Analysis of estrogen-induced genes in breast cancer

乳腺癌雌激素诱导基因分析

基本信息

批准号：
06454359
负责人：
IINO Yuichi
金额：
$ 4.1万
依托单位：
Gunma University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1996
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06454359/
关键词：
Breast cancer Estrogen Choline kinase PCR Cloning

项目摘要

We found that estrogen increased the activity and protein levels of choline kinase in DMBA-induced rat mammary carcinoma. Choline kinase gene was expected to be regulated by estrogen or by factor induced by estrogen. To investigate the activation mechanism of the enzyme, we carried out two experiments. First, we isolated and characterized rat choline kinase gene. The 5'region of choline kinase gene showed features not only of a typical housekeeping gene but also of a gene regulated by a various kind of regulatory factors. In fact, the carcinogen 3-methylcholanthrene, and hepatotoxic carbon tetrachloride induced this enzyme in rat liver. However, neither estrogen responsive element norits similar sequence of the estrogen binds was found between 5'region and the beginning of the first intron of this gene. Second, we obtained cDNAs from mRNA induced by estrogen in DMBA-induced rat mammary carcinoma and in human breast cancer cell line MCF-7 implanted in nude mice. Total RNA was extracted from these tumors in the presence or absence of estrogen. cDNA was synthesized by reverse transcription and used to polymerase chain reaction with a various pair of primers. Reaction products were separated by electrophoresis and differentially displayd in parallel. We obtained 13 cDNAs of estrogen-induced genes from rat tumors and 39 cDNAs of these from MCF-7 tumors. We are analyzing these cDNAs. We will elucidate the mechanism of choline kinase activation and growth regulation by estrogen.

我们发现雌激素增加了DMBA诱导的大鼠乳腺癌中胆碱激酶的活性和蛋白质水平。预计胆碱激酶基因将由雌激素或雌激素诱导的因子调节。为了研究酶的激活机制，我们进行了两个实验。首先，我们分离了大鼠胆碱激酶基因。胆碱激酶基因的5'区域不仅显示出典型的管家基因的特征，而且表现出受各种调节因子调节的基因。实际上，致癌3-甲基胆碱和肝毒性四氯化碳诱导大鼠肝脏中的这种酶。然而，在5'Rigion和该基因的第一个内含子的开始之间，雌激素反应元件均未发现雌激素结合的相似序列。其次，我们从DMBA诱导的大鼠乳腺癌和植入裸鼠的人类乳腺癌细胞系MCF-7中从雌激素诱导的mRNA中获得了cDNA。在存在或不存在雌激素的情况下，从这些肿瘤中提取总RNA。通过逆转录合成cDNA，并用于与各对引物的聚合酶链反应。通过电泳分离反应产物，并平行显示差异显示。我们从MCF-7肿瘤中获得了来自大鼠肿瘤的雌激素诱导基因的13个cDNA，其中39个cDNA。我们正在分析这些CDNA。我们将阐明雌激素的胆碱激酶激活和生长调节的机理。