Poster Exhibition : Study the Role of CD133 Enriched Cancer Initiating Cells for Radioresistance in Hepatocellular Carcinoma (초)

Lian Shu Piao,Won Hee Hur,Jung Eun Choi,Sung Woo Hong,Si Hyun Bea,Jong Young Choi,Seung Kew Yoon 대한간학회 2009 Clinical and Molecular Hepatology(대한간학회지) Vol.15 No.4(S)

Parkinson’s Disease with Fatigue: Clinical Characteristics and Potential Mechanisms Relevant to α-Synuclein Oligomer

Li-Jun Zuo,Shu-Yang Yu,Fang Wang,Yanghui Xia,Ying-Shan Piao,Yang Du,Teng-Hong Lian,Rui-Dan Wang,Qiu-Jin Yu,Ya-Jie Wang,Xiao-Min Wang,Piu Chan,Sheng-Di Chen,Yongjun Wang,Wei Zhang 대한신경과학회 2016 Journal of Clinical Neurology Vol.12 No.2

Background and Purpose The aim of this study was to identify the clinical characteristics and potential mechanisms relevant to pathological proteins in Parkinson’s disease (PD) patients who experience fatigue. Methods PD patients (n=102) were evaluated using a fatigue severity scale and scales for motor and nonmotor symptoms. The levels of three pathological proteins—α-synuclein oligomer, β-amyloid (Aβ)1-42, and tau—were measured in 102 cerebrospinal fluid (CSF) samples from these PD patients. Linear regression analyses were performed between fatigue score and the CSF levels of the above-listed pathological proteins in PD patients. Results The frequency of fatigue in the PD patients was 62.75%. The fatigue group had worse motor symptoms and anxiety, depression, and autonomic dysfunction. The CSF level of α-synuclein oligomer was higher and that of Aβ1-42 was lower in the fatigue group than in the non-fatigue group. In multiple linear regression analyses, fatigue severity was significantly and positively correlated with the α-synuclein oligomer level in the CSF of PD patients, after adjusting for confounders. Conclusions PD patients experience a high frequency of fatigue. PD patients with fatigue have worse motor and part nonmotor symptoms. Fatigue in PD patients is associated with an increased α-synuclein oligomer level in the CSF

Analysis of Gene Expression in Primary Hepatocellular Carcinoma Using Differentially Displayed Reverse Transcriptase Polymerase Chain Reaction

Lee, Young Chun,Hur, Wonhee,Choi, Jung Eun,Piao, Lian Shu,Hong, Sung Woo,Bae, Si Hyun,Choi, Jong Young,Yoon, Seung Kew The Korean Society of Gastroenterology 2009 대한소화기학회지 Vol.53 No.6

<P>BACKGROUND/AIMS: The investigation of a specific tumor marker for hepatocellular carcinoma (HCC) is needed to examine the carcinogenesis and to select the patients for treatment options. The aim of this study was to find the genes related to HCC. We also examined the expression level of these genes in cancer cell lines and tissue specimens. METHODS: Three pairs of HCC tissue and non-neoplastic hepatic tissue around the HCC were collected from three patients who underwent resection for HCC. Differential display reverse transcriptase-PCR (DD RT-PCR) using GeneFishingTM PCR was used to detect the differences in the gene expression between in HCC tissue and non-neoplatic tissue. Up- or down-regulated genes in HCC tissue were identified through BLAST searches after cloning and sequencing assays. Real-time RT-PCR assay was employed to detect the expression rate in 11 HCC tissues and human cancer cell lines. RESULTS: Differentially expressed 21 genes were identified, and they were classified as genes involved in protein metabolism, ubiquitin-dependent protein catabolism, carbohydrate metabolism, lipid metabolism, DNA repair, and inflammatory response. CONCLUSIONS: We identified differentially expressed genes in HCC, and these genes may play an important role in the study of hepatocarcinogenesis, development of biomarker, and target therapy for HCC.</P>

Functional characterization of human oncoprotein gankyrin in Zebrafish

So Yeon Kim,Wonhee Hur,최정은,Daniel Kim,Jin Sang Wang,Hye-Yeon Yoon,Lian-Shu Piao,윤승규 생화학분자생물학회 2009 Experimental and molecular medicine Vol.41 No.1

Gankyrin is an oncoprotein containing seven ankyrin repeats that is overexpressed in hepatocellular carcinoma(HCC). Gankyrin binds to Mdm2, which results in accelerated ubiquitylation via degradation of p53, and it also plays an important role in cell proliferation. However, little is known about the relationships between p53 levels, cell proliferation, and gankyrin over-expression. In order to investigate the influence of gankyrin protein on p53 and Mdm2 in a zebrafish model, we injected human gankyrin (hgankyrin) containing expression vectors (pCS2-hgankyrin, pCS2- hgankyrin-EGFP) into zebrafish embryos. To measure p53 and Mdm2 expression in hgankyrin-injected embryos, RT-PCR, Northern blot and in-situ hybridization and BrdU immunostaining were used. In addition, to know the effect of hgankyrin on cell proliferation in vitro, cell viability assays such as MTT, trypan blue staining and RT-PCR following transfection of hgankyrin-containing vector into HEK 293 cell line were performed. In vivo results indicated that p53 mRNA levels decreased but those of Mdm2 were not decreased in the presence of hgankyrin. These results suggest that gankyrin downregulates p53 expression and not Mdm2 expression. In the study of cell proliferation, BrdU-positive cells were predominantly increased in the head and tail regions in hgankyrin-injected zebrafish. Additional in vitro studies using trypan blue staining and MTT assay showed that gankyrin-expressing HEK 293 cells proliferated at a faster rate, indicating that gankyrin promotes cell proliferation. Our results demonstrate that hgankyrin overexpression downregulates p53 expression and promotes cell proliferation in zebrafish. Gankyrin may play an important role in tumorigenesis via its effects on p53 and cell proliferation.

Subgenotype and genetic variability in the precore/core regions of hepatitis B virus in Korean patients with chronic liver disease.

Lyoo, Kwang Soo,Hong, Sung Woo,Song, Myeong Jun,Hur, Wonhee,Choi, Jung Eun,Piao, Lian-Shu,Jang, Jeong Won,Bae, Si Hyun,Choi, Jong Young,Park, Jung Wha,Choi, Sang Wook,Yoon, Seung Kew S. Karger 2011 Intervirology Vol.54 No.6

<P>The aim of this study was to determine the prevalence of hepatitis B virus (HBV) subgenotypes, the spectrum of mutations in the precore/core region through phylogenetic analysis, and the relevance of viral characteristics in disease progression in Korean patients.</P>

불멸화된 일차 간세포를 이용한 C형 간염바이러스의 in vitro 배양

최정은 ( Jung Eun Choi ),허원희 ( Won Hee Hur ),신주엽 ( Ju Yeop Shin ),박련숙 ( Lian Shu Piao ),윤승규 ( Seung Kew Yoon ) 대한소화기학회 2008 대한소화기학회지 Vol.52 No.3

Background/Aims: It is essential to develop an in vitro culture model of primary hepatocytes for the study of hepatocellular function and the pathogenesis of hepatitis C virus (HCV) infection. In this study, we have established the immortalized primary human hepatocyte (IPHH) and performed in vitro culture of HCV derived from human patient. Methods: Primary human hepatocytes were isolated from surgically resected liver tissue and then were immortalized by transfection with the SV40 large T antigen. The characterization of the IPHH during culture was analyzed by immunocytochemistry, RT-PCR, Western blot, ELISA, and soft agar assay. Next, sera and/or liver tissue homogenates from surgically resected liver tissues of patients with HCV infection were inoculated for the culture of HCV in IPHH. After HCV RNA extraction from IPHH and culture media, positive or negative stranded HCV RNA was examined by specific nest RT-PCR. Results: IPHH expressed liver-associated proteins but did not express alpha-fetoprotein. Also IPHH showed ammonia removal activity. With regard to its malignant potential, colony formation in soft agar assay was not observed. Next, positive and negative stranded HCV RNAs in IPHH infected with patient`s sera plus liver tissue homogenates were clearly detected whereas those in IPHH infected with only patient`s sera were not detected. Conclusions: These results demonstrated the phenotypic characteristics of IPHH and the feasibility in vitro culture system of HCV infected human samples. This system might be useful for study of pathogenesis of HCV infection or hepatocyte-based applications. (Korean J Gastroenterol 2008;52:150-160)

차별 발현 역전사중합효소연쇄반응법을 이용한 원발 간세포암종의 유전자 발현 분석

이영춘 ( Young Chun Lee ),허원희 ( Won Hee Hur ),최정은 ( Jung Eun Choi ),박련숙 ( Lian Shu Piao ),홍성우 ( Sung Woo Hong ),배시현 ( Si Hyun Bae ),최종영 ( Jong Young Choi ),윤승규 ( Seung Kew Yoon ) 대한소화기학회 2009 대한소화기학회지 Vol.53 No.6

목적: 원발 간세포암종에서 특이하게 과발현되는 특정유전자를 규명하는 것은 간세포암종 발암 기전의 연구나, 암의 조기진단 및 암 특이 유전자를 표적으로 한 유전자 치료에 매우 중요한 정보를 제공한다. 이에 이번 연구에서는 차별 발현 역전사중합효소연쇄반응법을 활용한 유전자 분석을 통해 간세포암에서 특이하게 과발현되는 특이 유전자를 밝혀내고 이 유전자의 발현율을 여러 시료에서 확인하여 간세포암 관련 유전자들의 기능을 탐색하고자 하였다. 대상 및 방법: 원발 간세포암종으로 수술을 시행 받았던 3명의 간세포암 환자의 간 적출물에서 간세포암종 조직과 암주변의 비 간세포암 조직 3쌍을 채취한 후 차별 발현 역전사중합효소연쇄반응법을 이용하여 간세포암종 조직에서 과발현되거나 소실된 유전자 혹은 저발현되어 있는 유전자를 겔로부터 DNA를 용출하여 이를 클로닝한 후 염기서열분석을 통하여 나타난 유전자의 서열을 유전자 검색프로그램에 넣어 그 유전자의 이름이나 특성들을 탐색하였다. 이후 새로 규명된 유전자의 기능을 검증하기 위하여 11쌍의 간세포암종 및 비간세포암종 조직과 인간 암세포주인 간세포암종 세포주[HepG2, Hep3B, Huh7], 자궁암 세포주[HeLa], 대장암 세포주[HT1299], 섬유육종 세포주[HT1080], 폐암 세포주[A549]에서 실시간 중합효소연쇄반응을 이용하여 이들 유전자의 발현을 확인한 후 분석하였다. 결과: 이번 연구를 통해서 단백 대사, 유비퀴틴 의존성 단백 대사, 탄수화물 대사, 지방대사, 유전자복구, 염증 반응 등의 다양한 기능을 가지는 21종의 유전자들을 탐색하였다. 결론: 이번 연구를 통하여 간세포암종 조직과 비간세포암종 조직에서 발현차이를 보이는 유전자를 탐색하였고, 이러한 유전자의 특성 규명은 간세포암종의 발암 기전을 규명하거나 조기 간세포암을 진단하는 바이오 마커의 개발 혹은 분자표적을 대상으로 하는 유전자 치료의 개발에 기초자료로 유용할 것으로 생각한다. Background/Aims: The investigation of a specific tumor marker for hepatocellular carcinoma (HCC) is needed to examine the carcinogenesis and to select the patients for treatment options. The aim of this study was to find the genes related to HCC. We also examined the expression level of these genes in cancer cell lines and tissue specimens. Methods: Three pairs of HCC tissue and non-neoplastic hepatic tissue around the HCC were collected from three patients who underwent resection for HCC. Differential display reverse transcriptase-PCR (DD RT-PCR) using GeneFishing(TM) PCR was used to detect the differences in the gene expression between in HCC tissue and non-neoplatic tissue. Up- or down-regulated genes in HCC tissue were identified through BLAST searches after cloning and sequencing assays. Real-time RT-PCR assay was employed to detect the expression rate in 11 HCC tissues and human cancer cell lines. Results: Differentially expressed 21 genes were identified, and they were classified as genes involved in protein metabolism, ubiquitin-dependent protein catabolism, carbohydrate metabolism, lipid metabolism, DNA repair, and inflammatory response. Conclusions: We identified differentially expressed genes in HCC, and these genes may play an important role in the study of hepatocarcino-genesis, development of biomarker, and target therapy for HCC. (Korean J Gastroenterol 2009;53:361-368)