去甲基化酶LSD1和JMJD1A在雄性激素调控的细胞中转录调控的分子机制的研究

发布时间：2018-04-18 22:09

  本文选题：组蛋白去甲基化 + LSD1　； 参考：《华东师范大学》2010年硕士论文

【摘要】：组蛋白N末端的各种转录后共价修饰可影响染色体的结构和调控基因的转录,组蛋白的甲基化修饰就是其中一种。胺基氧化酶家族的组蛋白去甲基化酶LSD1和JmjC家族的组蛋白去甲基化酶JMJD1A都参与雄性激素受体调控的转录激活过程,而且它们的异常表达与前列腺癌的发生发展相关,但对其分子机制的了解却很少。我们用慢病毒载体分别将去甲基化酶LSD1和JMJD1A两种蛋白的shRNA质粒转入前列腺癌LNCaP细胞中,得到稳定降低LSD1或JMJD1A表达的细胞系。然后我们通过用反转录聚合酶链反应实验和染色体免疫共沉淀实验发现,降低LSD1或JMJD1A的表达,都能在一定程度上抑制雄性激素调控的下游基因(例如PSA和NKX3.1)的表达,而且在雄性激素依赖的转录激活过程中,降低LSD1或JMJD1A的表达不影响染色体重塑的发生、组蛋白H3的乙酰化修饰水平和H3K4me2修饰水平的增高；同时对雄性激素受体与其靶基因上游的雄性激素受体应答元件的识别和结合、雄性激素受体对RNA聚合酶Ⅱ、以及共激活因子SRC家族(SRC-1, SRC-3)和CBP等的招募也没有很明显的影响。而且与以前的报道一致的是,降低JMJD1A和LSD1的表达使组蛋白H3K9me2修饰增高。更有趣的是,降低LSD1的表达可抑制了雄性激素受体对介质蛋白TRAP220在转录激活过程中的招募,而降低JMJD1A的表达却没有这样的现象。我们的结果表明同为组蛋白H3K9me/H3K9me2位的去甲基化酶JMJD1A和LSD1的调控机制存在一定差异,而且LSD1可能还通过控制对介质蛋白的招募而参与调控雄性激素诱导的转录激活过程。
[Abstract]:Various post-transcriptional modifications of histone N-terminal can affect the structure of chromosomes and the transcription of regulatory genes. Histone methylation is one of them.The histone demethylase (LSD1) of the amine oxidase family and the histone demethylase (JMJD1A) of the JmjC family are involved in the transcriptional activation of androgen receptor regulation, and their abnormal expression is related to the occurrence and development of prostate cancer.But little is known about its molecular mechanism.The shRNA plasmids of demethylase LSD1 and JMJD1A were transferred into LNCaP cells of prostate cancer with lentivirus vectors, respectively, to obtain a cell line that stably reduced the expression of LSD1 or JMJD1A.Then, by reverse transcriptase polymerase chain reaction and chromosome immunoprecipitation, we found that lowering the expression of LSD1 or JMJD1A could inhibit the expression of downstream genes (such as PSA and NKX3.1) regulated by androgen to some extent.In the course of androgen dependent transcriptional activation, the decrease of LSD1 or JMJD1A expression did not affect the occurrence of chromosome remodeling, and the acetylation modification level of histone H3 and the increase of H3K4me2 modification level were observed.At the same time, the recognition and binding of the male hormone receptor to the response element of the male hormone receptor upstream of the target gene was not obvious. The male hormone receptor had no significant effect on the recruitment of RNA polymerase 鈪，

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