HUMAN PROSTATIC 3A HYDROXYSTEROID DEHYDROGENASE
HUMAN PROSTATIC 3A HYDROXYSTEROID DEHYDROGENASE
批准号:
2614194
负责人:
HSUEH-KUNG LIN
金额:
$2.24万
依托单位国家:
美国
项目类别:
财政年份:
1998
资助国家:
美国
项目状态:
已结题
起止时间:
1998-09-28 至 1998-11-30
关键词:
RNase protection assay androgens cell line cell proliferation enzyme activity epidermal growth factor genetic regulation genetic regulatory element genetic transcription growth factor receptors hormone regulation /control mechanism hydroxysteroid dehydrogenases isozymes messenger RNA nucleic acid sequence prostate protein structure function steroid hormone biosynthesis
中文摘要
描述(改编自申请人的摘要):雄激素目标
组织中的 3a-羟基类固醇脱氢酶 (3a-HSD) 可能调节
通过相互转化占据雄激素受体(AR)
5a-二氢睾酮(5a-DHT,一种强效雄激素)
5a-androstane-3a,17b-diol(3a-二醇,一种弱雄激素)。 我们已经获得了类型
2 型和 3 型 3a-HSD cDNA 在人前列腺中表达并过表达
大肠杆菌中的酶。 这些重组酶的动力学研究表明
3 型 3a-HSD 同时充当 3a-和 17b-HSD 来灭活活性
雄激素,而 3 型 3a-HSD 可将 5a-DHT 与 3a- 二醇相互转化。
前列腺原代培养物中 3a-HSD mRNA 水平较高
上皮细胞多于基质细胞; 3a-HSD mRNA 水平升高
在源自良性的上皮细胞的原代培养物中观察到
前列腺增生和前列腺癌组织。 稳定的表达
3a-HSD mRNA 的状态水平通过表皮生长 (EGF) 上调
人前列腺细胞系、LNCaP(雄激素敏感)和 PC3(雄激素
不敏感)。
该提案的重点是检查类型的生理功能
2型和3型3a-HSD调节雄激素代谢及其活性
将研究调节前列腺细胞增殖的作用。 这将
通过将 2 型和 3 型 3a-HSD cDNA 稳定转染至
这些细胞。 二、内源性2型和3型3a-HSD水平
将在从细胞系和正常细胞中提取的 RNA 中检查转录本
使用核糖核酸酶保护测定(RPA)检测前列腺。 三、EGF调节
将使用 RPA 检查 2 型和 3 型 3a-HSD mRNA 水平。 变化
通过 EGF 介导的 3a-HSD 表达将在细胞裂解物中进行检查
酶活性的免疫滴定。 四、要了解
组成型和 EGF 调节的 2 型和 3 型 3a-HSD 表达
前列腺细胞中,3a-HSD 基因的 5' 侧翼区域将是
负责转录调控的测序和顺式作用元件
将鉴定出两种亚型。
英文摘要
DESCRIPTION (Adapted from the Applicant's Abstract): In androgen target
tissues 3a-hydroxysteroid dehydrogenases (3a-HSDs) may regulate the
occupancy of androgen receptor (AR) by interconverting
5a-dihydrotestosterone (5a-DHT, a potent androgen) with
5a-androstane-3a,17b-diol (3a-diol, a weak androgen). We have obtained type
2 and type 3 3a-HSD cDNAs expressed in human prostate and overexpressed the
enzymes in E. coli. Kinetic studies of these recombinant enzymes show that
type 3 3a-HSD functions as both a 3a-and 17b-HSD to inactivate active
androgens, whereas, type 3 3a-HSD interconverts 5a-DHT with 3a- diol.
Levels of 3a-HSD mRNA were higher in primary cultures of prostatic
epithelial cells than stromal cells; and elevated levels of 3a-HSD mRNA were
observed in primary cultures of epithelial cells derived from benign
prostatic hyperplasia and prostatic carcinoma tissues. Expression of steady
state levels of 3a-HSD mRNA is up-regulated by epidermal growth (EGF) in
human prostatic cell lines, LNCaP (androgen sensitive) and PC3 (androgen
insensitive).
The focus of this proposal is to examine the physiological functions of type
2 and type 3 3a-HSD in regulating androgen metabolism and their activities
in modulating prostatic cell proliferation will be investigated. This will
be accomplished by stably transfecting type 2 and type 3 3a-HSD cDNAs into
these cells. Second, levels of endogenous type 2 and type 3 3a-HSD
transcripts will be examined in RNA extracted from the cell lines and normal
prostate using ribonuclease protection assay (RPA). Third, EGF-regulated
type 2 and type 3 3a-HSD mRNA levels will be examined using RPA. Changes in
3a-HSD expression mediated by EGF will be examined in cell lysates by
immunotitration of the enzyme activity. Fourth, to understand the
constitutive and EGF- regulated type 2 and type 3 3a-HSD expression in
prostatic cells, the 5'-flanking regions of the 3a-HSD genes will be
sequenced and cis-acting elements responsible for transcription regulation
of two isoforms will be identified.
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HUMAN PROSTATIC 3A HYDROXYSTEROID DEHYDROGENASE
-
批准号:6523761
-
项目类别:
-
资助金额:$14.55万
-
财政年份:1998
-
负责人:HSUEH-KUNG LIN
-
依托单位:
HUMAN PROSTATIC 3A HYDROXYSTEROID DEHYDROGENASE
-
批准号:6177945
-
项目类别:
-
资助金额:$6.6万
-
财政年份:1998
-
负责人:HSUEH-KUNG LIN
-
依托单位:
HUMAN PROSTATIC 3A HYDROXYSTEROID DEHYDROGENASE
-
批准号:2906335
-
项目类别:
-
资助金额:$15.6万
-
财政年份:1998
-
负责人:HSUEH-KUNG LIN
-
依托单位:
HUMAN PROSTATIC 3A HYDROXYSTEROID DEHYDROGENASE
-
批准号:6381385
-
项目类别:
-
资助金额:$14.55万
-
财政年份:1998
-
负责人:HSUEH-KUNG LIN
-
依托单位:
HUMAN PROSTATIC 3A HYDROXYSTEROID DEHYDROGENASE
-
批准号:6405358
-
项目类别:
-
资助金额:$9.0万
-
财政年份:1998
-
负责人:HSUEH-KUNG LIN
-
依托单位:
HUMAN PROSTATIC 3A HYDROXYSTEROID DEHYDROGENASE
-
批准号:6071811
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项目类别:
-
资助金额:$13.71万
-
财政年份:1998
-
负责人:HSUEH-KUNG LIN
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依托单位:
海外基金