions : It is thought that microarrays will play an ever-growing role in the advance of our understanding of the pharmacological actions of SCHAS in the treatment of arthritis. But further studies are required to concretely prove the effectiveness of SCHAS.

• IMMUNE NETWORK
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• 제5권4호
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• pp.205-214
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• 2005

Background: We examined global gene expression profiles of peripheral blood mononuclear cells (PBMCs) in patients with ulcerative colitis (DC), and tested whether the identified genes with the altered expression might be associated with susceptibility to UC. Methods: PBMCs from 8 UC and 8 normal healthy (NH) volunteers were collected, and total RNAs were subjected to the human 8.0K cDNA chip for the micro array analysis. Real time-PCR (RT-PCR) was performed to verify the results of micro array. One hundred forty UC patients and 300 NH controls were recruited for single nucleotide polymorphism (SNP) analysis. Results: Twenty-five immune function-related genes with over 2-fold expression were identified. Of these genes, two chemokines, namely, CXCL1 and CCL20, were selected because of their potential importance in the evocation of host innate and adaptive immunity. Four SNPs were identified in the promoter and coding regions of CXCL1, while there was no significant difference between all patients with UC and controls in their polymorphisms, except minor association at g.57A>G (rs2071425, p=0.02). On the other hand, among three novel and one known SNPs identified in the promoter region of CCL20, g. -1,706 G>A (p=0.000000055), g. -1,458 G>A (p=0.0048), and g. -962C>A (p=0.0006) were found to be significantly associated with the susceptibility of Uc. Conclusion: Altered gene expression in mononuclear cells may contribute to IBD pathogenesis. Although the findings need to be confirmed in other populations with larger numbers of patients, the current results demonstrated that polymorphisms in the promoter region of CCL20 are positively associated with the development of Uc.