定義新型前列腺癌致癌及轉移驅動因素

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姓名	荷娜(Nurul Hayati) 查詢紙本館藏	畢業系所	生命科學系
論文名稱	定義新型前列腺癌致癌及轉移驅動因素 (Define Novel Prostate Cancer Oncogenic and Metastasis Driver)
檔案	[Endnote RIS 格式] [Bibtex 格式] [相關文章] [文章引用] [完整記錄] [館藏目錄] 至系統瀏覽論文 (2028-6-25以後開放)
摘要(中)	前列腺癌（PCa）仍然是發病和死亡的一個重要原因，病患帶有癌轉移腫瘤與較差的存活率有關。我們研究的目標是確定新的治療策略並改善癌轉移患者的預後。採用多方面的方法，結合臨床基因體分析、in vitro 和in vivo 實驗，發現前列腺癌（PCa）中新的致癌驅動因素和轉移促進因素，我們專注於一個精氨酸 N-降解決定子途徑相關的基因，稱為WDYHV1 (NTAQ1)。我們透過慢病毒轉染在LNCaP 前列腺癌細胞株過度表現該基因，證明了該基因的致癌功能，例如刺激增殖、集落形成、3D 侵襲和跨孔遷移。並且，我們發現WDYHV1 (NTAQ1)促進在NSG 免疫缺陷小鼠上的異質腫瘤生長與癌轉移。轉錄體分析顯示有顯著差異的585 個基因表現，透過路徑分析表明，上調基因參與上皮間質轉化 (EMT) 和透過NF-κB 進行的TNF-α 訊號傳導。這些發現凸顯了 WDYHV1 可作為開發減輕轉移性前列腺癌的標靶療法的有前途的候選基因。我未來的研究將專注於透過驗證轉錄體分析的發現和與尋找精氨酸 N-降解決定子調節相關的 WDYHV 直接受質來闡明潛在致癌機制。
摘要(英)	Prostate cancer (PCa) remains a significant cause of morbidity and mortality, and the presence of metastasis is linked to poor survival rates. The goal of our study is to identify novel therapeutic strategies and improve outcomes for patients with metastasis. A multifaceted approach was employed combining clinical genomic analysis, in vitro assays and in vivo assays to uncover novel oncogenic drivers and metastasis-promoting factors in prostate cancer (PCa) focusing on a specific gene associated with the arginine N-degron pathway, called WDYHV1 (NTAQ1). Via overexpression of WDYHV1(NTAQ1) in LNCaP prostate cancer cell line using lentiviral transduction, we demonstrated its oncogenic functions including stimulation of proliferation, colony formation, 3D invasion, and transwell migration ability. Furthermore, we found that WDYHV1(NTAQ1) promotes in vivo xenograft tumor growth and metastasis in NSG immunodeficient mice utilizing the luciferase / dTomato expression as the reporter. Transcriptome profiling uncovered that 585 genes upregulated dramatically and pathway analysis of these up-regulated genes indicated the involvement of epithelial-mesenchymal transition (EMT) and TNF-alpha signaling via NF-κB. These findings highlight the WDYHV1 gene as a promising candidate for developing targeted therapies to mitigate metastatic prostate cancer. My future studies will focus on elucidating the underlying oncogenic mechanism from transcriptome analysis and looking for potential direct WDYHV substracts associated with arginine N-degron regulation.
關鍵字(中)	★ 前列腺癌 ★ 轉移 ★ N-降解決定子途徑	關鍵字(英)	★ Prostate Cancer ★ metastasis ★ N-Degron pathway
論文目次	中文摘要 .................................................................................................................................... v Abstract ...................................................................................................................................... vi Acknowledgments ................................................................................................................... viii Table of Contents ...................................................................................................................... ix List of Figures ............................................................................................................................ xi Explanation of Symbols and Abbreviations ............................................................................. xii Chapter I. Introduction ............................................................................................................... 1 1-1 Epidemiology of Prostate Cancer (PCa) ................................................................. 1 1-2 Prostate Cancer Therapies ...................................................................................... 2 1-3 Molecular Biology of Prostate Cancer ................................................................... 4 1-4 N-terminal Glutamine Amidohydrolase (NTAQ) .................................................. 4 1-4-1 Protein Degradation ................................................................................................ 4 1-4-2 N-end Rule Pathways ............................................................................................. 6 1-4-3 WDYHV1: From PTM to Potential Cancers Implications ..................................... 7 1-5 Research Objectives ............................................................................................... 8 Chapter II. Materials and Methods ........................................................................................... 10 2-1 Generation of pHIV-dTomato-NSF-WDYHV1 ................................................... 10 2-1-1 Vector Design ....................................................................................................... 10 2-1-2 Construction of pHIV-dTomato-Luciferase Vector ............................................. 11 2-1-3 cDNA Synthesis ................................................................................................... 11 2-1-4 Polymerase Chain Reaction (PCR) ...................................................................... 12 2-1-5 Transfection and generation of stable cell lines ................................................... 13 2-2 Cell Culture .......................................................................................................... 14 2-3 Western Blotting Analysis and Antibodies .......................................................... 14 2-4 Cancer Cell Proliferation ...................................................................................... 15 2-5 Colony Formation ................................................................................................. 15 2-6 Cancer Cell Migration .......................................................................................... 15 2-7 Cancer Cell Invasion ............................................................................................ 16 2-8 Xenograft Experiment in NSG Mice .................................................................... 16 2-9 Experimental Metastasis Assays .......................................................................... 16 2-10 Transcriptome Analysis ........................................................................................ 17 x 2-10-1 RNA Extraction .................................................................................................... 17 2-10-2 Microarrays Analysis ............................................................................................ 17 2-11 Statistical Analysis ............................................................................................... 18 Chapter III. Results ................................................................................................................... 19 3-1 Identification of candidate gene for prostate cancer metastasis ........................... 19 3-2 WDYHV1 expression in prostate cancer cells ..................................................... 20 3-3 Western blot analysis of overexpressing WDYHV1 in LNCaP ........................... 20 3-4 WDYHV1 overexpression increased a proliferation rate in LNCaP .................... 21 3-5 WDYHV1 overexpression increases colony number and size ............................. 22 3-6 WDYHV1 overexpression promotes prostate cancer cell migration ................... 23 3-7 WDYHV1 overexpression promotes cancer cell invasion ................................... 23 3-8 WDYHV1 promotes tumor growth in-vivo .......................................................... 24 3-9 WDYHV1 promotes the metastasis ability .......................................................... 25 3-10 Transciptome analysis of WDYHV1 overexpression .......................................... 27 Chapter IV. Discussion ............................................................................................................. 29 4-1 General Discussion ............................................................................................... 29 4-2 Limitations and future works ................................................................................ 29 4-1-1 Gene knockdown of WDYHV1 ........................................................................... 29 4-1-2 Validating the mechanism based on transcriptome profiling ............................... 30 4-1-3 N-end Rule Degradation Pathway ........................................................................ 30 4-1-4 Potential Downstream Target ............................................................................... 31 Chapter V. Conclusion ............................................................................................................. 32 Bibliography ............................................................................................................................. 33 Appendix A. ............................................................................................................................. 41 Appendix B. .............................................................................................................................. 45
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指導教授	許恩旗范世榮(En-Chi Hsu Shih-Jung Fan)	審核日期	2024-7-26
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