本文選題：ILF3 + 雌激素受體��； 參考：《浙江理工大學》2012年碩士論文

【摘要】：近年來，mRNA非翻譯區(qū)的功能研究成為生物學研究的熱點之一。在mRNA的3’非翻譯區(qū)有許多調(diào)控元件，而這些調(diào)控元件能與蛋白質或蛋白質復合物結合，調(diào)控mRNA的翻譯， mRNA的穩(wěn)定性，以及mRNA亞細胞定位等。 ERβ在乳腺癌、前列腺癌、結腸癌、和卵巢癌的發(fā)生、發(fā)展過程中有調(diào)控作用。本文利用RNA沉降技術，研究ERβ3’UTR mRNA相關調(diào)控機制，得到與其特異性結合的一種重要蛋白質：ILF3。 ILF3能與富含AU序列的mRNAs特異性結合，調(diào)控基因表達。對其功能的研究對深入了解雌激素信號通路的調(diào)控以及作用機制有重要意義。 ILF3是白介素增強結合因子3，是雙鏈RNA結合蛋白中的一員，它位于人類19號染色體上，編碼一個894個氨基酸的蛋白。ILF3蛋白參與RNA轉錄、RNA剪切、RNA編輯、RNA出核轉運、RNA的亞細胞定位等，涉及細胞發(fā)育，細胞周期調(diào)控和病毒感染等多重細胞功能。 本文通過Western Blot，， EMSA等實驗證明了ILF3能特異的結合雌激素受體（ERβ）的3’UTR。雌激素受體可介導雌激素實現(xiàn)其生物學功能， ILF3蛋白與ERβ3’UTR結合，因此ILF3蛋白可能參與了雌激素信號通路的調(diào)控。本文以雌二醇誘導乳腺癌細胞MCF7，發(fā)現(xiàn)了ILF3能夠被雌激素誘導表達。為探討ILF3蛋白與雌激素受體功能關系，通過構建過表達和低表達ILF3蛋白的細胞株，Western blot檢測ILF3蛋白變化對ERβ的影響，并通過MTT實驗檢測ILF3蛋白表達量下調(diào)對MCF7細胞活力的影響。實驗結果說明：雌激素可誘導ILF3蛋白表達量上調(diào)；ILF3過表達導致ERβ表達量增加而低表達則ERβ表達量下降。ILF3在蛋白水平上對ERβ起正向調(diào)控。MTT實驗結果表明ILF3低表達對MCF7細胞生長活力沒有明顯的影響。 構建低表達ERβ蛋白的細胞株，實驗結果表明ERβ蛋白表達量下調(diào)，ILF3蛋白含量也下降，ERβ與ILF3起協(xié)同作用。為揭示ILF3的功能及其在ERβ的調(diào)控中的作用機制提供理論依據(jù)，并為更深入地研究雌激素信號通路奠定了基礎。
[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.
【學位授予單位】：浙江理工大學
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R341

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