English  |  正體中文  |  简体中文  |  全文筆數/總筆數 : 45073/58249 (77%)
造訪人次 : 2398385      線上人數 : 151
RC Version 7.0 © Powered By DSPACE, MIT. Enhanced by NTU Library IR team.
搜尋範圍 查詢小技巧：
  • 您可在西文檢索詞彙前後加上"雙引號"，以獲取較精準的檢索結果
  • 若欲以作者姓名搜尋，建議至進階搜尋限定作者欄位，可獲得較完整資料
    •  進階搜尋


    請使用永久網址來引用或連結此文件: http://libir.tmu.edu.tw/handle/987654321/51019


    題名: D-antroquinonol為一新穎之DNA甲基轉移酶抑制劑，可抑制DNA甲基轉移酶-1活性且對於人類乳癌細胞具有抗癌效果
    A Novel DNA Methyltransferase Inhibitor, D-antroquinonol, Inhibits The DNMT-1 Activity and Induces Anticancer Effects on Human Breast Cancer Cells
    作者: 王昇超
    Wang, Sheng-Chao
    關鍵詞: DNA甲基轉移酶;乳癌;抗癌;DNA Methyltransferase;Breast Cancer;Anticancer
    日期: 2013-05-28
    上傳時間: 2018-10-05 10:21:57 (UTC+8)
    摘要: DNA methylation是一種epigenetic modification機制，可不造成DNA鹼基序列異動而調節gene表現。DNA methylation於癌症中Tumor suppressor genes (TSGs) 之siliencing扮演著重要之關鍵。然而DNA methyltransferases (DNMTs) 是催化DNA methylation之酵素，並且過去文獻報導DNMTs protein表現量於多種癌症中為過度表現情形。此外，TSGs之promoter hypermethylation可透過抑制DNMTs而可逆地恢復為正常狀態，因此DNMT抑制劑 (DNMT inhibitor; DNMTi) 之開發是極具新藥開發潛力之領域。
    D-antroquinonol是一個自牛樟芝 (Antrodia camphorata) 純化萃取而得且過去從未被報導過之化合物。為了評估D-antroquinonol是否具有抑制DNA甲基轉移酶1 (DNA methyltransferase1; DNMT1) 之活性，並且透過降低TSGs相關之CpG islands之promoter hypermethylation情形以重新活化TSGs表現以抑制乳癌細胞之生長與轉移。利用核磁共振分析儀 (Nuclear magnetic resonance ; NMR) 鑑定D-antroquinonol 結構為3-demethoxyl antroquinonol. 利用DNMT1酵素活性分析、分子模型與對接分析 (molecular modeling and docking)、the Illumina Methylation 450K array-based assay、pyrosequencing、即時定量RT-PCR及Western blotting分析此化合物對於乳癌細胞(MDA-MB-231) 之特異性及抗癌效果。DNMT1酵素活性分析具有隨著投予之化合物劑量增加而有酵素活性抑制之效果且此分析在DNMT3B酵素活性分析未被檢出相同結果，推測此化合物對於DNMT1酵素具有特異性。而molecular docking之結果顯示D-antroquinonol可binding於DNMT1之催化位。D-antroquinonol 可降低乳癌細胞 (MDA-MB-231) 內多種TSGs之甲基化程度，包括FANCC和CACNA1A基因，Western blotting及real time RT-PCR analysis 顯示D-antroquinonol 可誘導增加FANCC, CACNA1A之mRNA及蛋白質表現量。而SRB分析結果顯示D-antroquinonol可誘導抑制乳癌細胞 (MCF-7、T-47D及MDA-MB- 231) 之生長且不影響正常細胞生長 (IMR-90及MCF-10A)。The wound-healing assay及transwell assays 結果顯示D-antroquinonol可誘導抑制乳癌細胞 (MDA-MB- 231) 之爬行能力。
    總結以上，我們鑑定了一個過去未被發表過之DNMT1抑制劑－D-antroquinonol，可誘導DNA 之demethylation、恢復TSGs表現並抑制癌細胞生長及轉移。

    DNA methylation is an epigenetic modification that modulates gene expression without altering the DNA base sequence. It plays a crucial role in cancer by silencing tumor suppressor genes (TSGs). The DNA methyltransferases (DNMTs) are the enzymes that catalyze DNA methylation. DNMTs proteins have been reported that overexpress in various cancers. In addition, promoter hypermethylation of tumor suppressor genes is reversible by inhibition of DNMTs. Therefore, there is a need to development of DNMT inhibitors.
    D-antroquinonol is a novel compound isolated from Antrodia camphorata. To investigate whether the D-antroquinonol is potent to inhibit the DNMTs’ enzyme activity and reactivate the TSGs by downregulating the promoter hypermethylation of TSG-associated CpG islands, and result in suppression of breast cancer cells growth and migration. D-antroquinonol (3-demethoxyl antroquinonol) was identified by nuclear magnetic resonance. The drug targeting and anticancer effects of D-antroquinonol to breast cancer cells (MDA-MB-231) were determined by the DNMT1 enzyme activity assay, molecular modeling and docking, the Illumina Methylation 450K array-based assay, pyrosequencing, real-time RT-PCR, Western blotting. Cell cytotoxicity, growth inhibition and migration ability were also analyzed. The D-antroquinonol inhibited DNMT1 in a dose-dependent manner but not DNMT3B. We speculated that D-antroquinonol can inhibit DNMT1 activity specifically. Molecular docking and enzyme activity data revealed
    that D-antroquinonol may bind at catalytic domain of DNMT1. The D-antroquinonol decreased the methylation level of multiple TSGs, including the FANCC and CACNA1A genes, in MDA-MB-231 breast cancer cells. Western blot and RT-PCR analysis showed that D-antroquinonol increased FANCC, CACNA1A mRNA and protein expression levels. The SRB assay result indicated that D-antroquinonol inhibits the growth of breast cancer cells (MCF-7, T-47D and MDA-MB-231) but not normal cells (IMR-90 and MCF-10A). The results of wound-healing assay and transwell assays suggested that D-antroquinonol inhibits the migration of MDA-MB-231 breast cancer cells.
    In conclusion, we identified a novel DNMT inhibitor, D-antroquinonol, which induced DNA demethylation, recovered the expression of multiple tumor suppressor genes, and inhibited cancer cell growth and metastasis.
    描述: 碩士
    委員-阮麗蓉
    委員-劉興璟
    指導教授-林若凱
    資料類型: thesis
    顯示於類別:[生藥學研究所] 博碩士論文

    文件中的檔案:

    檔案 描述 大小格式瀏覽次數
    index.html0KbHTML366檢視/開啟


    檢視Licence

    在TMUIR中所有的資料項目都受到原著作權保護.

    TAIR相關文章

    著作權聲明 Copyright Notice

    • 本平台之數位內容為臺北醫學大學所收錄之機構典藏，包含體系內各式學術著作及學術產出。秉持開放取用的精神，提供使用者進行資料檢索、下載與取用，惟仍請適度、合理地於合法範圍內使用本平台之內容，以尊重著作權人之權益。商業上之利用，請先取得著作權人之授權。

      The digital content on this platform is part of the Taipei Medical University Institutional Repository, featuring various academic works and outputs from the institution. It offers free access to academic research and public education for non-commercial use. Please use the content appropriately and within legal boundaries to respect copyright owners' rights. For commercial use, please obtain prior authorization from the copyright owner.

    • 瀏覽或使用本平台，視同使用者已完全接受並瞭解聲明中所有規範、中華民國相關法規、一切國際網路規定及使用慣例，並不得為任何不法目的使用TMUIR。

      By utilising the platform, users are deemed to have fully accepted and understood all the regulations set out in the statement, relevant laws of the Republic of China, all international internet regulations, and usage conventions. Furthermore, users must not use TMUIR for any illegal purposes.

    • 本平台盡力防止侵害著作權人之權益。若發現本平台之數位內容有侵害著作權人權益情事者，煩請權利人通知本平台維護人員([email protected])，將立即採取移除該數位著作等補救措施。

      TMUIR is made to protect the interests of copyright owners. If you believe that any material on the website infringes copyright, please contact our staff([email protected]). We will remove the work from the repository.

    Back to Top
    DSpace Software Copyright © 2002-2004  MIT &  Hewlett-Packard  /   Enhanced by   NTU Library IR team Copyright ©   - 回饋