RNA结合蛋白ILF3在雌激素信号通路中的功能研究

发布时间：2018-04-22 11:37

本文选题：ILF3 + 雌激素受体　；参考：《浙江理工大学》2012年硕士论文

【摘要】：近年来，mRNA非翻译区的功能研究成为生物学研究的热点之一。在mRNA的3’非翻译区有许多调控元件，而这些调控元件能与蛋白质或蛋白质复合物结合，调控mRNA的翻译， mRNA的稳定性，以及mRNA亚细胞定位等。 ERβ在乳腺癌、前列腺癌、结肠癌、和卵巢癌的发生、发展过程中有调控作用。本文利用RNA沉降技术，研究ERβ3’UTR mRNA相关调控机制，得到与其特异性结合的一种重要蛋白质：ILF3。 ILF3能与富含AU序列的mRNAs特异性结合，调控基因表达。对其功能的研究对深入了解雌激素信号通路的调控以及作用机制有重要意义。 ILF3是白介素增强结合因子3，是双链RNA结合蛋白中的一员，它位于人类19号染色体上，编码一个894个氨基酸的蛋白。ILF3蛋白参与RNA转录、RNA剪切、RNA编辑、RNA出核转运、RNA的亚细胞定位等，涉及细胞发育，细胞周期调控和病毒感染等多重细胞功能。本文通过Western Blot，， EMSA等实验证明了ILF3能特异的结合雌激素受体（ERβ）的3’UTR。雌激素受体可介导雌激素实现其生物学功能， ILF3蛋白与ERβ3’UTR结合，因此ILF3蛋白可能参与了雌激素信号通路的调控。本文以雌二醇诱导乳腺癌细胞MCF7，发现了ILF3能够被雌激素诱导表达。为探讨ILF3蛋白与雌激素受体功能关系，通过构建过表达和低表达ILF3蛋白的细胞株，Western blot检测ILF3蛋白变化对ERβ的影响，并通过MTT实验检测ILF3蛋白表达量下调对MCF7细胞活力的影响。实验结果说明：雌激素可诱导ILF3蛋白表达量上调；ILF3过表达导致ERβ表达量增加而低表达则ERβ表达量下降。ILF3在蛋白水平上对ERβ起正向调控。MTT实验结果表明ILF3低表达对MCF7细胞生长活力没有明显的影响。构建低表达ERβ蛋白的细胞株，实验结果表明ERβ蛋白表达量下调，ILF3蛋白含量也下降，ERβ与ILF3起协同作用。为揭示ILF3的功能及其在ERβ的调控中的作用机制提供理论依据，并为更深入地研究雌激素信号通路奠定了基础。
[Abstract]:In recent years, the functional study of untranslated region of mRNAs has become one of the hotspots in biological research. There are many regulatory elements in the 3'untranslated region of mRNA, which can bind to protein or protein complex, regulate the translation of mRNA, the stability of mRNA, and the subcellular localization of mRNA. ER-尾 plays a regulatory role in the development and progression of breast, prostate, colon, and ovarian cancers. In this paper, we studied the regulation mechanism of ER 尾 3'UTR mRNA by RNA sedimentation technique, and got an important protein,: ILF3, which specifically binds to ER 尾 3'UTR. ILF3 can specifically bind to mRNAs rich in AU sequences and regulate gene expression. The study of its function is of great significance in understanding the regulation and mechanism of estrogen signaling pathway. ILF3 is a member of double-stranded RNA binding protein. It is located on human chromosome 19 and encodes an 894 amino acid protein. ILF3 protein is involved in the subcellular localization of RNA transporter. Multiple cell functions are involved in cell development, cell cycle regulation and viral infection. In this paper, Western blot, EMSA and other experiments have proved that ILF3 can specifically bind to estrogen receptor ER 尾. Estrogen receptor can mediate estrogen to realize its biological function, and ILF3 protein binds to ER 尾 3'UTR, so ILF3 protein may be involved in the regulation of estrogen signaling pathway. In this paper, we found that ILF3 can be induced by estrogen in breast cancer cell line MCF7 induced by estradiol. In order to study the relationship between ILF3 protein and estrogen receptor function, we constructed a cell line with overexpression and low expression of ILF3 protein to detect the effect of ILF3 protein on ER 尾 by Western blot, and to detect the effect of down-regulation of ILF3 protein expression on the viability of MCF7 cells by MTT assay. The results showed that estrogen could up-regulate the expression of ILF3 protein and increase the expression of ER 尾. The results showed that the low expression of ER 尾 and the positive regulation of ILF3 on ER 尾 at the protein level showed that the low expression of ILF3 had a positive effect on the expression of ER 尾. The growth activity of MCF7 cells was not significantly affected. A cell line with low expression of ER 尾 protein was constructed. The results showed that the down-regulation of ER 尾 protein expression also decreased the content of ILF3 protein. ER 尾 and ILF3 played a synergistic role. It provides a theoretical basis for revealing the function of ILF3 and its mechanism in the regulation of ER 尾, and lays a foundation for further study of estrogen signaling pathway.
【学位授予单位】：浙江理工大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R341

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