請用此 Handle URI 來引用此文件:
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/62311
完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 林淑萍(Shwu-Bin Lin) | |
dc.contributor.author | Yun-Ju Lin | en |
dc.contributor.author | 林芸如 | zh_TW |
dc.date.accessioned | 2021-06-16T13:40:18Z | - |
dc.date.available | 2018-09-24 | |
dc.date.copyright | 2013-09-24 | |
dc.date.issued | 2013 | |
dc.date.submitted | 2013-07-15 | |
dc.identifier.citation | 1 Lin, Z. B. Cellular and molecular mechanisms of immuno-modulation by Ganoderma lucidum. Journal of Pharmacological Sciences 99, 144-153, (2005).
2 Xu, Z. T., Chen, X. U., Zhong, Z. F., Chen, L. D. & Wang, Y. T. Ganoderma lucidum Polysaccharides: Immunomodulation and Potential Anti-Tumor Activities. American Journal of Chinese Medicine 39, 15-27, (2011). 3 Russell, R. & Paterson, M. Ganoderma - A therapeutic fungal biofactory. Phytochemistry 67, 1985-2001, (2006). 4 Chen, N. H., Liu, J. W. & Zhong, J. J. Ganoderic acid T inhibits tumor invasion in vitro and in vivo through inhibition of MMP expression. Pharmacol Rep 62, 150-163, (2010). 5 Tang, W., Liu, J. W., Zhao, W. M., Wei, D. Z. & Zhong, J. J. Ganoderic acid T from Ganoderma lucidum mycelia induces mitochondria mediated apoptosis in lung cancer cells. Life Sci 80, 205-211, (2006). 6 賴曉萱. 靈芝酸T對肺癌細胞株之抗癌活性研究, Master thesis, 台灣大學, (2010). 7 潘宏韋. 靈芝萃取物T-612對非小細胞肺癌細胞株之抗癌活性研究, Master thesis, (2011). 8 蔡黛華. 靈芝三萜T-612透過誘導細胞自噬以促進Sorafenib於肝癌細胞株之效能, Master thesis, 台灣大學, (2012). 9 American Cancer Society breast cancer facts and figures http://www.cancer.org/research/ 10 Sandhu, R., Parker, J. S., Jones, W. D., Livasy, C. A. & Coleman, W. B. Microarray-Based Gene Expression Profiling for Molecular Classification of Breast Cancer and Identification of New Targets for Therapy. Labmedicine 41, 364-372, (2010). 11 Reis, J. S. & Pusztai, L. Breast Cancer 2 Gene expression profiling in breast cancer: classification, prognostication, and prediction. Lancet 378, 1812-1823, (2011). 12 Eroles, P., Bosch, A., Perez-Fidalgo, J. A. & Lluch, A. Molecular biology in breast cancer: Intrinsic subtypes and signaling pathways. Cancer Treatment Reviews 38, 698-707, (2012). 13 Ruder, A. M. et al. Estrogen and progesterone receptors in breast cancer patients. Epidemiologic characteristics and survival differences. Cancer 64, 196-202, (1989). 14 Del Mastro, L. et al. Trastuzumab as first-line therapy in HER2-positive metastatic breast cancer patients. Expert Rev Anticancer Ther 12, 1391-1405, (2012). 15 Nielsen, D. L., Kumler, I., Palshof, J. A. & Andersson, M. Efficacy of HER2-targeted therapy in metastatic breast cancer. Monoclonal antibodies and tyrosine kinase inhibitors. Breast 22, 1-12, (2013). 16 Rakha, E. A., Reis-Filho, J. S. & Ellis, I. O. Basal-like breast cancer: A critical review. Journal of Clinical Oncology 26, 2568-2581, (2008). 17 Deblois, G. & Giguere, V. Oestrogen-related receptors in breast cancer: control of cellular metabolism and beyond. Nat Rev Cancer 13, 27-36, (2013). 18 Zeng, Q. et al. Treating triple-negative breast cancer by a combination of rapamycin and cyclophosphamide: An in vivo bioluminescence imaging study. European Journal of Cancer 46, 1132-1143, (2010). 19 Schneider, B. P. et al. Triple-Negative Breast Cancer: Risk Factors to Potential Targets. Clinical Cancer Research 14, 8010-8018, (2008). 20 Dent, R. et al. Triple-negative breast cancer: Clinical features and patterns of recurrence. Clinical Cancer Research 13, 4429-4434, (2007). 21 Black, D. M. & Mittendorf, E. A. Landmark trials affecting the surgical management of invasive breast cancer. Surg Clin North Am 93, 501-518, (2013). 22 Barry, M. & Sacchini, V. Evaluating the role of intra-operative radiation therapy in the modern management of breast cancer. Surg Oncol 21, e159-163, (2012). 23 Breast Cancer Organization http://www.breastcancer.org 24 Wang, J. C. Cellular roles of DNA topoisomerases: A molecular perspective. Nature Reviews Molecular Cell Biology 3, 430-440, (2002). 25 Pommier, Y., Leo, E., Zhang, H. L. & Marchand, C. DNA Topoisomerases and Their Poisoning by Anticancer and Antibacterial Drugs. Chemistry & Biology 17, 421-433, (2010). 26 林亭妤. 天然物烯烷基對苯二酚[HQ17(3)]與硫醇基反應引發分子及細胞反應之研究, Doctor thesis, 台灣大學, (2013). 27 Fortune, J. M. & Osheroff, N. Topoisomerase II as a target for anticancer drugs: When enzymes stop being nice. Progress in Nucleic Acid Research and Molecular Biology, Vol 64 64, 221-253, (2000). 28 Li, T. K. & Liu, L. F. Tumor cell death induced by topoisomerase-targeting drugs. Annual Review of Pharmacology and Toxicology 41, 53-77, (2001). 29 Nitiss, J. L. Targeting DNA topoisomerase II in cancer chemotherapy. Nat Rev Cancer 9, 338-350, (2009). 30 Minocha, A. & Long, B. H. Inhibition of the DNA Catenation Activity of Type-Ii Topoisomerase by Vp16-213 and Vm26. Biochemical and Biophysical Research Communications 122, 165-170, (1984). 31 Tewey, K. M., Rowe, T. C., Yang, L., Halligan, B. D. & Liu, L. F. Adriamycin-induced DNA damage mediated by mammalian DNA topoisomerase II. Science 226, 466-468, (1984). 32 Marshall, B., Darkin, S. & Ralph, R. K. Evidence That Mamsa Induces Topoisomerase Action. Febs Letters 161, 75-78, (1983). 33 Liu, L. F. DNA Topoisomerase Poisons as Antitumor Drugs. Annual Review of Biochemistry 58, 351-375, (1989). 34 Rao, V. A. et al. The Iron Chelator Dp44mT Causes DNA Damage and Selective Inhibition of Topoisomerase II alpha in Breast Cancer Cells. Cancer Research 69, 948-957, (2009). 35 Downes, C. S. et al. A Topoisomerase Ii-Dependent G2 Cycle Checkpoint in Mammalian-Cells. Nature 372, 467-470, (1994). 36 Clifford, B., Beljin, M., Stark, G. R. & Taylor, W. R. G2 arrest in response to topoisomerase II inhibitors: the role of p53. Cancer Research 63, 4074-4081, (2003). 37 Aubelsadron, G. & Londosgagliardi, D. Daunorubicin and Doxorubicin, Anthracycline Antibiotics, a Physicochemical and Biological Review. Biochimie 66, 333-352, (1984). 38 Fornari, F. A., Randolph, J. K., Yalowich, J. C., Ritke, M. K. & Gewirtz, D. A. Interference by Doxorubicin with DNA Unwinding in Mcf-7 Breast-Tumor Cells. Molecular Pharmacology 45, 649-656, (1994). 39 Keizer, H. G., Pinedo, H. M., Schuurhuis, G. J. & Joenje, H. Doxorubicin (Adriamycin) - a Critical-Review of Free Radical-Dependent Mechanisms of Cytotoxicity. Pharmacology & Therapeutics 47, 219-231, (1990). 40 Singal, P. K. & Iliskovic, N. Doxorubicin-induced cardiomyopathy. New England Journal of Medicine 339, 900-905, (1998). 41 Froelichammon, S. J. & Osheroff, N. Topoisomerase Poisons - Harnessing the Dark Side of Enzyme Mechanism. Journal of Biological Chemistry 270, 21429-21432, (1995). 42 Murray, A. W. Creative Blocks - Cell-Cycle Checkpoints and Feedback Controls. Nature 359, 599-604, (1992). 43 Nakayama, K. I. & Nakayama, K. Ubiquitin ligases: cell-cycle control and cancer. Nat Rev Cancer 6, 369-381, (2006). 44 Zimmermann, K. C., Bonzon, C. & Green, D. R. The machinery of programmed cell death. Pharmacol Ther 92, 57-70, (2001). 45 Soldani, C. et al. Poly(ADP-ribose) polymerase cleavage during apoptosis: When and where? Experimental Cell Research 269, 193-201, (2001). 46 Elmore, S. Apoptosis: A review of programmed cell death. Toxicologic Pathology 35, 495-516, (2007). 47 Ferraro, E. & Cecconi, F. Autophagic and apoptotic response to stress signals in mammalian cells. Archives of Biochemistry and Biophysics 462, 210-219, (2007). 48 Maiuri, M. C., Zalckvar, E., Kimchi, A. & Kroemer, G. Self-eating and self-killing: crosstalk between autophagy and apoptosis. Nature Reviews Molecular Cell Biology 8, 741-752, (2007). 49 Tanida, I., Minematsu-Ikeguchi, N., Ueno, T. & Kominami, E. Lysosomal turnover, but not a cellular level, of endogenous LC3 is a marker for autophagy. Autophagy 1, 84-91, (2005). 50 Ohsumi, Y. Molecular dissection of autophagy: Two ubiquitin-like systems. Nature Reviews Molecular Cell Biology 2, 211-216, (2001). 51 Lindsey, R. H., Jr., Bender, R. P. & Osheroff, N. Effects of benzene metabolites on DNA cleavage mediated by human topoisomerase II alpha: 1,4-hydroquinone is a topoisomerase II poison. Chem Res Toxicol 18, 761-770, (2005). 52 Huang, C. P. et al. Anticancer activity of botanical alkyl hydroquinones attributed to topoisomerase II poisoning. Toxicology and Applied Pharmacology 227, 331-338, (2008). 53 Fortune, J. M. & Osheroff, N. Merbarone inhibits the catalytic activity of human topoisomerase IIalpha by blocking DNA cleavage. J Biol Chem 273, 17643-17650, (1998). 54 Subramanian, D., Furbee, C. S. & Muller, M. T. ICE bioassay. Isolating in vivo complexes of enzyme to DNA. Methods Mol Biol 95, 137-147, (2001). 55 Lin, T. Y. et al. A Cysteine-Reactive Alkyl Hydroquinone Modifies Topoisomerase II alpha, Enhances DNA Breakage, and Induces Apoptosis in Cancer Cells. Chemical Research in Toxicology 25, 2340-2351, (2012). 56 Ron, D. & Habener, J. F. CHOP, a novel developmentally regulated nuclear protein that dimerizes with transcription factors C/EBP and LAP and functions as a dominant-negative inhibitor of gene transcription. Genes Dev 6, 439-453, (1992). 57 Daino, K., Ichimura, S. & Nenoi, M. Both the basal transcriptional activity of the GADD45A gene and its enhancement after ionizing irradiation are mediated by AP-1 element. Biochim Biophys Acta 1759, 458-469, (2006). 58 Zhan, Q. Gadd45a, a p53- and BRCA1-regulated stress protein, in cellular response to DNA damage. Mutat Res 569, 133-143, (2005). 59 Chou, T. C. Drug Combination Studies and Their Synergy Quantification Using the Chou-Talalay Method. Cancer Research 70, 440-446, (2010). 60 Chou, T. C. & Talalay, P. Quantitative-Analysis of Dose-Effect Relationships - the Combined Effects of Multiple-Drugs or Enzyme-Inhibitors. Advances in Enzyme Regulation 22, 27-55, (1984). 61 Li, C. H. et al. Ganoderic acid X, a lanostanoid triterpene, inhibits topoisomerases and induces apoptosis of cancer cells. Life Sciences 77, 252-265, (2005). 62 Mizushina, Y. et al. A novel DNA topoisomerase inhibitor: dehydroebriconic acid, one of the lanostane-type triterpene acids from Poria cocos. Cancer Sci 95, 354-360, (2004). 63 Wang, H. et al. Stimulation of topoisomerase II-mediated DNA damage via a mechanism involving protein thiolation. Biochemistry 40, 3316-3323, (2001). 64 Lin, R. K. et al. Dietary isothiocyanate-induced apoptosis via thiol modification of DNA topoisomerase IIalpha. J Biol Chem 286, 33591-33600, (2011). 65 Tapia, E. et al. Curcumin induces Nrf2 nuclear translocation and prevents glomerular hypertension, hyperfiltration, oxidant stress, and the decrease in antioxidant enzymes in 5/6 nephrectomized rats. Oxid Med Cell Longev 2012, 269039, (2012). 66 Su, C. C. et al. Curcumin-induced apoptosis of human colon cancer colo 205 cells through the production of ROS, Ca2+ and the activation of caspase-3. Anticancer Res 26, 4379-4389, (2006). 67 Fujisawa, S., Atsumi, T., Ishihara, M. & Kadoma, Y. Cytotoxicity, ROS-generation activity and radical-scavenging activity of curcumin and related compounds. Anticancer Res 24, 563-569, (2004). 68 Martin-Cordero, C., Lopez-Lazaro, M., Galvez, M. & Ayuso, M. J. Curcumin as a DNA topoisomerase II poison. Journal of Enzyme Inhibition and Medicinal Chemistry 18, 505-509, (2003). 69 Wilson, W. R., Harris, N. M. & Ferguson, L. R. Comparison of the mutagenic and clastogenic activity of amsacrine and other DNA-intercalating drugs in cultured V79 Chinese hamster cells. Cancer Research 44, 4420-4431, (1984). 70 Dillehay, L. E., Jacobson-Kram, D. & Williams, J. R. DNA topoisomerases and models of sister-chromatid exchange. Mutat Res 215, 15-23, (1989). 71 Pommier, Y., Zwelling, L. A., Kao-Shan, C. S., Whang-Peng, J. & Bradley, M. O. Correlations between intercalator-induced DNA strand breaks and sister chromatid exchanges, mutations, and cytotoxicity in Chinese hamster cells. Cancer Research 45, 3143-3149, (1985). 72 Raina, K. & Agarwal, R. Combinatorial strategies for cancer eradication by silibinin and cytotoxic agents: efficacy and mechanisms. Acta Pharmacol Sin 28, 1466-1475, (2007). | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/62311 | - |
dc.description.abstract | 靈芝是傳統中草藥之一,先前研究發現,靈芝三萜類T-612是一個具有抗癌活性的小分子,但詳細作用機制尚未清楚。拓樸異構酶II (Topoisomerase, Topo II)為解開DNA超螺旋結構的酵素,在DNA複製、轉錄及基因重組時扮演重要角色,許多化療藥物能夠藉由抑制Topo II而達到抑制癌細胞生長的效果。乳癌是全球女性罹患率極高的癌症,在臨床上,雌性素、黃體素受體以及HER2受體均為陰性的三陰性乳癌 (triple negative breast cancer, TNBC)為較惡性、治療效果有限及復發率高的乳癌亞型,在臨床上只能使用化療藥物治療,因此發展治療TNBC的藥物是乳癌治療上重要的課題之一。本篇研究目的是探討T-612抑制Topo II活性的機制與T-612對於TNBC細胞株MDA-MB-231之抗癌應用。In vitro試管實驗結果顯示,T-612能有效抑制Topo II活性 (IC50: 1.2 μM),且抑制的方式為不可逆的,另外,將T-612與Topo II預先反應之後發現,Topo II的活性更顯著受到抑制,推斷T-612能透過直接與Topo II結合而抑制Topo II活性;在in vitro 及in vivo的系統中,T-612皆能造成DNA-Topo II complex累積。T-612處理能夠造成細胞中DNA斷裂、DNA損傷相關基因表現量上升、細胞週期停滯於G2/M時期與細胞凋亡發生,顯示T-612不論是在in vitro或是in vivo實驗,皆有Topo II poison的特性。此外,T-612處理也可造成細胞內活性氧化物 (ROS)大量累積、引發細胞自噬。與乳癌常見化療藥物Doxorubicin合併處理後,T-612可增強Doxorubicin所造成的細胞毒性,細胞中DNA-Topo II complex與ROS累積量增加、細胞週期停滯現象更加顯著,並誘發細胞凋亡。綜合以上結果得知,T-612造成DNA-Topo II complex累積,為一個Topo II poison的藥物;與Doxorubicin合併使用,能藉由增加DNA斷裂與細胞週期停滯,進而造成細胞凋亡,顯示T-612能夠有效增強Doxorubicin的抗癌效果,具有作為Doxorubucin治療TNBC佐劑之潛力。 | zh_TW |
dc.description.abstract | Ganoderma (Lingzhi) has been a notable Chinese traditional medicinal herb for centuries. T-612, a triterpene from Ganoderma tsugae, has been previously found to possess anticancer activity, but the mechanism remains unclear. Breast cancer is the most life threatening malignant disease in women worldwide. In clinical, the breast cancer cells which do not overexpress estrogen receptor, progesterone receptor, and HER-2 genes, called triple negative breast cancer (TNBC) subtype, should be treated by chemotherapy. In this study, we studied anticancer activity of T-612 in MDA-MB-231 cells (a TNBC cell line), and characterized its inhibitory activity on topoisomerase II (Topo II), that is an enzyme essential for DNA replication and transcription, and especially crucial for cancer cell proliferation. T-612 was found to irreversibly inhibit Topo II activity with an IC50 of 1.2 μM. Both in vitro and in vivo, T-612 treatment caused Topo II poison, accumulation of the Topo II-DNA complex. In addition, T-612 caused DNA breaks, G2/M arrest, DNA damage-related gene expression and apoptosis, which are responses caused by Topo II poison. T-612 also caused reactive oxygen species (ROS) accumulation and induced autophagy. We further investigated whether T-612 is able to enhance the efficacy of Doxorubicin (another Topo II inhibitor) through further Topo II inhibition. The combination of T-612 and Doxorubicin enhanced DNA-Topo II complex formation and ROS accumulation in MDA-MB-231 cells. This combined treatment displayed synergistic cytotoxicity through higher level of G2/M arrest and apoptosis compared to Doxorubicin alone. The synergistic effect of T-612 and Doxorubicin in MDA-MB-231 cells indicated that the T-612 might be a potential anticancer adjuvant for treatment of TNBC. | en |
dc.description.provenance | Made available in DSpace on 2021-06-16T13:40:18Z (GMT). No. of bitstreams: 1 ntu-102-R00424017-1.pdf: 4559551 bytes, checksum: e57c3654b4f2c4a0bbd45a2dc7d1c963 (MD5) Previous issue date: 2013 | en |
dc.description.tableofcontents | 中文摘要 I
Abstract II 英文縮寫表 III 示意圖目錄 VI 圖目錄 VI 一、研究背景 1 1.1 靈芝 (Ganoderma) 1 1.2 乳癌 1 1.3 拓樸異構酶 (Topoisomerase, Topo) 5 1.4 Topo II的作用 6 1.5 Topo II抑制劑 6 1.6 多柔比星 (Doxorubicin, Doxo) 7 1.7 細胞週期 (Cell cycle) 8 1.8 計畫性細胞死亡 (programmed cell death) 9 1.9 研究目的 11 二、材料與方法 12 2.1 試管實驗分析Topo II解螺旋的活性 12 2.2 兩階段Topo II活性測定 12 2.3 藥物嵌入DNA測試 13 2.4 試管實驗分析DNA 斷裂的形成 13 2.5 細胞培養及藥物處理條件 13 2.6 細胞存活率測定 (ACP assay) 14 2.7 Immunodetection of in vivo complexes of enzyme to DNA (ICE bioassay) 14 2.8 免疫螢光染色 15 2.9 流式細胞儀測定 16 2.10 反轉錄酵素反應及即時聚合酶連鎖反應 17 2.11 蛋白質萃取及西方墨點法 18 2.12 統計分析 18 三、實驗結果 19 3.1 T-612抑制 Topo II活性且為Topo II poison的藥物 19 3.2 T-612造成Topo II導致的DNA斷裂並促進DNA受損相關基因表現 21 3.3 T-612處理造成細胞週期停滯於G2/M時期,並增加sub-G1比例 23 3.4 T-612處理引發細胞內ROS累積 23 3.5 T-612處理引發細胞自噬及細胞凋亡 23 3.6 T-612與Doxorubicin合併處理於MDA-MB-231細胞為協同作用,相較於單獨處理有更顯著的毒殺效果。 24 3.7 合併使用T-612及Doxorubicin造成細胞週期停滯於G2/M時期並造成sub-G1比例增加 25 3.8 合併T-612與Doxorubicin造成細胞凋亡情形增加 25 3.9 合併T-612與Doxorubicin造成細胞ROS累積量上升,但氧化壓力並非造成細胞死亡的主要原因 26 四、討論 27 五、圖 30 參考文獻 50 藥品清單 58 | |
dc.language.iso | zh-TW | |
dc.title | 探討靈芝三萜類T-612之抗癌分子機制 | zh_TW |
dc.title | Anticancer Molecular Mechanism of Ganoderma Triterpene T-612 | en |
dc.type | Thesis | |
dc.date.schoolyear | 101-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 胡忠怡(Chung-Yi Hu),張雅雯(Ya-Wen Chang),楊雅倩(Ya-Chien Yang) | |
dc.subject.keyword | 靈芝三?類,拓樸異構?II,三陰性乳癌,Doxorubicin,抗癌佐劑, | zh_TW |
dc.subject.keyword | Ganoderma triterpene,Topoisomerase II,triple negative breast cancer,Doxorubicin,anticancer adjuvant, | en |
dc.relation.page | 61 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2013-07-15 | |
dc.contributor.author-college | 醫學院 | zh_TW |
dc.contributor.author-dept | 醫學檢驗暨生物技術學研究所 | zh_TW |
顯示於系所單位: | 醫學檢驗暨生物技術學系 |
文件中的檔案:
檔案 | 大小 | 格式 | |
---|---|---|---|
ntu-102-1.pdf 目前未授權公開取用 | 4.45 MB | Adobe PDF |
系統中的文件,除了特別指名其著作權條款之外,均受到著作權保護,並且保留所有的權利。