경간동맥화학색전술로 치료받은 간암환자의 생존율과 관련된 예후인자 분석

임정묵,이태원,박선미,윤세진,박길선,채희복,이헌영 충남대학교 의과대학 의학연구소 2002 충남의대잡지 Vol.29 No.1

Transarterial Chemoembilization (TACE) has been shown to be effective in patients with unresectable hepatocellular carcinoma (HCC) and can be applicable to patients with resectable HCC who reject operation. There have been studies concerning prognostic factors in patients with (HCC) undergoing TACE but they reported diffenrent prognostic factors from each other. The aim of this study is to estimate the survival rates and to determine which prognostic factors contribute to long-term survival after TACE of HCC. Thirty eight patients with HCC who had been treated by TACE were analized retrospectively. TACE was accomplished by hepatic arterial infusion of a suspension of lipiodol and anticancer drugs (Adriamycin), either alone or followed by gelfoam embolization. Male to female ratio was 4.4:l. Mean survival time was 463 days. Maximum survival time was 1,683 days. The overall cumulative survival rates at the end of the first and second years were, respectively, 54% and 38%. According to univariate analysis (log-rand scale test), variables significantly associated with survival were: tumor type, hepatic vein thrombosis, portal vein thrombosis, lipiodol uptake pattern, remained enhancement after TACE, AST, ALT, height and tumor size. Multivariate analysis (Cox proportional hazard model) for the significant variables in a univariate analysis revealed that the AST level, hepatic vein thrombosis, remained enhancement after TACE were statistically significant independent prognostic factors. The criteria of known contraindication may still be applied for TACE treatment of HCC. Additionally we should select the good prognostic patients without hepatic vein thrombosis and try to normalize the AST level if abnormal level before undergoing TACE. Thereafter, intensive chemoembilization to minimize the remained tumor is required to improve survival rates.

S. cerevisiae에서 세포주기의 진행에 관련된 COS28과 COS46 유전자의 클로닝

박정은,임선희,선우양일 동아대학교 기초과학연구소 2000 基礎科學硏究論文集 Vol.17 No.1

본 연구에서는 효모의 돌연변이주를 분리하여 G1/S기에서 세포주기 진행에 관련된 유전자의기작을 이해하기 위해 연구를 수행하였다. 먼저, cos28과 cos46 돌연변이주의 MMS(methylmethane sulfonate)와 HU(hydroxyurea)에 대한 감수성을 조사하였다. 이들 돌연변이주들은 MMS와 HU에 대해서 감수성을 보이므로, S기 checkpoint에 관련되었다고 생각된다. 다음으로 cos28과 cos46 돌연변이를 상보하는 유전자를 클로닝하였고, 서열화하였다. 그리고 클론된 유전자는 데이터 베이스 분석을 통해 조사하였다. cos28 변이주를 상보하는 DNA 단편은 전체 6개의 ORF를 가지며 그 중 기능이 알려진 ORF로는 RPS21B와 LCB3와 MRS3가 있고, 기능이 알려지지 않은 ORF로는 YJL131와 YJL132와 YJL135의 세개를 포함하고 있었다. cos46 유전자는 전체 6개의 ORF를 가지며, 그 중 기능이 알려진 CYP5와 HNT2와 SRB7와 GIC2를 가지고, 기능이 알려지지 않은 ORF로 YDR306, YDR307를 포함하고 있음이 확인되었다. In this study, to understand the mechanisms of genes which related in cell cycle progression at G1/S phase, using the isolated mutants in S. cerevisiae were investigated. First of all, cos28 and cos46 were examined the sensitive to MMS(methylmethane sulfonate) and HU(hydroxyurea). These mutants were showed MMS, HU sensitivities and might be a act at a checkpoint pathway during S phase. These mutants were carried out gene cloning for the gene which were complemented the cos mutants. As a result, the DNA fragments which complement with the cos28 and cos46 mutants, were cloned and sequenced. And then cloned genes were surveyed though data base analysis. Complemented DNA fragment of COS28 contained six ORF which were consisted RPS21B, LCB3, MRS3, YJL131, YJL132, and YJL135. Complemented DNA fragment of COS46 contained six ORF which were consisted CYP5, HNT2, SRB7, GIC2, YDR306 and YDR307.

출아효모에서 TAR 클론닝법을 이용한 고등동물의 게놈으로부터 특정 염색체 부위의 분리

박정은,윤영호,이윤주,김광섭,윤세련,안태진,임선희,선우양일 동아대학교 기초과학연구소 2003 基礎科學硏究論文集 Vol.20 No.1

복잡한 게놈 분석에 용이하도록 효모의 인공 염색체(YAC) 클론닝 시스템은 발달되어왔다. YAC은 박테리아의 인공 염색체(BAC)보다 더 큰 단편을 클론닝할 수 있고, 또한 클론된 단편을 쉽게 변형시킬 수 있다. 형질전환과 연계된 재조합법(Transformation-Associated Recombination; TAR)은 게놈 라이브러리를 만들지 않고 직접 게놈 DNA로부터 분리하고자 하는 유전자나 특정 염색체 부분을 클론닝 할 수 있는 방법이다. 이 방법은 spheroplast transformation을 수행하는 동안, 목적으로 하는 유전자의 5' 그리고 3' 염기 배열(hooks)을 지닌 TAR 벡터와 게놈 DNA 사이에서 일어나는 상동성 재조합에 의해 이루어진다. 효모 내의 in vivo 재조합을 이용한 TAR 클론닝법은 복잡한 게놈으로부터 목적의 염색체 부분을 원형 YAC의 형태로서 선택적으로 분리할 수 있다. 그러므로 TAR 클론닝 법은 특정 염색체로부터 YACs을 만드는데 매우 유용하여, 전체 게놈으로부터 특이적 유전자나 유전자의 family를 분리하는데 효과적인 방법으로 사료된다. Yeast artificial chromosome (YAC) cloning systems have advanced the analysis of complex genomes considerably. They permit the cloning of larger fragments than do bacterial artificial chromosome systems, and the cloned material is more easily modified. Transformation-associated recombination (TAR) is a cloning technique that allows specific chromosomal regions or genes to be isolated directly from genomic DNA without prior construction of a genomic library. This technique involves homologous recombination during spheroplast transformation between genomic DNA and a TAR vector that has 5' and 3' gene targeting sequences (hooks). Using in vivo recombination in yeast, TAR cloning selectively isolates, as circular YACs, desired chromosome segments or entire genes from complex genomes. We propose that TAR cloning can provide an efficient means for generating YACs from specific chromosomes and that TAR cloning may be useful for isolating families of genes and specific genes from total genome DNA.

Functional Classification of Gene Expression Profiles During Differentiation of Mouse Embryonic Cells on Monolayer Culture

Leem, Sun-Hee,Ahn, Eun-Kyung,Heo, Jeong-Hoon The Korean Society for Integrative Biology 2009 Animal cells and systems Vol.13 No.2

Embryonic stem (ES) cells have a capability to generate all types of cells. However, the mechanism by which ES cells differentiate into specific cell is still unclear. Using microarray technology, the differentiation process in mouse embryonic stem cells was characterized by temporal gene expression changes of mouse ES cells during differentiation in a monolayer culture. A large number of genes were differentially regulated from 1 day to 14 days, and less number of genes were differentially expressed from 14 days to 28 days. The number of up-regulated genes was linearly increased throughout the 28 days of in vitro differentiation, while the number of down-regulated genes reached the plateau from 14 days to 28 days. Most differentially expressed genes were functionally classified into transcriptional regulation, development, extra cellular matrix (ECM),cytoskeleton organization, cytokines, receptors, RNA processing, DNA replication, chromatin assembly, proliferation and apoptosis related genes. While genes encoding ECM proteins were up-regulated, most of the genes related to proliferation, chromatin assembly, DNA replication, RNA processing, and cytoskeleton organization were down-regulated at 14 days. Genes known to be associated with embryo development or transcriptional regulation were differentially expressed mostly after 14 days of differentiation. These results indicate that the altered expression of ECM genes constitute an early event during the spontaneous differentiation, followed by the inhibition of proliferation and lineage specification. Our study might identify useful time-points for applying selective treatments for directed differentiation of mouse ES cells.

Colloquia : THE TRANSCRIPTION OF GENES REQUIRED FOR MEIOTIC RECOMBINATION IN CONTROLLED BY THE SWI6 GENE

Sun Hee Leem,Akio Sugino,Hiroyuki Araki 생화학분자생물학회(구 한국생화학분자생물학회) 1993 생화학분자생물학회 추계학술발표논문집 Vol.- No.-

Genome-wide transcriptome analysis reveals inactivation of Wnt/β-catenin by 3,3′-diindolylmethane inhibiting proliferation of colon cancer cells

LEEM, SUN-HEE,LI, XIU JUAN,PARK, MAN HEE,PARK, BYUNG HYUN,KIM, SOO MI Spandidos Publications 2015 International journal of oncology Vol.47 No.3

A Yeast MRE3/REC114 Gene is Essential for Normal Cell Growth and Meiotic Recombination

Leem, Sun-Hee The Microbiological Society of Korea 1999 The journal of microbiology Vol.37 No.4

We have analyzed the MRE3/REC114 gene of Saccharomyces cerevisiae, previously detected in isolation of mutants defective in meiotic recombination. We cloned the MRE3/REC114 gene by complementation of the meiotic recombination defect and it has been mapped to chormosome XIII. The DNA sequence analysis revealed that the MRE3 gene is identical to the REC114 gene. The upstream region of the MRE3/REC114 gene contains a T_4C site, a URS (upstream repression sequence) and a TR (T-rich) box-like sequence, which reside upstream of many meiotic genes. Coincidentally, northern blot analysis indicated that the three sizes of MRE3/REC114 transcripts, 3.4, 1.4 and 1.2 kb, are induced in meiosis. A less abundant transcript of 1.4 kb is detected in both mitotic and meiotic cells, suggesting that it is needed in mitosis as well as meiosis. To examine the role of the MRE3/REC114 gene, we constructed mre3 disruption mutants. Strains carrying an insertion or null deletion of the MRE3/REC114 gene showed slow growth in nutrient medium and the doubling time of these cells increased approximately by 2-fond compared to the wild-type strain. Moreover, the deletion mutant (${\delta}$mre3) displayed no meiotically induced recombination and no viable spores. The mre3/rec114 spore lethality can be suppressed by spo13, a mutation that causes cells to bypass reductional division. The double-stranded breaks (DSBs) which are involved in initiation of meiotic recombination were not detected in the analysis of meiotic chromosomal DNA from the mre3/rec114 disruptant. From these results we suggest that the MRE3/REC114 gene product is essential in normal growth and in early meiotic stages involved in meiotic recombination.

Phospholipase D Is Not Involved in Rho A-Mediated Activation of Stress Fiber Formation

Leem, Sun-Hee,Shin, In-Cheol,Kweon, Soo-Mi,Kim, Seung-Il,Kim, Jae-Hong,Ha, Kwon-Su Korean Society for Biochemistry and Molecular Biol 1997 Journal of biochemistry and molecular biology Vol.30 No.5

In order to investigate the role of a small GTP-binding protein RhoA in lysophosphatidic acid (LPA)-induced stress fiber formation, C3 ADP-ribosyltransferase was prepared by expressing in E. coli and then applied to Rat-2 fibroblasts. C3 transferase isolated from E. coli was as effective as the toxin from Clostridium botulinum in ADP-ribosylation of RhoA. Incubation of the cells with C3 transferase for 2 days induced ADP-ribosylation of RhoA by a dose-dependent manner, with a sub-maximal induction at $25\;{\mu}g/ml$. As expected, LPA-induced stress fiber formation was completely blocked by pre-incubation with C3 transferase for 2 days. However, exogenously added C3 transferase had no significant effect on the formation of phosphatidylethanol by LPA. These results suggested that phospholipase D was not activated by RhoA in the LPA-induced stress fiber formation.

Analysis of the Novel VNTR Polymorphisms of MUC8 Gene

Ji Sun Lee,Ja Young Kim,Eun Kyung Ahn,Sang Yeop Lee,Yun Hee Jeong,Se Ra Lee,Sun Hee Leem 한국유전학회 2009 Genes & Genomics Vol.31 No.3

Analysis of mucin genes has identified the presence of several features that may represent important functional domains in mucin glycoproteins. In the central region of each mucin, there are a variable number of tandem repeats (VNTR; minisatellites). However, their genomic levels are unclear because of complex genomic properties. We report here the distribution of VNTR and polymorphic analysis of MUC8. We searched for VNTR of MUC8 using the Tandem Repeat Finder program and found nine VNTR motif. Six (MUC8-MS1~MS6) among the nine VNTRs were evaluated in this study. Each VNTR in MUC8 region was analyzed in genomic DNA obtained from 200 unrelated individuals and multi-generational families. All VNTRs (MUC8-MS1, -MS2, -MS3, -MS4, -MS5 and -MS6) were genotyped as polymorphic. The degree of polymorphism within the MUC8-MS5 showed the highest heterozygosity (h=0.786) in the MUC8 region. In order to perform a segregation analysis of the VNTRs in MUC8, we analyzed genomic DNA obtained from two generations of five families and from three generations of two families. Six of the polymorphic VNTRs were transmitted through meiosis following a Mendelian inheritance, which suggests that polymorphic VNTRs could be useful markers for paternity mapping and DNA fingerprinting.

송절(松節) 약침액이 자유기와 금속 이온으로 유도된 인체 저밀도 지단백질의 산화 반응에 미치는 효과

임선희 ( Sun Hee Leem ),이강파 ( Kang Pa Lee ),문진영 ( Jin Young Moon ) 경락경혈학회 2011 Korean Journal of Acupuncture Vol.28 No.2

목적: 이 연구는 관절 및 심혈관계 질환 치료에 사용되는 松節(Pinus densiflora Gnarl)을 약침용 시료로 조제하여 본 약물의 항산화 효능을 규명하고자 하였으며 이를 다양한 시스템에서 검토하였다. 방법: FeCl2-ascorbic acid system에서 흰쥐 간조직의 지질과산화 반응을 관찰하였고, Fenton reaction system에서 자유기에 의한 plasmid DNA 분절을 유도하였다. 또한 deoxyribose assay를 통해 hydroxyl radical 소거능을 관찰하였고, NBT reduction assay로 superoxide radical 소거능을 검토하였다. 또한 human low-density lipoprotein (LDL)의 산화를 유도하기 위해 CuSO4와 AAPH를 사용하였으며 relative electrophoretic mobility (REM) assay로 LDL 산화 억제 효능을 대조 항산화물질과 비교 검토하였다. 결과: 송절 약침액은 자유기에 의한 간조직의 지질과산화(p<0.01)및 DNA 분절을 현저하게 억제하였으며, hydroxyl radical, superoxide radical (p<0.01), nitric oxide 및 peroxynitrite를 강하게 소거하였다. 또한 CuSO4 (IC50=9.2±0.2 μg/mL)와 AAPH (IC50=34.8±5.1 μg/mL)에 의해 유도된 human LDL의 산화를 억제하였고, REM assay에서도 산화 억제 효능을 재확인할 수 있었다. 결론: 송절 약침액은 활성산소종 및 활성질소종를 소거하였고, 지질과산화를 억제하였으며, 특히 human LDL의 산화적 손상을 방어하였다. 이에 본 약물은 자유기에 의한 심혈관의 산화적 손상을 효과적으로 보호할 것으로 판단된다.