人HDAC1表达载体的构建及其对ERa和ERβ蛋白水平的调控

发布时间：2018-01-09 07:09

本文关键词：人HDAC1表达载体的构建及其对ERa和ERβ蛋白水平的调控　出处：《郑州大学》2006年硕士论文　论文类型：学位论文

更多相关文章： HDAC1 ERα ERβ 克隆与表达 蛋白水平

【摘要】：乳腺癌是妇女最常见的恶性肿瘤之一。雌激素(estrogen)对于维持正常乳腺组织的发育和功能至关重要，雌激素作用的发挥需要通过与雌激素受体α和β(ERα和ERβ)的结合，而ERα和ERβ二者又都可以以很高的活性与各种组蛋白结合，而组蛋白的乙酰化是影响细胞各种生理功能的最典型的一种方式，它依赖于组蛋白乙酰化酶(HATs)和组蛋白去乙酰化酶(HDACs)两类酶。在乳腺细胞中，ER正是通过募集如HATs和HDACs等转录共调节因子，利用它们呈现的各种酶活性来调控基因的表达。目的本文意在通过构建可在真核细胞中表达的重组质粒pcDNA3-HA-hdacl，，研究组蛋白去乙酰化酶1(HDAC1)和雌激素受体ERα和ERβ在体外的相互结合作用，以及HDAC1对ERα和ERβ蛋白水平的影响规律，从而在一定角度揭示乳腺细胞的增殖、分化和癌变的过程，为更好地寻找治疗癌症的药物靶标打下基础。方法 ①从癌细胞中用TRIzol一步法提取总RNA产物； ②用Promega反转录试剂盒进行反转录合成cDNA； ③利用PCR技术扩增出hdacl基因完整编码区序列； ④通过DNA重组技术构建含hdacl片段的真核表达载体pcDNA3-HA-
[Abstract]:Breast cancer is one of the most common malignant tumors in women. Estrogen estrogen is essential to maintain the development and function of normal breast tissues. Estrogen plays a role in binding to estrogen receptor 伪 and 尾 -ER 伪 and ER 尾, and both ER 伪 and ER 尾 can bind to various histones with high activity. Acetylation of histone is the most typical way to affect various physiological functions of cells. It depends on histone acetylase (HATs) and histone deacetylase (HDACs) in breast cells. ER regulates gene expression by recruiting transcriptional co-regulatory factors such as HATs and HDACs. Purpose The purpose of this study is to construct a recombinant plasmid pcDNA3-HA-hdacl that can be expressed in eukaryotic cells. Study group protein deacetylase 1 (HDAC1) and estrogen receptor ER 伪 and ER 尾 binding in vitro, and the effect of HDAC1 on ER 伪 and ER 尾 protein levels. Thus, the process of proliferation, differentiation and canceration of breast cells was revealed in a certain angle, which laid a foundation for better searching for drug targets for cancer treatment. Method 1Total RNA products were extracted from cancer cells by TRIzol one-step method. (2) reverse transcription synthesis of cDNAusing Promega reverse transcription kit; (3) the complete coding region of hdacl gene was amplified by PCR. Construction of eukaryotic expression vector pcDNA3-HA- containing hdacl fragment by DNA recombination technique
【学位授予单位】：郑州大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R737.9;R392

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