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完整後設資料紀錄
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dc.contributor.advisor郭應誠
dc.contributor.authorCheng-Fen laien
dc.contributor.author賴政分zh_TW
dc.date.accessioned2021-06-13T06:57:40Z-
dc.date.available2005-08-01
dc.date.copyright2005-08-01
dc.date.issued2005
dc.date.submitted2005-07-28
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dc.identifier.urihttp://tdr.lib.ntu.edu.tw/jspui/handle/123456789/35543-
dc.description.abstract類固醇轉錄因子(Steroidogenic Factor1, SF-1)是孤核受體(orphan nuclear receptor)的一種,能調控許多類固醇生成酵素基因的表現,對腦下垂體、下視丘、腎上腺及性線的發育及分化非常重要。在小鼠與人的腎上腺皮質腫瘤細胞內(Y1 and H295 cells),使用組蛋白去乙醯基轉移酵素抑制劑 ,如Trichostatin A和Sodium butyrate,能增加Histone 4的乙醯化 (Acetylation),卻減少SF-1與其下游膽固醇側鏈分切酵素(P450scc)的表現,這個抑制現象與劑量和作用時間成正比。報導基因表現分析的實驗中,TSA抑制了SF-1啟動子的活性;TSA能增加外源性SF-1蛋白質的表現,但減少內源性SF-1蛋白質的表現量。表示HDAC 抑制劑是透過SF-1啟動子來控制基因轉錄的表現。許多文獻指出,SF-1啟動子上面,具有一個重要的調控序列,E-box,與其結合的兩個主要蛋白質為上游刺激因子一及二 (Upstream Stimulatory Factor 1 and USF2)。膠體電泳位移分析 (EMSA) 結果表示,HDAC 抑制劑同時減少SF-1啟動子上調控單位(E box and CCAAT box)與其轉錄因子間的結合,也減少了USF1和USF2蛋白質的表現量。TSA抑制了P450scc及USF2的mRNA,但卻增加了USF1的mRNA;此結果與減少的USF1蛋白質相矛盾,故使用HDAC 抑制劑可能會影響特定蛋白質的穩定性並加速其降解。zh_TW
dc.description.abstractThe orphan nuclear receptor steroidogenic factor 1 (SF-1, A4BP, or NR5A1) is a key transcription factor that regulates the expression of many steroidogenic enzymes. It has also been demonstrated to play an important role in the development and differentiation of hypothalamus, pituitary, adrenal glands, and gonads. Being such an important factor, the regulation of SF-1 expression is highly investigated. We used inhibitors of histone deacetylases which generally activate gene expression and used as anticancer drugs such as trichostatin A and sodium butyrate. Histone deacetylase inhibitors although increased the acetylation of Histone 4, reduced the protein content of SF1 and cytochrome P450 side-chain cleavage enzyme, a SF-1 controlled gene, in a dose and time-dependent manner in adrenocorticol cells, Y1 and H295 cells. In a reporter assay, TSA reduced the SF-1 promoter activity. In Y1 stable clone, which expresses SF-1-HA driven by CMV promoter, TSA increased exogenous SF-1-HA protein, however, reduced endogenous SF-1 protein. Both results indicate that SF-1 is transcriptionally down-regulated by these inhibitors. TSA reduced binding of E-box and CCAAT box binding factor to SF-1 promoter in Electorphoretic Mobility Shift Assay. The E-box binding proteins, USF1 (upstream stimulatory factor 1) and USF2 were down-regulated by TSA as well as SF-1. These results suggest that TSA-inhibited expression of SF-1 may provide insight to uncover the regulatory mechanism of SF-1 gene. Interesting, we found that TSA treatment increases mRNA of SCC and USF2 but reduces USF1 mRNA in Quantitative-Real-Time PCR. Increased USF1 mRNA and reduced USF1 protein content after TSA treatment provides a new association of protein stability and acetylation.en
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Previous issue date: 2005
en
dc.description.tableofcontentsTable of Contents
Abbreviation 1
Abstract in Chinese 2
Abstract 3
Introduction 4
I、Steroid hormones 4
Classes & Functions 4
CYP11A1 (P450scc) 4
II、Steroidogenic Factor 1 5
Characterization 5
Structure 5
Regulation 6

III、Transcriptional regulation 7
Chromatin structure & Histone modification 7
Acetylation & Histone acetyltransferase 8
Dacetylation & Histone deacetylase 9

IV、HDAC inhibitors 9
Trichostatin A 10
Sodium Butyrate 10
V、HDAC inhibitors down regulate gene expression 10
VI、HDAC inhibitors inhibit SF-1 expression 10
In a dose- and time-dependent manner 11
Reduces SF-1-BS, E-box and CCAAT box binding 11
Reduces binding of USF1 and USF2 to E-box 12

Aim 13
Material and Methods 14
Cell culture and transfection 14
Plasmids construction 14
Western Blot analysis 15
Electrophoretic Mobility Shift Assay (EMSA) 15
Real-Time PCR 16
Administration of Trichostatin A and Sodium Butyrate 16
Results 18
I. HDAC inhibitors reduce SF-1 expression through SF-1 promoter region 18
II. TSA reduces SCC and USF2 mRNA but increases USF1 mRNA level 18
III. HDAC inhibitor decreases USF1 protein stability 19
IV. HDAC inhibitor effects in steroidogenesis in mice 19
Discussions 20
I. USF1 and USF2 have different response to TSA 20
The regulation of mRNA expression 20
The regulation of protein content 21
II. HDAC inhibitor effects in vivo 23
References 25
Table and Figures 33
dc.language.isoen
dc.subject因子zh_TW
dc.subject上游刺激因子一及二zh_TW
dc.subject膽固醇側鏈分切酵素zh_TW
dc.subject乙醯化zh_TW
dc.subject組蛋白去乙醯基轉移酵素抑制劑zh_TW
dc.subject腎上腺皮質腫瘤細胞zh_TW
dc.subject孤核受體zh_TW
dc.subject固醇轉錄zh_TW
dc.subjectthe cholesterol side-chain cleavage enzymeen
dc.subjecttrichostatin Aen
dc.subjecthistone acetylatransferaseen
dc.subjectP450 side chain cleavage enzymeen
dc.subjectsteroidogenic factor 1en
dc.title組蛋白去乙醯基轉移酵素阻礙劑抑制腎上腺皮質細胞內類固醇轉錄因子的表現zh_TW
dc.titleInhibitors of Histone Deacetylase Down-regulate the Expression of Steroidogenic Factor 1 in Adrenocorticol Cellsen
dc.typeThesis
dc.date.schoolyear93-2
dc.description.degree碩士
dc.contributor.oralexamcommittee鍾邦柱,詹東榮,阮麗蓉,楊文明
dc.subject.keyword類,固醇轉錄,因子,孤核受體,腎上腺皮質腫瘤細胞,組蛋白去乙醯基轉移酵素抑制劑,乙醯化,膽固醇側鏈分切酵素,上游刺激因子一及二,zh_TW
dc.subject.keywordhistone acetylatransferase,P450 side chain cleavage enzyme,the cholesterol side-chain cleavage enzyme,steroidogenic factor 1,trichostatin A,en
dc.relation.page45
dc.rights.note有償授權
dc.date.accepted2005-07-28
dc.contributor.author-college生物資源暨農學院zh_TW
dc.contributor.author-dept獸醫學研究所zh_TW
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