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http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/22691完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 阮麗蓉(Li-Jung Juan) | |
| dc.contributor.author | Chung-Fan Lee | en |
| dc.contributor.author | 李中帆 | zh_TW |
| dc.date.accessioned | 2021-06-08T04:24:47Z | - |
| dc.date.copyright | 2010-09-09 | |
| dc.date.issued | 2010 | |
| dc.date.submitted | 2010-06-17 | |
| dc.identifier.citation | 1. Law, J.A., and Jacobsen, S.E. Establishing, maintaining and modifying DNA methylation patterns in plants and animals. Nat Rev Genet 11:204-220.
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/22691 | - |
| dc.description.abstract | 抑癌基因啟動子的高度甲基化抑制抑癌基因轉錄於腫瘤形成過程扮演重要角色，然而主導基因甲基化的甲基轉移酶活性調控及其結合至目標基因等機制未明。我們發現甲基轉移酶DNA methyltransferase 1 (DNMT1)可與負責催化蛋白質alpha-及epsilon-乙醯化反應的人類蛋白hNaa10p/hARD1產生交互作用。多項證據明確指出hNaa10p/hARD1可透過DNMT1使細胞癌化。我們發現hNaa10p的過量表現與肺癌患者的預後不佳具有正相關性。與此現象相符的是，當細胞大量表現外生性hNaa10p時，細胞出現轉型現象(cell transformation)。利用小片段干擾RNA減少肺癌細胞hNaa10p基因表現，可使癌細胞增殖能力受損，並減少其於小鼠體內形成腫瘤能力。hNaa10p的致癌能力主要與DNMT1的結合相關，而非其造成DNMT1乙醯化所致。我們更發現，hNaa10p可促進DNMT1與其受質DNA結合，並藉此增加DNMT1活性。細胞實驗亦證實hNaa10p可吸引DNMT1與抑癌基因(例如: E-cadherin) 啟動子結合，增加E-cadherin啟動子甲基化，並降低其基因表現。這些現象完全依賴hNaa10p與DNMT1結合。我們更進一步指出，E-cadherin並非唯一受影響基因。根據上述觀察，我們不僅確認hNaa10p為致癌基因，並釐清hNaa10p乃藉由調控DNMT1的功能促進腫瘤形成。 | zh_TW |
| dc.description.abstract | Hypermethylation-mediated tumor suppressor gene silencing plays a crucial role in tumorigenesis. Understanding its underlying mechanism is essential for cancer treatment. Previous studies on the hNaa10p/hARD1 (human arrest-defective 1), a human protein capable of catalyzing both N-alpha- and epsilon-acetylation, have generated conflicting results with regard to its role in tumorigenesis. Here we provide multiple lines of evidence supporting its oncogenic function. We show that hNaa10p overexpression correlates with poor survival of human lung cancer patients. Consistently, enforced expression of hNaa10p is sufficient to cause cellular transformation and siRNA-mediated depletion of hNaa10p impairs cancer cell proliferation in colony assays and xenograft studies. The oncogenic potential of hNaa10p seems to depend on its interaction with the DNA methyltransferase DNMT1, but not the hNaa10p-mediated DNMT1 acetylation. Mechanistically, hNaa10p positively regulates the enzymatic activity of DNMT1 by facilitating its binding to DNA in vitro and its recruitment to promoters of tumor suppressor genes, such as E-cadherin, in vivo. Importantly, interaction between hNaa10p and DNMT1 is required for E-cadherin silencing through promoter CpG methylation and E-cadherin repression contributes to the hNaa10p-mediated oncogenic property. Together our studies not only establish hNaa10p as an oncoprotein, but also reveal that hNaa10p contributes to oncogenesis through modulation of DNMT1 function. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-08T04:24:47Z (GMT). No. of bitstreams: 1 ntu-99-D93448004-1.pdf: 1907426 bytes, checksum: 3835eb44d78dee8c2188629185573b2d (MD5) Previous issue date: 2010 | en |
| dc.description.tableofcontents | Contents I
List of Figures IV List of Tables VI 中文摘要 1 Abstract 2 Chapter I: Introduction 3 1. DNA methyltransferase (DNMT) family 4 1.1 DNA methylation, the epigenetic modification catalyzed by DNMT family 4 1.2 Aberrant DNA methylation in cancer 5 1.3 Regulation of DNMTs 6 1.4 Functional domains and interacting proteins of DNMT1 8 2. human Naa10p/human arrest-defective 1 11 2.1 N-α-acetylation, the conserved biological functions of Naa10p 11 2.2 Naa10p, a conserved protein acetyltransferase throughout eukaryotic kindom 12 2.3 Biological functions of mammalian Naa10p 12 2.4 Dysregulation of Naa10p in cancer 13 Specific Aims 15 Chapter II: Materials and Methods 16 Lung cancer patients, clinical sample preparation, real-time PCR and IHC assay 17 Cell culture and transfection 18 Plasmids and siRNAs 18 Antibodies 19 In vivo acetylation assay 20 In vitro auto-acetylation assay 20 Generation of stable clones by lentivirus infection, cell proliferation, soft agar and NOD-SCID mice tumorigenicity assays 20 IP, Western, and luciferase assay 21 GST pull-down and EMSA assays 21 DNA methyltransferase assay (DNMT assay) 22 ChIP and cDNA analysis 23 Bisulfite sequencing 23 Statistics 24 Chapter III: Results 25 1. hNaa10p overexpression is correlated to the poor prognosis in human lung cancer tissues 26 2. hNaa10p contributes to clonogenesis in lung cancer cells 26 3. hNaa10p associates with DNMT1 27 4. hNaa10p association with DNMT1 is crucial for its oncogenic potential 28 5. hNaa10p maintains and stimulates the DNMT1 methyltransferase activity 29 6. hNaa10p enhances the DNA-binding ability of DNMT1 both in vitro and in vivo 30 7. hNaa10p regulates the methylation on E-cadherin promoter 31 8. hNaa10p-mediated repression of E-cadherin and LATS2 in DNMT1-dependent manner contributes to hNaa10p oncogenic ability 32 Chapter IV: Summary and Discussion 35 Figures 40 Tables 59 References 64 | |
| dc.language.iso | en | |
| dc.subject | 癌症 | zh_TW |
| dc.subject | DNA甲基轉移酶 | zh_TW |
| dc.subject | 乙醯基轉移酶 | zh_TW |
| dc.subject | hNaa10p/hARD1 | zh_TW |
| dc.subject | 表遺傳調控 | zh_TW |
| dc.subject | 高度甲基化抑癌基因 | zh_TW |
| dc.subject | cancer | en |
| dc.subject | hNaa10p/hARD1 (human arrest-defective 1) | en |
| dc.subject | DNA methyltransferase | en |
| dc.subject | epigenetic regulation | en |
| dc.subject | hypermethylated tumor suppressor gene | en |
| dc.title | 乙醯基轉移酶hNaa10p/hARD1協助DNA甲基轉移酶DNMT1抑制抑癌基因活化之機制探討 | zh_TW |
| dc.title | hNaa10p/Human arrest-defective 1 contributes to tumorigenesis by facilitating DNMT1-mediated tumor suppressor gene silencing | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 98-2 | |
| dc.description.degree | 博士 | |
| dc.contributor.coadvisor | 吳成文(Cheng-Wen Wu) | |
| dc.contributor.oralexamcommittee | 李芳仁(Fang-Jen Lee),王憶卿(Yi-Ching Wang),施修明(Hsiu-Ming Shih) | |
| dc.subject.keyword | DNA甲基轉移酶,乙醯基轉移酶,hNaa10p/hARD1,表遺傳調控,高度甲基化抑癌基因,癌症, | zh_TW |
| dc.subject.keyword | DNA methyltransferase,hNaa10p/hARD1 (human arrest-defective 1),epigenetic regulation,hypermethylated tumor suppressor gene,cancer, | en |
| dc.relation.page | 79 | |
| dc.rights.note | 未授權 | |
| dc.date.accepted | 2010-06-17 | |
| dc.contributor.author-college | 醫學院 | zh_TW |
| dc.contributor.author-dept | 分子醫學研究所 | zh_TW |
| 顯示於系所單位： | 分子醫學研究所 | |
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