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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 詹世鵬(Shih-Peng Chan) | |
| dc.contributor.author | Yi-Jun Wu | en |
| dc.contributor.author | 吳翊均 | zh_TW |
| dc.date.accessioned | 2021-06-17T01:16:30Z | - |
| dc.date.available | 2022-09-14 | |
| dc.date.copyright | 2017-09-14 | |
| dc.date.issued | 2017 | |
| dc.date.submitted | 2017-08-14 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/66989 | - |
| dc.description.abstract | 異質性核核醣核酸蛋白(Heterogeneous nuclear ribonucleoproteins, hnRNPs) 是一個至少包含了二十種以上核醣核酸結合蛋白家族，大多數的異質性核核醣核酸蛋白主要表現在細胞核當中，由於其都具有核糖核酸結合區域 (RNA-binding domains)，因此異質性核核醣核酸蛋白在功能上被認為會參與許多核醣核酸的生合成與代謝路徑，包括在細胞核內基因的轉錄、訊息核醣核酸的剪接與運送等等。異質性核核醣核酸蛋白能隨轉運至細胞核中，參與調控轉譯形成蛋白質之過程以及訊息核糖核酸的穩定性。除此之外，異質性核核醣核酸蛋白也會參與非編碼核糖核酸(Non-coding RNAs)，比如微小核醣核酸 (microRNAs, miRNAs)的生合成與功能。我們於先前的研究當中，發現線蟲異質性核核醣核酸蛋白HRP-2能夠藉由結合在lin-41訊息核醣核酸的三端不轉譯區，來干擾let-7微小核醣核酸對lin-41誘發之基因沉默。HRP-2在人類當中的同源蛋白質包含異質核醣核酸蛋白Q和R (hnRNP Q/R)，我們進一步發現hnRNP Q有類似HRP-2在線蟲當中的功能。HNRNPQ的基因敲落，會造成TRIM71蛋白質表現量的大幅下降，我們證明了此現象是由於hnRNP Q能夠藉由結合在TRIM71訊息核醣核酸的三端不轉譯區，干擾let-7誘發的基因沉默現象 (論文尚未發表)。lin-41 (lineage variant 41)/TRIM71 (tripartite motif 71) 基因編有E3泛素連接酶的密碼訊息，其訊息核糖核酸受到抑癌微小核醣核酸let-7調控是為廣為人知。LIN41/TRIM71蛋白質亦具有結合核醣核酸的能力，在先前研究被報導和幹細胞的分化與複製以及癌細胞的分裂複製相關。在這篇研究當中，我們想研究線蟲HRP-2在人類中同源蛋白hnRNP R，是否同樣會調控TRIM71基因的表現。我們利用TRIM71蛋白質相對高表現量的肝癌細胞株Huh7作為實驗材料，將HNRNPR基因敲落，發現TRIM71的蛋白質及mRNA的表現都有顯著性的下降。同樣的趨勢也在另一肝癌細胞株HepG2中被觀察到。除此之外我們發現在缺乏hnRNP R 蛋白質的情況之下，TRIM71 mRNA的穩定性會降低。hnRNP R蛋白質調控TRIM71基因表現的機制以及其與let-7之間是否存在類似hnRNP Q阻斷let-7對於 TRIM71負向功能關聯性，仍有待未來進一步探討。 | zh_TW |
| dc.description.provenance | Made available in DSpace on 2021-06-17T01:16:30Z (GMT). No. of bitstreams: 1 ntu-106-R04445117-1.pdf: 1915839 bytes, checksum: 8d4ddf70766260e5544d22789d9f8b8f (MD5) Previous issue date: 2017 | en |
| dc.description.tableofcontents | 口試委員會審定書 Ⅰ
誌謝 Ⅱ 中文摘要 Ⅲ ABSTRACT Ⅳ CONTENTS 1 Chapter 1 Introduction 4 1.1 heterogeneous nuclear ribonucleoproteins (hnRNPs) 4 1.1.1 The hnRNPs family and their structural features 4 1.1.2 Functional diversity of hnRNPs 5 1.1.3 Link of hnRNPs to diseases 7 1.1.4 hnRNP R 8 1.2 Lin-41/TRIM71 8 1.3 Project aims 9 Chapter 2 Materials and Methods 11 2.1 Cell culture and Transfection 11 2.2 Lentivirus packging and infection 13 2.3 Cell lysate preperationand protein quantitation 14 2.4 Nuclear / Cytoplasmic fractionation 15 2.5 Immunoblotting 16 2.6 Plasmids 17 2.6.1 Vectors 17 2.6.2 Constructs 18 2.7 RNA isolation 19 2.8 Reverse transcription 20 2.8.1 Random priming-based reverse transcription 20 2.8.2 Poly-A tail-based reverse transcription 21 2.9 Quantitative real time polymerase chain reaction 22 2.10 mRNA stability assay 24 Chapter 3 Results 25 3.1 hnRNP R knockdown leads to a decrease in TRIM71 protein levels 25 3.2 hnRNP R knockdown leads to a decrease in TRIM71 mRNA levels 26 3.3 The TRIM71 mRNA stability is influenced by hnRNP R knockdown 26 3.4 Exogenous hnRNP R expression partially restores TRIM71 expression in hnRNP R knocked down Huh7 cells 27 Chapter 4 Discussion 29 4.1 Expression of hnRNP R in hnRNP R depleted Huh7 cells partially restores the reduced TRIM71 protein levels 29 4.2 LIN28B expression may be influenced by hnRNP R knockdown in HepG2 but not Huh7 cells 29 4.3 hnRNP R knockdown increases the protein levels of hnRNP Q2/Q3 30 Chapter 5 Figures 32 Figure 1. hnRNP R knockdown leads to a decrease in TRIM71 protein levels in Huh7 cells. 32 Figure 2. hnRNP R knockdown leads to a decreased in TRIM71 protein levels in HepG2 cells 33 Figure 3. hnRNP R knockdown by siRNAs transfection did not affect the protein levels of TRIM71 in Huh7 cells 35 Figure 4. hnRNP R knockdown significantly decreased TRIM71 mRNA levels in both Huh7 and HepG2 cells 36 Figure 5. The stability of TRIM71 mRNA is influenced by hnRNP R knockdown in Huh7 cells 38 Figure 6. Expression of hnRNP R fusion protein in hnRNP R knockdown Huh7 cells 40 Chapter 6 References 42 Chapter 7 Appendix 48 Appendix 1 . 48 Appendix 2 . 49 Appendix 3 . 51 | |
| dc.language.iso | zh-TW | |
| dc.subject | 異質性核核醣核酸蛋白Q/R | zh_TW |
| dc.subject | let-7微小核糖核酸 | zh_TW |
| dc.subject | lin-41/TRIM71 | zh_TW |
| dc.subject | HRP-2 | zh_TW |
| dc.subject | lin-41/TRIM71 | en |
| dc.subject | hnRNP Q/R | en |
| dc.subject | HRP-2 | en |
| dc.subject | let-7 miRNA | en |
| dc.title | 研究異質性核核醣核酸蛋白R對於TRIM71基因表現的調控機制 | zh_TW |
| dc.title | Study of the regulatory role for heterogeneous nuclear ribonucleoprotein R in TRIM71 expression | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 105-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 譚婉玉(Woan-Yuh Tarn),朱家瑩(Chia-Ying Chu) | |
| dc.subject.keyword | 異質性核核醣核酸蛋白Q/R,lin-41/TRIM71,HRP-2,let-7微小核糖核酸, | zh_TW |
| dc.subject.keyword | hnRNP Q/R,lin-41/TRIM71,HRP-2,let-7 miRNA, | en |
| dc.relation.page | 51 | |
| dc.identifier.doi | 10.6342/NTU201702840 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2017-08-14 | |
| dc.contributor.author-college | 醫學院 | zh_TW |
| dc.contributor.author-dept | 微生物學研究所 | zh_TW |
| 顯示於系所單位： | 微生物學科所 | |
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