Influence of Reduced Folate Carrier and Dihydrofolate Reductase Genes on Methotrexate-Induced Cytotoxicity

윤성애,최정란,김정오,신정영,XiangHua Zhang,강진형 대한암학회 2010 Cancer Research and Treatment Vol.42 No.3

Purpose The aim of this study is to investigate the effect of genetic variations and the expression of the reduced folate carrier (RFC) and dihydrofolate reductase (DHFR) on the drug sensitivity to methotrexate (MTX) in different cancer cell lines. Materials and Methods We examined the six human cancer cell lines (MCF-7, AGS, A549, NCI-H23, HCT-116 and Saos- 2). The cytotoxicity of MTX was measured by sulforhodamine B (SRB) assay. The expressions of the DHFR and RFC were evaluated by real-time PCR and western blotting. Four single nucleotide polymorphisms (SNPs) of the DHFR and two SNPs of the RFC were genotyped. Results The IC50s of MTX was in an extensively broad range from 6.05±0.81 nM to>1,000 nM in the cell lines. The Saos-2 (>1,000 nM) and MCF-7 (114.31±5.34 nM) cells were most resistant to MTX; in contrast, the AGS and HCT-116 cells were highly sensitive to MTX with an IC50 of 6.05±0.81 nM and 13.56±3.76 nM, respectively. A reciprocal change of the RFC and DHFR mRNA expression was found between the MTX-sensitive AGS and MTX-resistant Saos-2 cells. There was no significant difference in the expression levels of RFC protein in both the AGS and Saos-2 cells, whereas DHFR protein was more increased in the MTX-resistant Saos-2 cells treated with MTX. The genotype of the MTX-sensitive AGS cells were mutant variants of the DHFR; in contrast, the Saos-2 cells had the wild-type of the DHFR. Conclusion In conclusion, this study showed that inverse change of the RFC and DHFR mRNA and protein expression was associated with RFC and DHFR polymorphisms and it is postulated that this phenomenon might play an important role in sensitivity of certain cancers to MTX.

급성 Cyclosporin A 신독성 쥐의 신장에서 칼슘 유도성 사립체 종창의 억제

윤성애,방병기 가톨릭대학 (가톨릭신학대학) 대학원 1996 가톨릭大學醫學部 論文集 Vol.49 No.2

보문 : 섬강수계 부착규조의 유입능 특성: 자연기질과 인공기질의 비교

윤성애 ( Sung Ae Yoon ),김난영 ( Nan Young Kim ),변명섭 ( Myeong Seop Byoen ),김백호 ( Baik Ho Kim ),황순진 ( Soon Jin Hwang ) 한국하천호수학회 2009 생태와 환경 Vol.42 No.1

저온기 부영양 수계 규조군집의 유입능

윤성애 ( Sung Ae Yoon ),긴난영 ( Nan Young Kim ),김백호 ( Baik Ho Kim ),황순진 ( Soon Jin Hwang ) 한국하천호수학회 2008 생태와 환경 Vol.41 No.3

보문 : 도시 하천(원주천) 유입이 섬강 하류 부착규조 군집에 미치는 영향

윤성애 ( Sung Ae Yoon ),김난영 ( Nan Young Kim ),김백호 ( Baik Ho Kim ),황순진 ( Soon Jin Hwang ) 한국하천호수학회(구 한국육수학회) 2010 생태와 환경 Vol.43 No.2

보문 : 인공수로에서 고농도 탁수가 수질 및 부착 규조류 군집에 미치는 영향

윤성애 ( Sung Ae Yoon ),유경아 ( Kyung A You ),박지형 ( Ji Hyoung Park ),김백호 ( Baik Ho Kim ),황순진 ( Soon Jin Hwang ) 한국하천호수학회(구 한국육수학회) 2011 생태와 환경 Vol.44 No.1

We examined the effect of the turbid water on the periphytic diatom community in an artificial stream system. The artificial stream was constructed with transparent acryl and composed of four channels. Each channel (20cm×200cm×40 cm) was supplied continuously with eutrophic lake water. In order to the freely colonize and grow diatoms, artificial substrate was installed with commercial slide glass soaked in 1% agar. Prior to introducing turbid water, the artificial stream was operated with lake water for 6 days to permit the propagation of diatom community on the substrates. The turbid water prepared with sediment sieved with Φ 64μm at 2g L-1 (final concentration, 300 NTU) was provided daily for 50 minute duration. The experiment was conducted for 7 days with manipulated experimental condition of light (50~80μmol m-2 s-1, light : dark=24: 0), temperature (10±1℃), and flow rate (0.31cm s-1). Sampling and analysis were conducted daily for water quality and diatom. Turbidity of the water varied 162.2~173.2 NTU during the experiment. After introduction of turbid water, DO, pH and TN were decreased, while SS and TP increased significantly. A total of 14 genera and 47 species of diatoms was observed on the artificial substrates during the experimental period. Of these, Navicula appeared to be a most dominant genus with 10 species, followed by Cymbella (6 species), Fragilaria (6 species) and Gomphonema (5 species). Achnanthes minutissima was the most dominant species (>70% of total frequency) in both control and treatment experiments. Increase in diatom abundance lasted for three days since turbid water introduction, after that they gradually decreased by the termination of the experiment. These results suggest that frequent supply of highly-concentrated turbid water significantly decreases the periphytic diatom community, and retard the recovery of the stable food-web within the stream.

Association of Genetic Variations with Pemetrexed-Induced Cytotoxicity in Non-Small Cell Lung Cancer Cells

Seong-Ae Yoon(윤성애),Jung Ran Choi(최정란),Jeong-Oh Kim(김정오),Jung-Young Shin(신정영),XiangHua Zhang(장향화),Jin-Hyoung Kang(강진형) 한국생명과학회 2010 생명과학회지 Vol.20 No.1

페메트렉시드(pemetrexed, alimta<SUP>®</SUP>)는 중피종(mesothelioma)과 비소세포폐암 (non-small cell lung cancer)을 비롯한 다양한 암종에서 엽산(folate) 대사과정에 관여하는 대사물질의 활성을 억제하여 항암효능을 나타낸다. 다중표적 항암제 (multitargeted antifolate)인 pemetrexed는 엽산의 세포내 주요 이동통로인 reduced folate carrier(RFC)를 통해 세포 내로 유입된 후 folylpolyglutamate synthetase (FPGS)에 의해 폴리글루타민산염(polyglutamate) 유도체로 활성되고 thymidylate synthase (TS)와 dihydrofolate reductase (DHFR)를 표적하는 것으로 알려져 있다. 조직형이 서로 다른 비소세포폐암 세포주를 선정하여 pemetrexed의 대사과정에 관여하는 유전자들의 단일염기서열 다형성을 조사하고, mRNA와 단백질의 발현 정도를 비교하여 pemetrexed의 세포독성 효과와의 상관성을 분석하였다. 4개의 비소세포폐암 세포주인 A549, PC14, HCC-1588과 H226에서 RFC, FPGS, TS와 DHFR의 유전형을 조사하였다. Pemetrexed의 약물의 감수성을 알아보기 위해 real-time PCR과 Western blot 방법으로 mRNA 발현과 단백질 발현 정도를 비교하였고, SRB 법으로 약물에 대한 세포독성 효과를 측정했다. PC14 세포주와 H226 세포주에서는 약물처리 전 RFC와 FPGS의 mRNA 발현이 높은 것으로 나타났고, IC50값이 각각 0.08±0.01 uM과 0.07±0.01 uM로 pemetrexed에 대한 감수성이 높은 것을 알 수 있었다. A549 세포주에서 TS의 유전형이 2R/2R일 때 mRNA발현이 증가하고 pemetrexed의 약물 저항성과 관련이 있었다. 반면, TS의 유전형이 3R/3R로 나타난 H226에서는 mRNA 발현이 낮은 것을 알 수 있었지만 pemetrexed의 높은 감수성과 관련이 있었다. 세포주 모두에서 pemetrexed 약물처리 후 DHFR의 mRNA 발현은 약물처리 전보다 낮아지는 경향을 보였지만 단백질 발현은 오히려 증가하는 상반된 결과를 보였다. 또한 DHFR 프로모터에 위치한 -1726C>T, -1188A>C SNP는 서로 연쇄 불평형 상태(linkage disequilibrium, LD)에 있었다. 연구결과에서 pemetrexed의 세포독성 효과는 약물 대사과정에 관여하는 여러 분자들의 유전형과 발현 정도에 의해 결정되는 것을 알 수 있었고, 다양한 분석결과를 토대로 항암효능을 평가하는 것이 필요하다고 생각된다. Pemetrexed has demonstrated clinical activity in non-small cell lung cancer (NSCLC) as well as other solid tumors. It transports into the cells via reduced folate carrier (RFC) and is polyglutamated by folypolyglutamate synthetase (FPGS). Pemetrexed directly inhibits several folate-dependent enzymes such as thymidylate synthase (TS), dihydrofolate reductase (DHFR), and glycinamide ribonucleotide formyltransferase (GARFT). We investigated the effects of genetic variations and the expression of RFC, FPGS, TS and DHFR enzymes on drug sensitivity to pemetrexed in NSCLC cells. Polymorphisms in RFC, FPGS, and DHFR were genotyped in four NSCLC cells - A549, PC14, HCC-1588, and H226. Real-time RT-PCR and Western blot was performed to evaluate mRNA transcripts and protein of these genes. The cytotoxicity of pemetrexed was measured by SRB assay. In PC14 and H226 cells, increased mRNA expressions of RFC and FPGS were associated with higher cytotoxicity to pemetrexed. 2R/2R genotype of TS and its increased mRNA expression were associated with drug resistance to pemetrexed in A549 cells, whereas 3R/3R genotype in TS with decreased mRNA expression was associated with higher sensitivity in H226 cells. After pemetrexed treatment, an inverse change of DHFR mRNA and protein expression was found. The strongest linkage disequilibrium (LD) was discovered between-1726C>T and -1188A>C SNP of DHFR gene. Our findings suggest the cytotoxic effect of pemetrexed may be associated with genetic polymorphisms and the expression level of genes involved in pemetrexed metabolisms in NSCLC cells.

대학 AV Center의 문제점 분석 및 그 해결방안에 관한 연구

오성숙,윤성애 이화여자대학교 사범대학 시청각교육과 1975 시청각 교육 Vol.- No.9

李朝女人特殊外出服薦衣에 對한 調査 및 擬製

徐淑子,朴今順,尹聖愛,金外泰,金槿,許鎬京,宋明淑 曉星女子大學家政學會 1976 효대가정학회지 : 효성여대 Vol.6 No.1

When women of the Lee Dynasty went out doors, regardless of high or low class, they used to wear various kinds of long hoods to hide their faces. This long hood was designed differently by their social positions or the times. The Confucianism thought that "a boy and a girl should not sit together after they have reached the age on seven," and the historical background of the times that the social position of women was neglected forced them to be confined in their houses. This custom continued for a long time. Of course, considering that the custom was not only in Korea but also in such Middle East countries as Iraq, Afghanistan, Saracen, we don't know there are any traces of outside influence on each other, indeed it is a very interesting manner. In Seoul and its neighboring districts "jang-ot' was used to hide the face of a woman, while in Gwanbuk and Yeongnam district "cheonai" was used. The long hood was different according to it's district. Though in 1975 a special class activity studying clothing in the Home Economomic department planned to make an investigation into the special street dress of Korea. They couldn't do it because of problem of expence. After we investigated "cheonai" which was worn habitually by women of Yeongnam district, we decided to make an imitation of it. We made "cheonai." which Andong-born Mrs. You, aged around 80 wore when young, an object of our study and created an imitation of it. The results of our research appear in the measurements of the clothes and the art of stitchery used on the clothing.

대형 배양장치에서 기능그룹에 기초한 식물플랑크톤 천이 특성

이경락,노성유,이재윤,윤성애,이재학,신유나,이수웅,류덕희,이재관 한국하천호수학회 2017 생태와 환경 Vol.50 No.4

The present study was conducted from August to December 2016 in a cylindrical water tank with a diameter of 1 m, a height of 4 m and a capacity of 3,000 L. The field water and sediment from the Nakdong River were also sampled for the experimental culture (field water+sediment) and control culture (field water), respectively. In this study, we aimed to investigate phytoplankton succession pattern using the phytoplankton functional group in the enclosed culture system. A total of 50 species in 27 genera including Chlorophyceae (30 species), Bacillariophyceae (11 species), Cyanophyceae (7 species), and Cryptophyceae (2 species) were identified in the experimental and control culture systems. A total of 19 phytoplankton functional groups (PFGs) were identified, and these groups include B, C, D, F, G, H1, J, K, Lo, M, MP, N, P, S1, TB, W0, X1, X2 and Y. In particular, W0, J and M groups exhibited the marked succession in the experimental culture system with higher biovolumes compared to those of the control culture system, which may be related to the internal cycling of nutrients by sediment in the experimental culture system. The principal component analyses demonstrated that succession patterns in PFG were associated with the main environmental factors such as nutrients (N, P), water temperature and light intensity in two culture systems. In conclusion, the present study showed the potential applicability of the functional group for understanding the adaptation strategies and ecological traits of the phytoplankton succession in the water bodies of Korea.