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Transcriptional Activation and Repression of Cell Cycle Regulatory Molecules by Trichostatin A
백종수,이희경,조영수,김성영,박관규,장영채,Baek Jong-Soo,Lee Hee-Kyung,Cho Young-Su,Kim Sung-Young,Park Kwan-Kyu,Chang Young-Chae Korean Society of Life Science 2005 생명과학회지 Vol.15 No.6
Dihydrofolate reductase (dhfr) promoter에는 전사 인자 Spl과 E2F가 결합하는 cis-acting 배열을 가지고 있다. dhfr 유전자의 전사는 세포 주기 Gl/S기 동안 최대의 발현을 나타낸다. 또한 Spl 전사 인자는 dhfr 유전자 발현의 활성화 및 불활성화를 조절하는 다양한 역할에 대한 연구가보고 되고 있으며, 최근 Spl-Rb과 E2F4-pl30 복합체가 CHOC400 세포에서 dhfr 유전자 발현에 안정한 형태를 형성하여 dhfr 발현을 억제한다는 연구 결과가 보고되었다. 본 연구에서는 Rb-양성 골육종 세포인 U2OS 및 Rb음성인 자궁경부암 C33A 세포에서 histon deacetylase (HDAC)에 대한 특이적인 저해제인 trichostatn A (TSA)를 처리한 후 세포주기 조절에 중심적 인자들인 dhfr cyclin E 및 cyclin A의 전사활성에 대한 HDACl의 기능을 조사하였다. U2OS 및 C33A 세포에서 TSA를 처리한 후, dhfr, cyclin E, cyclin A에 대한 mRNA 및 단백질 발현을 조사한 결과 U2OS 세포 특이적으로 dhfr cyclin E의 mRNA 발현과 단백질 발현이 크게 증가하였지만, cyclin A의 발현은 감소하였다. U2OS 세포에서 dhfr promoter construct에 대한 전사활성을 검사한 결과, TSA 처리는 dhfr promoter 영역으로부터 E2F 결합부위를 제거시킨 DHFR-Spl-luc를 통하여 dhfr promoter활성이 약 14배 증가되었다, 그러나 dhfr promoter 영역으로부터 Spl 결합부위를 제거시킨 DHFR-E2F-luc 영역을 포함하고 있는 promoter 활성은 TSA 처리에 의해 크게 증가되지 않았다. 본 연구에서 이러한 결과는 HDACI이 Spl을 통하여 dhfr promoter활성을 제어한다는 사실을 입증하였다. 한편 TSA는 U2OS 세포에서 HDAC의 활성을 통해서 세포주기 관련 인자들 가운데서 Gl 후기부터 활성화되는 대표적인 인자들인 dhfr과 cyclin E의 발현을 증가시키지만 G2 기에서 활성화되는 대표적인 인자인 cyclin A의 발현을 억제하는 상반된 기능을 가지고 있다는 사실을 확인하였다. The dihydrofolate reductase (dhfr) promoter contains cis-acting element for the transcription factors Spl and E2F. Transcription of dhfr gene shows maximal activity during the Gl/S phase of cell cycle. The member of the Spl transcriptional factor family can act as both negative and positive regulators of gene expression. There was a report that Spl-Rb and E2F4-pl30 complexes cooperate to establish stable repression of dhfr gene expression in CHOC400 cells. Here, we examined the role of HDAC in dhfr, cyclin E, and cyclin A gene regulation using the histone deacetylation inhibitor, trichostatin A (TSA) in U2OS and C33A cells, a Rb-positive human osteosarcoma cell line, and a Rb-negative cervical carcinoma cell line, respectively. When the dhfr promoter constructs were applied in U2OS cells, TSA markedly stimulated over 14-fold of dhfr promoter activity through dhfr-Spl sites by the deletion of an E2F element. In contrast, the deletion of dhfr-Spl binding sites completely abolished promoter stimulation by TSA. The dhfr promoter activity including dhfr-Spl sites increased only 2-fold in C33A cells. Promoter activity containing only dhfr-E2F site did not have much effect by the treatment of TSA in both U2OS and C33A cells. On the other hand, treatment with TSA induced significantly mRNA expression of dhfr and cyclin E, whereas levels of cyclin A decreased in U2OS cells, but had no effect in C33A cells. These results indicate that TSA have contradictory effect, activation of dhfr and cyclin E genes on Gl phase, and down-regulation of cyclin A on G2 phase through transcriptional regulation in U2OS cells.
복합운동이 복부비만 여성의 HOMA index와 Ghrelin에 미치는 영향
김동희(Dong Hee Kim),백종수(Jong Soo Baek),조성일(Sung Il Cho),이하얀(Ha Yan Lee) 한국발육발달학회 2006 한국발육발달학회지 Vol.14 No.3
The purpose of this study was to investigate the effects of combined exercise on the fasting blood glucose, insulin, HOMA index, and ghrelin. The subjects for this study were sixteen abdominal obesity women(waist circumference = over 80㎝) and divided into 2 groups: combined exercise(aerobic+resistance exercise) group(n=8), and control group(n=8). The aerobic exercise(running) performed four times a week at the intensity of 50~75%HRmax(1~5 weeks; 50~60%HRmax, 6~10 weeks; 61~75% HRmax). The resistance exercise(behind squat, sit-up, bench press, full-up, arm curl) performed four times a week at the intensity of 50~75%HRmax(1~5 weeks; 1RM, 60%HRmax, 6~10 weeks; 1RM, 75%HRmax). And the change of fasting blood glucose, insulin, HOMA index, and ghrelin have been measured before and after 10 weeks of exercise respectively. Statistical techniques for data analysis were two-way ANOVA to determine the difference between before and after 10 weeks of exercise and the difference between the combined exercise group and the control exercise group. The 5% level of significance was utilized as the critical level for acceptance of hypotheses for the study. The following results were obtained from this study. 1. The change of fasting blood glucose, insulin, HOMA index in the combined exercise group was decreased after 10 weeks of exercise. And the change of fasting blood glucose was statistical significant difference between the groups(p<.05). 2. The change of ghrelin in the combined exercise group was statistical significant increased after 10 weeks of exercise. And there was no significant difference between the groups, but there was significant difference between the weeks and group×week interaction(p<.05).
중년여성의 16주간 요가운동이 항산화효소와 과산화지질에 미치는 영향
김동희(Dong Hee Kim),박해선(Hae Sun Park),백종수(Jong Soo Baek),이하얀(Ha Yan Lee),김회원(Hoe Won Kim),신세훈(Se Hoon Shin),장선웅(Seon Woong Jang) 한국사회체육학회 2007 한국사회체육학회지 Vol.0 No.31
The purpose of this study was to analyze the effects of yoga exercise on the activation of antioxidation enzyme and lipid peroxidation. The subjects for this study were eight women. Yoga exercise program consisted of 28 item, performed for 60 minutes in a bout, 5 times a week for 16 weeks. And the change of the activation of antioxidation enzyme(SOD, CAT, GPx), lipid peroxidation(MDA) have been measured before and post 10 weeks respectively. The results obtained from this study were as follows; 1. CAT(p<.01) and GPx(p<.05) were increased statistic significantly after 16 weeks of yoga exercise. But SOD was increased after 16 weeks of yoga exercise. 2. MDA was decrease after 16 weeks of yoga exercise.